RU2804665C2 - Compositions and methods of gene editing - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to a fused protein containing in the direction from the N-terminus to the C-terminus a DNA methyltransferase domain, an enzyme nuclease-deficient RNA-directed DNA endonuclease and a domain associated with Krüppel box or Krüppel box related domain, the enzyme nuclease-deficient RNA-directed DNA endonuclease and the domain of DNA methyltransferase, as well as the complex containing it. Also a nucleic acid molecule encoding the above fusion protein, as well as a vector and a cell containing it are disclosed.

EFFECT: invention is effective for silencing a target nucleic acid sequence in a cell.

60 cl, 13 dwg, 2 tbl, 12 ex

Description

Cross-reference to related applications

[0001]This The application claims priority to U.S. Patent Application No. 62/660023, filed April 19, 2018, the disclosure of which is incorporated herein by reference in its entirety.

STATEMENT REGARDING RIGHTS TO INVENTIONS RESULTING FROM FEDERALLY SPONSORED RESEARCH WORK

[0002] This invention is supported by the United States Government under Grant No. HR0011-17-2-0043 awarded by the Department of Defense, Defense Advanced Research Projects Agency, and under Grant No. R01 DA036858 awarded by the National Institutes of Health. The US Government has certain rights to this invention.

BACKGROUND OF THE ART

[0003] Although genome editing is considered a promising therapeutic approach for treating diseases, it carries inherent risks due to potential genotoxicity associated with double-strand breaks (DSBs). In addition, genome editing is often associated with an all-or-nothing effect on the target gene (i.e., it causes a complete knockout). In contrast, targeted epigenome engineering does not carry the risk of DSB-induced genotoxicity; in addition, it makes it possible to create a more differentiated effect on gene expression and, thus, provide functioning from complete silencing to a less pronounced effect.

[0004] This document provides solutions to these and other needs in the art.

BRIEF DESCRIPTION OF THE INVENTION

[0005] In one aspect of the present invention, there is provided a fusion protein comprising a nuclease-deficient RNA-guided DNA endonuclease enzyme box-linked domain (KRAB domain), and DNA methyltransferase domain. In one aspect of the present invention, a fusion protein of any of the sequences of SEQ ID NO: 1-15 is provided.

[0006] In one aspect of the present invention, there is provided a nucleic acid sequence encoding a fusion protein as described herein, inclusive of embodiments and aspects thereof.

[0007] In one aspect of the present invention, there is provided a complex including a fusion protein as described herein, inclusive of embodiments and aspects thereof, and a polynucleotide including (1) a DNA targeting sequence that is complementary to the target polynucleotide sequence and (2) a binding sequence for a nuclease-deficient RNA-directed DNA endonuclease enzyme, wherein the nuclease-deficient RNA-directed DNA endonuclease enzyme is linked to the polynucleotide through the binding sequence.

[0008] In one aspect of the present invention, there is provided a vector including a nucleic acid sequence of a fusion protein as described herein, inclusive of embodiments and aspects thereof.

[0009] In one aspect of the present invention, there is provided a cell including a fusion protein as described herein, inclusive of embodiments and aspects thereof, a nucleic acid as described herein, inclusive of embodiments and aspects thereof, a complex, as described herein, inclusive of embodiments and aspects thereof, or a vector, as described herein, inclusive of embodiments and aspects thereof.

[0010] In one aspect of the present invention, there is provided a method for silencing a target nucleic acid sequence in a cell, including delivering a first polynucleotide encoding a fusion protein as described herein, inclusive of embodiments and aspects thereof, into a cell containing the target nucleic acid, and delivery into the cell of a second polynucleotide that includes (i) a DNA targeting sequence that is complementary to the target nucleic acid sequence, and (ii) a binding sequence for the enzyme nuclease-deficient RNA-guided DNA endonuclease. Without intending to be bound by any theory, the fusion protein is believed to cause silencing of a target nucleic acid sequence in a cell by methylating the chromatin containing the target nucleic acid sequence and/or by introducing repressive chromatin markers into the chromatin containing the target nucleic acid sequence . Thus, in aspects of the present invention, the fusion protein causes silencing of a target nucleic acid sequence in a cell by methylating chromatin containing the target nucleic acid sequence and/or by introducing repressive chromatin markers into chromatin containing the target nucleic acid sequence.

[0011] In one aspect of the present invention, there is provided a method for silencing a target nucleic acid sequence in a cell, including delivering a complex as described herein, inclusive of embodiments and aspects thereof, into a cell containing the target nucleic acid, wherein said complex causes silencing sequences of a target nucleic acid in a cell. Without intending to be bound by any theory, the complex is believed to cause silencing of a target nucleic acid sequence in a cell by methylating the chromatin containing the target nucleic acid sequence and/or by introducing repressive chromatin markers into the chromatin containing the target nucleic acid sequence. Thus, in aspects of the present invention, the complex causes silencing of a target nucleic acid sequence in a cell by methylating the chromatin containing the target nucleic acid sequence and/or by introducing repressive chromatin markers into the chromatin containing the target nucleic acid sequence.

BRIEF DESCRIPTION OF GRAPHIC MATERIALS

[0012] In FIG. 1A-1F describe all-in-one protein design for long-term gene silencing. In FIG. 1A is a schematic representation of an all-in-one protein (SEQ ID NO: 1) of the present invention that contains a KRAB domain fused to the -N terminus of dCas9 (SEQ ID NO: 23) separated by a GGSGGGS linker (SEQ ID NO : 17), and Dnmt3A - Dnmt3L at the C-terminus of dCas9 (separated using the linker EASGSGRASPGIPGSTR (SEQ ID NO: 19)). In FIG. Figure 1B shows schematic representations of dCas9-fused epigenetic modulators tested for permanent gene silencing. The dCas9 - KRAB protein is adapted from Gilbert et al., Cell 2013 for use with CRISPR interference (CRISPRi) technology. The dCas9 - Dnmt3A - Dnmt3L fusion protein is adapted from Stepper et al., Nucleic Acids Research, 2016. We have designed a new all-in-one protein that combines the KRAB domain (SEQ ID NO: 16), dCas9 (D10A, H208A) , Dnmt3A - Dnmt3L (SEQ ID NO: 33; where Dnmt3A is SEQ ID NO: 26 and Dnmt3L is SEQ ID NO: 28) into one polypeptide. In FIG. Figure 1C shows schematic representations of a methylation-sensitive GFP reporter (adapted from Stelzer et al., Cell 2015) that was used to evaluate long-term silencing induced by the all-in-one protein. In FIG. 1D-1E shows a diagram and results of an experimental hit-and-run mutagenesis workflow in HEK293T cells expressing the GFP reporter shown in FIG. 3. In FIG. Figure 1D shows that plasmids were cotransfected into cells, one of which encoded the hit-and-run protein, and the other plasmid encoded the egRNA. In FIG. Figure 1E shows the results of a hit-and-run mutagenesis assay, sorting cells that were cotransfected with both the all-in-one plasmid and the segRNA plasmid. In FIG. Figure 1F shows that the results of GFP reporter silencing depend on the egRNA sequence.

[0013] In FIG. 2A-2F describe long-term silencing of endogenous genes. In FIG. 2A-2C show representative flow cytometry data obtained 22 days after transfection followed by targeting genes (CD29, CD81, CD151) for long-term silencing using an all-in-one protein. Quadrant IV represents cells that have turned off the corresponding gene, as indicated by the percentage of cells with the gene turned off (i.e., 45%, 66%, and 53%, respectively). In FIG. Figure 2D shows a quantitative assessment of the silencing of the CD29, CD81 and CD151 genes using three different egRNAs. In FIG. Figure 2E quantifies the silencing of two or three genes simultaneously to show that an all-in-one protein can be multiplexed by co-delivering egRNA molecules targeting different genes. In FIG. 2F is a graph that describes a time point taken 9 months after transfection of the all-in-one protein and segRNA targeting the CLTA gene, which indicates that the majority of cells have stably silenced the CLTA gene.

[0014] In FIG. 3A-3I describe long-term silencing of endogenous genes. In FIG. 3A-3C show that the harvested cells lost expression of CD29 (Figure 3A), CD81 (Figure 3B) and CD151 (Figure 3C) thirty-six days post-transfection, as determined by their RNA expression profiles. Fig. 3D-3F are volcano plots showing that the target genes CD29 (Figure 3D), CD81 (Figure 3E) and CD151 (Figure 3F) are the only significant gene knockdown for each experiment, indicating the high specificity of gene silencing. In FIG. Figures 3G-3I show quantification of CD151 (Figure 3G), CD81 (Figure 3H), and CD29 (Figure 3I) transcript levels, demonstrating >96% knockdown of each target gene.

[0015] In FIG. 4A-4H describe long-term gene silencing in various mammalian cell lines. Fig. 4A - 4F are flow cytometry plots showing the expression of blue fluorescent protein (BFP) (which is fused to an all-in-one protein) in HeLa (cervical cells) (Figure 4A), U2OS (bone cells tissue) (Figure 4B) and in human induced pluripotent stem cells (iPSCs) (Figure 4C). Fig. 4D-4F represent untransfected controls for Fig. 4A-4C, respectively. In FIG. 4G shows that stable silencing of endogenous genes was achieved in HeLa and U2OS cells, measured 18 days after transfection with the all-in-one protein. In FIG. 4A-4F, the x-axis is SFB (all-in-one protein fusion) and the y-axis is mCherry. In FIG. 4H shows that gene silencing was detected 14 days post-transfection by qPCR in mouse AML12 hepatocyte cell lines when targeting Pcsk9, Npc1, Spcs1 and Cd81.

[0016] In FIG. 5 shows diagrams of the fusion proteins p76, p90 - p102 and p112, which correspond to the sequences of SEQ ID NO: 1-15, respectively.

[0017]In FIG. 6A-6E steps for gene silencing of all-in-one protein variants are described. In FIG. 6A-6B show the results of gene silencing 18 days after transfection of fusion proteins of SEQ ID NOs: 1-15 transfected into HEK293T cells to specifically silence the CLTA gene. The dCas9 - KRAB and dCas9 - Dnmt3A - Dnmt3L formats demonstrated short-term and lower long-term silencing efficiency. In FIG. 6C-6D show a comparison of SEQ ID NO: 1 (p76) and SEQ ID NO: 15 (p112) in the silencing of the endogenous HIST2H2BE (H2B) gene (Fig. 6C) and the synthetic reporter gene Snrpn-GFP (Fig. 6D), stable expressed in HEK293T cells. In FIG. 6E shows a graph of protein expression (dashed lines) of p76 and p112 over a 50-day period to silence the HIST2H2BE (H2B) gene. Protein levels were measured using a flow cytometry method to detect SBF, which is co-expressed with the all-in-one protein.

[0018] In FIG. 7A-7B show Western blots of all-in-one protein variants. In FIG. 7A shows Western blot analysis of all-in-one protein variants p76 and p90 - p102 using the anti- Steptococcus pyogenes Cas9 antibody. The top band represents the full-length protein, and the smaller bands represent the proteolysis products of the all-in-one proteins. In FIG. 7B shows a Western blot analysis of all-in-one protein variants to detect free Dnmt3A that is cleaved from the fusion protein.

[0019] In FIG. 8A-8E describe a pooled screen to identify optimal egRNAs. In FIG. Figure 8A presents a scheme for a pooled screen to identify optimal egRNAs that lead to long-term gene silencing. In FIG. 8B-8E are flow cytometry histograms showing the percentage of cells undergoing gene silencing four weeks after transfection. Four HEK293T cell lines were used, each with a different GFP-tagged gene, including CLTA (Figure 8B), VIM (Figure 8C), HIST2H2BE (H2B) (Figure 8D), and RAB11A (Figure 8E).

[0020] In FIG. 9A-9D show maps of egRNA functionality at the transcription start site of the target gene, including CLTA (Fig. 9A), H2B (Fig. 9B), RAB11 (Fig. 9C), and VIM (Fig. 9D). The transcription start site (TSS) and CpG island are annotated above each graph. Each dot represents one egRNA, and its effectiveness in long-term gene silencing is displayed as a change in log2-fold egRNA content. The degree of nucleosome occupancy (lower graph) is plotted against the signal from MNase (micrococcal nuclease, English “MNase”).

[0021]In FIG. 10A-10E Functional egRNAs for long-term gene silencing have been described. In FIG. 10A shows a pooled screening workflow in HEK293T cells to determine optimal segRNA targeting positions for an all-in-one protein, adapted from a previous high-density ricin bioarray screen in K562 cells to identify optimal segRNAs for dCas9 - KRAB (Gilbert, Horlbeck et al. , Cell 2014). In FIG. 10B-10E are representative plots showing growth phenotypes for four genes, including ARL1 (Figure 10B), EIF6 (Figure 10C), SMC3 (Figure 10D), HEATR1 (Figure 10E), from existing data sets for dCas9 - KRAB/ CRISPRi in K562 cells (Gilbert, Horlbeck et al., 2014), and with the all-in-one protein (lower graph). Each dot represents an egRNA. The CNT and annotated CpG island are shown for each gene.

[0022] In FIG. 11A-11B are a comparison of the growth and nucleosome positioning phenotypes (as signaled by MNase) for VPS53 (Fig. 11A) and VPS54 (Fig. 11B), which shows the location of functional egRNAs in regions lacking nucleosomes.

[0023] In FIG. 12A-12C show all-in-one protein delivery via mRNA expression. In FIG. 12A shows in vitro transcription of two all-in-one protein variants (p102 and p112) and also shows full-length synthesis of each construct. In FIG. 12B is a flow cytometry graph showing the expression of p102 and p112 one day after transfection of the mRNA into HEK293T cells. In FIG. Figure 12C shows the dynamics of endogenous CLTA gene silencing in HEK293T cells after transfection with mRNA expressing all-in-one protein variants p102 and p112.

[0024] In FIG. 13A-13G describe controlled expression of an all-in-one protein using doxycycline induction. In FIG. 13A are flow cytometry plots showing the induced expression of the all-in-one protein by addition of doxycycline in K562 cells that stably encode the all-in-one protein under the influence of a doxycycline-inducible promoter. The dotted line represents the baseline median fluorescence of SPB without doxycycline administration. In FIG. 13B shows Western blots of cells to detect all-in-one protein expression before and after doxycycline treatment. In FIG. 13C-13F show flow cytometry plots for knockdown of the CD81 (FIGS. 13C-13D) and CD151 (FIGS. 13E-13F) genes 14 days after doxycycline treatment of K562 cells. In FIG. 13G presents a quantitative assessment of the knockdown of the CD81 and CD151 genes 14 days after treatment with doxycycline or without treatment with doxycycline.

DETAILED DESCRIPTION OF THE INVENTION

[0025] The technology described herein allows, among other things, permanent gene silencing in mammalian cells without the formation of DNA double-strand breaks in the host genome. In embodiments of the present invention, the core component is a single polypeptide chain consisting of catalytically inactive Cas9 (dCas9) fused to Dnmt3A, Dnmt3L, and a KRAB domain (herein referred to as the “all-in-one protein”). The fusion protein provided herein can be targeted to a specific site in the mammalian genome using a single guide RNA (sgRNA), and can result in DNA methylation and/or introduce repressive chromatin markers at that site. In embodiments of the present invention, the result is gene silencing, which is inherited through subsequent cell divisions. In embodiments of the present invention, the fusion protein provided herein (and egRNA) are expressed only transiently, bypassing the use of viral delivery methods to induce permanent silencing.

[0026] In embodiments of the present invention, the fusion proteins provided herein provide robust long-term or permanent silencing of endogenous gene expression through editing of the epigenome rather than the genome. It is possible to target both alleles of a gene or to selectively target one pathogenic allele. In embodiments of the present invention, an advantage of the fusion protein provided herein is that epigenetic editing is reversible and therefore inherently safer than genome editing. Thus, in embodiments of the present invention, the fusion protein provided herein is useful for prophylactic purposes. For example, gene silencing may provide acute protection against infection/biological toxin, and then silencing may be reversed once there is no risk of infection or toxicity. Thus, in embodiments of the present invention, the fusion protein provided herein has beneficial properties in a situation in which a virus or toxin enters a cell through interaction with a protein that is required for long-term organ function or homeostasis. In embodiments of the present invention, the fusion protein provided herein is useful in genome editing-based therapeutics.

[0027] In embodiments of the present invention, permanent gene silencing in mammalian cells can be achieved using two components: a single polypeptide chain consisting of dCas9 fused to three epigenetic modulators, and a single guide RNA that directs the protein to a specific site in the host genome . In embodiments of the present invention, the components are expressed only transiently in the host cell, thereby reducing toxicity and off-target effects.

[0028] In embodiments of the present invention, the fusion protein provided herein does not cause DNA breaks in the host cell to ensure permanent gene silencing. In embodiments of the present invention, the epigenetic marks that are added to the genomic site of interest are reversible, allowing the removal of any off-target events that may occur.

[0029] Definitions

[0030] Although various embodiments and aspects of the present invention are shown and described herein, those skilled in the art will appreciate that such embodiments and aspects are presented by way of example only. Numerous variations, changes and substitutions will now be apparent to those skilled in the art without departing from the present invention. It should be understood that various alternatives to the embodiments of the invention described herein may be used in the practice of the invention.

[0031] The section headings used herein are for organizational purposes only and should not be construed as limiting the described subject matter of the present invention. All documents or portions of documents cited in this application, including, but not limited to, patents, patent applications, articles, books, manuals and scientific treatises, are hereby incorporated by reference in their entirety for all purposes.

[0032] Unless otherwise specified, all technical and scientific terms used herein have the meaning commonly understood by one skilled in the art to which this invention belongs. The following references provide one skilled in the art with a general definition of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology ( ^2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, ^5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used in this document, the following terms have the meanings ascribed to them unless otherwise noted.

[0033] The use of singular or plural in this specification and in the following claims follows the traditional approach in patents to mean “at least one”, unless in a particular instance it is clear from the context that the term is intended in that particular instance to denoting one thing and one thing only. Likewise, the term “comprising” is non-limiting and does not exclude additional elements, functions, components, etc. Literary sources cited herein are expressly incorporated herein by reference in their entirety unless otherwise noted.

[0034] The terms “comprise”, “include” and “have”, as well as their derivatives, are used interchangeably in the context of this document as broad, non-limiting terms. For example, when using the terms “containing,” “including,” or “having,” it is implied that whatever element is contained, included, or had is not the only element covered by the subject of the sentence containing the corresponding verb above.

[0035] The term "nucleic acid" refers to nucleotides (eg, deoxyribonucleotides or ribonucleotides) and polymers thereof in single-stranded, double-stranded or multistranded form, or their complements. The terms "polynucleotide", "oligonucleotide", "oligo" and the like refer in the ordinary and accepted sense to a linear sequence of nucleotides. The term "nucleotide" refers in the ordinary and accepted sense to a single unit of polynucleotide, that is, a monomer. Nucleotides may be ribonucleotides, deoxyribonucleotides, or modified versions thereof. Examples of polynucleotides provided herein include single- and double-stranded DNA, single- and double-stranded RNA, and hybrid molecules containing mixtures of single- and double-stranded DNA and RNA. Examples of nucleic acids, such as polynucleotides, provided herein include, but are not limited to, any type of RNA, such as mRNA, miRNA, microRNA, segRNA, and guide RNA, and any type of DNA, such as genomic DNA, plasmid DNA, and minicircular DNA. , and any fragments thereof. In aspects of the present invention, the nucleic acid is messenger RNA. In some aspects of the present invention, the messenger RNA is a messenger ribonucleoprotein (RNP). The term "duplex" as used herein in the context of polynucleotides refers, in the usual and accepted sense, to double-strandedness. Nucleic acids can be linear or branched. For example, nucleic acids may be a linear chain of nucleotides, or nucleic acids may be branched, such that the nucleic acids contain one or more nucleotide arms or branches. Optionally, branched nucleic acids are branched multiple times to form higher order structures such as dendrimers and the like.

[0036] As used herein, the terms “nucleic acid,” “nucleic acid molecule,” “nucleic acid oligomer,” “oligonucleotide,” “nucleic acid sequence,” “nucleic acid fragment,” and “polynucleotide” are used interchangeably and are intended to include , but not limited to, a polymeric form of nucleotides covalently linked together, which may be of varying lengths, either deoxyribonucleotides or ribonucleotides, or analogs, derivatives or modifications thereof. Different polynucleotides may have different three-dimensional structures and may perform a variety of functions, known or unknown. Non-limiting examples of polynucleotides include a gene, gene fragment, exon, intron, intergenic DNA (including, without limitation, heterochromatic DNA), messenger RNA (mRNA), transfer RNA, ribosomal RNA, ribozyme, cDNA, recombinant polynucleotide, branched-chain polynucleotide, plasmid , vector, isolated DNA sequence, isolated RNA sequence, sgRNA, guide RNA, nucleic acid probe and primer. The polynucleotides used in the methods of this invention may contain natural nucleic acid sequences and variants thereof, artificial nucleic acid sequences, or a combination of such sequences.

[0037] A polynucleotide typically consists of a specific sequence of four nucleotide bases: adenine (A); cytosine (C); guanine (G); and thymine (T) (uracil (U) instead of thymine (T) when the polynucleotide is RNA). Thus, the term "polynucleotide sequence" is an alphabetical representation of a polynucleotide molecule; alternatively, the term may apply to the polynucleotide molecule itself. Such an alphabetical representation can be entered into databases on a computer having a central processing unit and have bioinformatic applications such as use in functional genomics and homology searches. Polynucleotides may optionally include one or more non-standard nucleotides, nucleotide analogs and/or modified nucleotides.

[0038] Nucleic acids, including, for example, nucleic acids with a phosphorothioate backbone, may include one or more reactive moieties. As used herein, the term “reactive moiety” includes any group capable of reacting with another molecule, such as a nucleic acid or polypeptide, through covalent, non-covalent or other interactions. By way of example, a nucleic acid may include an amino acid-reactive moiety that reacts with an amino acid on a protein or polypeptide through a covalent, non-covalent, or other interaction.

[0039] The terms also include nucleic acids containing known nucleotide analogues or modified backbone residues or linkages that are synthetic, naturally occurring and non-naturally occurring, that have the same binding properties as the reference nucleic acid, and that are metabolized similar to reference nucleotides. Examples of such analogues include, but are not limited to, phosphodiester derivatives, including, for example , phosphoramidate, phosphorodiamidate, phosphorothioate (also known as phosphorothioate containing, through a double bond, sulfur that has replaced the oxygen in the phosphate), phosphorodithioate, phosphonocarboxylic acids, phosphonocarboxylates, phosphonoacetic acids , phosphonoformic acid, methylphosphonate, borophosphonate or O-methylphosphoramidite linkages (see Eckstein, Oligonucleotides and Analogues: A Practical Approach, Oxford University Press), and modifications of nucleotide bases such as 5-methylcytidine or pseudouridine; and peptide nucleic acid backbones and linkages. Other nucleic acid analogues include positive-backbone nucleic acids; nonionic backbones, modified sugars, and non-ribose backbones (e.g., phosphorodiamidate morpholine oligonucleotides or locked nucleic acids (LNAs) as known in the art), including those described in US Pat. No. 5,235,033 and US Pat. No. 5,034,506 , and in Chapters 6 and 7 of ASC Symposium Series 580, Carbohydrate Modifications in Antisense Research, Sanghui & Cook, eds. Nucleic acids containing one or more carbocyclic sugars are also included in one definition of nucleic acids. Modifications to the ribose phosphate backbone can be made for a variety of reasons, such as to improve the stability and half-life of such molecules in a physiological environment or as probes on a biochip. Mixtures of naturally occurring nucleic acids and analogues can be prepared; alternatively, mixtures of various nucleic acid analogs and mixtures of naturally occurring nucleic acids and analogs can be prepared. In aspects of the present invention, the internucleotide linkages in the DNA are phosphodiester linkages, derivatives of phosphodiester linkages, or a combination thereof.

[0040] Nucleic acids may include non-specific sequences. As used herein, the term “non-specific sequence” refers to a nucleic acid sequence that contains a series of residues not intended to be complementary to, or only partially complementary to, any other nucleic acid sequence. By way of example, a nonspecific nucleic acid sequence is a sequence of nucleic acid residues that does not function as an inhibitory nucleic acid upon contact with a cell or organism.

[0041] The term “complementary” or “complementarity” refers to the ability of a nucleic acid to form hydrogen bond(s) of either conventional Watson-Crick type or other non-traditional types with another nucleic acid sequence. For example, the sequence A - G - T is complementary to the sequence T - C - A. Percent complementarity indicates the percentage of residues in a nucleic acid molecule that can form hydrogen bonds (e.g., Watson-Crick base pairing) with a second nucleic acid sequence (e.g., 5 , 6, 7, 8, 9, 10 out of 10 are 50%, 60%, 70%, 80%, 90% and 100% complementary, respectively). The term “perfectly complementary” means that all contiguous residues of a nucleic acid sequence will hydrogen bond to the same number of contiguous residues in a second nucleic acid sequence. As used herein, the term “substantially complementary” refers to a degree of complementarity that is at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 97%, 98% , 99% or 100% in the area of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40 , 45, 50 or more nucleotides, or refers to two nucleic acids that hybridize under stringent conditions (ie, stringent hybridization conditions).

[0042] The phrase “stringent hybridization conditions” refers to the conditions under which a probe will hybridize to its target subsequence, typically in a complex mixture of nucleic acids, but not to other sequences. The stringent conditions depend on the sequence and will be different in different circumstances. Longer sequences hybridize specifically at higher temperatures. A detailed guide to nucleic acid hybridization can be found in Tijssen, Techniques in Biochemistry and Molecular Biology - Hybridization with Nucleic Probes, “Overview of principles of hybridization and the strategy of nucleic acid assays” (1993). Typically, stringent conditions are selected that are approximately 5-10°C lower than the melting point temperature (Tm) for a particular sequence at a particular ionic strength and pH. Tm is the temperature (at a certain ionic strength, pH and nucleic acid concentration) at which 50% of the probes complementary to the target hybridize with the target sequence at equilibrium (since the target sequences are present in excess, then at Tm 50% probes are occupied at equilibrium). Harsh conditions can also be achieved by adding destabilizing agents such as formamide. For selective or specific hybridization, the positive signal must be at least twice the background, preferably 10 times the background hybridization. Examples of stringent hybridization conditions may be: 50% formamide, 5x SSC and 1% SDS, incubated at 42°C, or 5x SSR, 1% SDS, incubated at 65°C C, with a wash in 0.2x SCP and 0.1% SDS at 65°C.

[0043] Nucleic acids that do not hybridize to each other under stringent conditions are still substantially identical if the polypeptides they encode are substantially identical. This occurs, for example, when a copy of a nucleic acid is made using the maximum codon degeneracy allowed by the genetic code. In such cases, the nucleic acids are typically hybridized under moderately stringent hybridization conditions. Examples of “moderately stringent hybridization conditions” include hybridization in a buffer containing 40% formamide, 1 M NaCl, 1% SDS at 37°C and a wash in 1X SCP at 45°C. Positive hybridization is one that is at least twice the background hybridization. One of ordinary skill in the art will readily appreciate that alternative hybridization and washing conditions can be used to provide conditions of similar stringency. Additional guidance on determining hybridization parameters is provided in numerous references, for example, Current Protocols in Molecular Biology, ed. Ausubel, et al., supra.

[0044] The term "gene" means a segment of DNA involved in the production of a protein; it includes the regions preceding and following the coding region (leading and lagging), as well as intermediate sequences (introns) between individual coding segments (exons). Leading and lagging regions, as well as introns, include regulatory elements that are required during gene transcription and translation. In addition, a “protein gene product” is a protein expressed by a particular gene.

[0045] The word "expression" or "expressed" as used herein in relation to a gene means the product of transcription and/or translation of that gene. The level of expression of a DNA molecule in a cell can be determined either based on the amount of corresponding mRNA that is present in the cell or based on the amount of protein encoded by that DNA produced by the cell. The level of expression of non-coding nucleic acid molecules (eg, egRNA) can be determined by standard PCR or Northern blotting methods well known in the art. See Sambrook et al., 1989 Molecular Cloning: A Laboratory Manual, 18.1-18.88.

[0046] The term “transcription regulatory sequence” as used herein refers to a segment of DNA that is capable of increasing or decreasing transcription (eg, expression) of a particular gene in an organism. Non-limiting examples of transcriptional regulatory sequences include promoters, enhancers, and silencers.

[0047] The terms "transcription start site" and "transcription start site" may be used interchangeably to refer, as used herein, to the 5' end of a gene sequence (e.g., DNA sequence) where the RNA polymerase (e.g., DNA-directed RNA polymerase ) begins the synthesis of the RNA transcript. The transcription start site may be the first nucleotide of the transcribed DNA sequence where RNA polymerase begins to synthesize the RNA transcript. One skilled in the art can determine the transcription start site through routine experimentation and analysis, for example, by performing an “interrupted transcription” analysis, or by using definitions in accordance with the FANTOM5 database.

[0048] The term "promoter" as used herein refers to a region of DNA that initiates transcription of a particular gene. Promoters are typically located near the transcription start site of a gene, upstream of the gene, and on the same DNA strand (i.e., position 5' on the sense DNA strand). Promoters can be from about 100 to about 1000 base pairs in length.

[0049] The term "enhancer" as used herein refers to a region of DNA that can be bound by proteins (eg, transcription factors) to increase the likelihood that transcription of a gene will occur. Enhancers can range in length from about 50 to about 1500 base pairs. Enhancers can be located downstream or upstream of the transcription start site they regulate and can be several hundred base pairs away from the transcription start site.

[0050] The term "silencer" as used herein refers to a DNA sequence that is capable of binding transcription regulatory factors known as repressors, thereby negatively affecting the transcription of a gene. DNA silencer sequences can be found at many different positions along the length of DNA, including, but not limited to, upstream of the target gene at which they act by inhibiting gene transcription (eg, causing silencing of gene expression).

[0051] The term “guide RNA” or “gRNA” as used herein refers to any polynucleotide sequence having sufficient complementarity to a target polynucleotide sequence to hybridize to the target sequence and directly bind the CRISPR complex to the target sequence in a sequence-specific manner. In aspects of the present invention, the degree of complementarity between a guide sequence and its corresponding target sequence when optimally aligned using a suitable alignment algorithm is about or greater than about 50%, 60%, 75%, 80%, 85%, 90%, 95%, 97 .5%, 99%, or more.

[0052] In embodiments of the present invention, the polynucleotide (eg, gRNA) is a single-stranded ribonucleic acid. In aspects of the present invention, the polynucleotide (eg, gRNA) is 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 or more nucleic acid residues in length. In aspects of the present invention, the polynucleotide (eg, gRNA) has a length of from 10 to 30 nucleic acid residues. In aspects of the present invention, the polynucleotide (eg, gRNA) is 20 nucleic acid residues in length. In aspects of the invention, the length of the polynucleotide (e.g., gRNA) may be at least 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100 or more nucleic acid residues or sugar residues in length. In aspects of the invention, the polynucleotide (e.g., gRNA) has a length of 5 to 50, 10 to 50, 15 to 50, 20 to 50, 25 to 50, 30 to 50, 35 to 50, 40 to 50, from 45 to 50, from 5 to 75, from 10 to 75, from 15 to 75, 20 to 75, from 25 to 75, from 30 to 75, from 35 to 75, from 40 to 75, from 45 to 75 , from 50 to 75, from 55 to 75, from 60 to 75, from 65 to 75, from 70 to 75, from 5 to 100, from 10 to 100, from 15 to 100, from 20 to 100, from 25 to 100 , from 30 to 100, from 35 to 100, from 40 to 100, from 45 to 100, from 50 to 100, from 55 to 100, from 60 to 100, from 65 to 100, from 70 to 100, 75 to 100, 80 to 100, 85 to 100, 90 to 100, 95 to 100 or more residues. In aspects of the present invention, the polynucleotide (eg, gRNA) has a length of 10 to 15, 10 to 20, 10 to 30, 10 to 40, or 10 to 50 residues.

[0053] The term “amino acid” refers to naturally occurring and synthetic amino acids, as well as amino acid analogs and amino acid mimetics, which act similarly to naturally occurring amino acids. Naturally occurring amino acids are those amino acids that are encoded by the genetic code, as well as those amino acids that are later modified, such as hydroxyproline, γ-carboxyglutamate, and O-phosphoserine. Amino acid analogues refer to compounds that have the same basic chemical structure as naturally occurring amino acids, i.e., an α-carbon bonded to a hydrogen, a carboxyl group, an amino group, and an R group, e.g., homoserine, norleucine, methionine sulfoxide, methionine methylsulfonium . Such analogues have modified R groups (eg, norleucine) or modified peptide backbones, but retain the same basic chemical structure as the naturally occurring amino acid. Amino acid mimetics refer to chemical compounds that have a structure different from the general chemical structure of an amino acid, but that act in a manner similar to that of naturally occurring amino acids. The terms “non-naturally occurring amino acid” and “non-naturally occurring amino acid” refer to amino acid analogues, synthetic amino acids and amino acid mimetics that do not occur in nature.

[0054] Amino acids may be designated herein by either their commonly known three-letter symbols or the single-letter symbols recommended by the IUPAC-IUB Commission on Biochemical Nomenclature. Nucleotides may also be designated by their common single-letter codes.

[0055] The terms "polypeptide", "peptide" and "protein" are used interchangeably herein to refer to a polymer of amino acid residues, wherein the polymer, in aspects of the present invention, may be conjugated to a moiety that is not composed of amino acids. These terms apply to amino acid polymers in which one or more amino acid residues are an artificial chemical mimetic of the corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers and non-naturally occurring amino acid polymers. The term "fusion protein" refers to a chimeric protein encoding two or more separate protein sequences that are recombinantly expressed as a single fragment.

[0056]The term "conservatively modified options" applies to both amino acid and nucleic acid sequences. With respect to specific nucleic acid sequences, “conservatively modified variants” refer to those nucleic acids that encode identical or substantially identical amino acid sequences. Due to the degeneracy of the genetic code, any given protein will be encoded by a number of nucleic acid sequences. For example, the codons GCA, GCC, GCG, and GCU all code for the amino acid alanine. Thus, at each position at which alanine is specified by a codon, that codon can be changed to any of the corresponding codons described without changing the encoded polypeptide. Such nucleic acid variations are “silent variations,” which are a type of conservatively modified variation. Each nucleic acid sequence herein that encodes a polypeptide also describes all possible silent variations of the nucleic acid. One skilled in the art will appreciate that every codon in a nucleic acid (except AUG, which is typically the only codon for methionine, and TGG, which is typically the only codon for tryptophan) can be modified to form a functionally identical molecule. Accordingly, the presence of each silent variation of the nucleic acid encoding the polypeptide is implied in each sequence described.

[0057] With respect to amino acid sequences, one skilled in the art will understand that single substitutions, deletions, or additions to a nucleic acid, peptide, polypeptide, or protein sequence that alter, add, or delete one amino acid or a small percentage of amino acids in the encoded sequence create thereby a “conservatively modified variant” in which the change results in the replacement of an amino acid with a chemically similar amino acid. Conservative substitution tables containing functionally similar amino acids are well known in the art. Such conservatively modified variants are in addition to and do not exclude polymorphic variants, interspecific homologs and alleles from this description. Each of the following eight groups contains amino acids that are conservative substitutions for each other: (1) alanine (A), glycine (G); (2) aspartic acid (D), glutamic acid (E); (3) asparagine (N), glutamine (Q); (4) arginine (R), lysine (K); (5) isoleucine (I), leucine (L), methionine (M), valine (V); (6) phenylalanine (F), tyrosine (Y), tryptophan (W); (7) serine (S), threonine (T); and (8) cysteine (C), methionine (M) (see, for example, Creighton, Proteins (1984)).

[0058] "Percentage sequence identity" is determined by comparing two optimally aligned sequences in the comparison window, wherein a portion of the polynucleotide or polypeptide sequence in the comparison window may include additions or deletions (i.e., gaps) compared to the reference sequence (which contains no additions or deletions) for optimal alignment of two sequences. This percentage is calculated by determining the number of positions at which an identical nucleic acid base or amino acid residue occurs in both sequences to obtain the number of matching positions, dividing the number of matching positions by the total number of positions in the comparison window and multiplying the result by 100 to obtain the percent identity sequences.

[0059] The terms “identical” or percentage of “identity” in the context of two or more nucleic acid or polypeptide sequences refer to two or more sequences or subsequences that are the same or have a certain percentage of amino acid residues or nucleotides that are the same (i.e. .identity is about 60%, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or better identity in the specified region when compared and aligned for maximum match in the comparison window or in the assigned region) when measured using the BLAST or BLAST 2.0 sequence comparison algorithms with the default parameters described below, or by manual alignment and visual inspection (see, for example, the NCBI website ncbi.nlm.nih.gov/BLAST/ or similar). Such sequences are hereinafter referred to as "substantially identical". This definition also applies or may apply to the complement of the test sequence. This definition also includes sequences with deletions and/or additions, as well as sequences with substitutions. As described below, preferred algorithms may take into account gaps and the like. Preferably, the identity exists in a region that is at least about 25 amino acids or nucleotides in length, or more preferably in a region that is 50-100 amino acids or nucleotides in length.

[0060] The "position" of an amino acid or nucleotide base is designated by a number that sequentially identifies each amino acid (or nucleotide base) in the reference sequence based on its position relative to the N-terminus (or 5'-end). Due to deletions, insertions, truncations, fusions and the like that must be taken into account when determining the optimal alignment, in general the amino acid residue number in the test sequence, determined by simple counting from the N terminus, will not necessarily be the same as the corresponding position number in the reference sequence. For example, in the case where a variant has a deletion relative to an aligned reference sequence, the variant will not have an amino acid at the deletion site that corresponds to a position in the reference sequence. If there is an insertion in an aligned reference sequence, that insertion will not correspond to the numbered amino acid position in the reference sequence. In the case of truncation or fusion, there may be stretches of amino acids in either the reference or aligned sequence that do not match any amino acid in the corresponding sequence.

[0061] The terms “numbered by reference to” or “corresponding to” when used in the context of numbering a particular amino acid or polynucleotide sequence refer to the numbering of residues of the specified reference sequence when the amino acid or polynucleotide sequence is compared to the reference sequence.

[0062] For the specific proteins described herein (e.g., KRAB, dCas9, Dnmt3A, Dnmt3L), the named protein includes any of the naturally occurring forms of the protein, or variants or homologues that retain the activity of the named protein (e.g., in within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to the native protein). In aspects of the invention, the variants or homologues are at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical in amino acid sequence over the entire length of the sequence, or over a region of sequence (e.g., over a contiguous region of 50, 100, 150, or 200 amino acids in length) compared to form found in nature. In aspects of the present invention, the protein is a protein identified by reference to its sequence in NCBI. In aspects of the present invention, a protein is a protein identified by reference to its NCBI sequence, or a functional fragment of such a sequence, or a homologue thereof.

[0063] The term "box-related domain" " or "KRAB domain", as used herein, refers to a category of transcription repressive domains that are present in approximately 400 human zinc finger protein-based transcription factors. KRAB domains typically comprise from about 45 to about 75 amino acid residues. A description of the KRAB domains, including their function and application, can be found, for example, in Ecco, G., Imbeault, M., Trono, D., KRAB zinc finger proteins, Development 144, 2017; Lambert et al. The human transcription factors, Cell 172, 2018; Gilbert et al., Cell (2013); and Gilbert et al., Cell (2014), all of which are incorporated herein by reference in their entirety. In aspects of the present invention, the KRAB domain is a Kox 1 KRAB domain. In aspects of the present invention, the KRAB domain includes the sequence set forth in SEQ ID NO: 16. In aspects of the present invention, the KRAB domain is the sequence set forth in SEQ ID NO: 16. In aspects of the present invention, the KRAB domain includes an amino acid sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 16. In aspects of the present invention, the KRAB domain includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 16. In aspects of the present invention, the KRAB domain includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 16. In aspects of the present invention, the KRAB domain includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 16. In aspects of the present invention, the KRAB domain includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 16. % identical to the sequence of SEQ ID NO: 16. In aspects of the present invention, the KRAB domain includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 16.

[0064] The term "DNA methyltransferase" as used herein refers to an enzyme that catalyzes the transfer of a methyl group to DNA. Non-limiting examples of DNA methyltransferases include Dnmt1, Dnmt3A, Dnmt3B and Dnmt3L. In aspects of the present invention, the DNA methyltransferase is a bacterial cytosine methyltransferase and/or a bacterial nocytosine methyltransferase. Depending on the specific DNA methyltransferase, different parts of the DNA are methylated. For example, Dnmt3A typically targets CpG dinucleotides for methylation. Through DNA methylation, DNA methyltransferases can change the activity of a segment of DNA (such as gene expression) without changing the DNA sequence. In aspects of the present invention, DNA methylation results in repression of gene transcription and/or modulation of methylation-sensitive transcription factors, or CTCFs. As described herein, fusion proteins may include one or more (eg, two) DNA methyltransferases. When a DNA methyltransferase is included as part of a fusion protein, the DNA methyltransferase may be referred to as a “DNA methyltransferase domain.” In aspects of the present invention, a DNA methyltransferase domain includes one or more DNA methyltransferases. In aspects of the present invention, the DNA methyltransferase domain includes two DNA methyltransferases. In aspects of the present invention, the DNA methyltransferase domain is Dnmt3A. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence of SEQ ID NO: 26. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 50%, 55%, 60%, 65%, 70 %, by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98%, is 99% or 100% identical to the sequence of SEQ ID NO: 26. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 26. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 26. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 26. In aspects of this of the invention, the DNA methyltransferase domain has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 26. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 26 In aspects of the present invention, the DNA methyltransferase domain is Dnmt3L. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence of SEQ ID NO: 28. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 50%, 55%, 60%, 65%, 70 %, by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98%, is 99% or 100% identical to the sequence of SEQ ID NO: 28. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 28. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 28. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 28. In aspects of this of the invention, the DNA methyltransferase domain has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 28. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 28 In aspects of the present invention, the DNA methyltransferase domain includes Dnmt3A and Dnmt3L. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence of SEQ ID NO: 33. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 50%, 55%, 60%, 65%, 70 %, by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98%, is 99% or 100% identical to the sequence of SEQ ID NO: 33. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 33. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 33. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 33. In aspects of this of the invention, the DNA methyltransferase domain has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 33. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 33 A description of the structure of the Dnmt3A-3L domain and its application can be found, for example, in Siddique et al, Targeted methylation and gene silencing of VEGF-A in human cells by using a designed Dnmt3a-Dnmt3L single-chain fusion protein with increased DNA methylation activity, J. Mol. Biol. 425, 2013, and in Stepper et al, Efficient targeted DNA methylation with chimeric dCas9-Dnmt3a-Dnmt3L methyltransferase, Nucleic Acids Res. 45, 2017, which are incorporated herein by reference in their entirety and for all purposes.

[0065] The "Dnmt3A", "Dnmt3a", "DNA (cytosine-5) methyltransferase 3A" or "DNA methyltransferase 3a" proteins referred to herein include any of the recombinant or naturally occurring forms of the Dnmt3A enzyme, or variants or homologs thereof that retain Dnmt3A enzyme activity (e.g., to the extent of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92% , 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to Dnmt3A). In aspects of the present invention, variants or homologs are at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical in amino acid sequence over the entire length of the sequence, or at a portion of the sequence ( e.g., over a contiguous stretch of 50, 100, 150, or 200 amino acids in length) compared to the naturally occurring Dnmt3A protein. In aspects of the present invention, the Dnmt3A protein is substantially identical to the protein identified by UniProt accession number Q9Y6K1, or a variant or homolog substantially identical to the protein of that accession number. In aspects of the present invention, the Dnmt3A polypeptide is encoded by a nucleic acid sequence identified by the NCBI reference sequence accession number NM_022552, homologs, or functional fragments of the sequence with this accession number. In aspects of the present invention, Dnmt3A includes the sequence set forth in SEQ ID NO: 26. In aspects of the present invention, Dnmt3A comprises the sequence set forth in SEQ ID NO: 26. In aspects of the present invention, Dnmt3A has an amino acid sequence that is at least 50 %, by 55%, by 60%, by 65%, by 70%, by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, is 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 26. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 75% identical to SEQ ID NO: 26. identical to the sequence of SEQ ID NO: 26. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 26. In aspects of the present invention, the DNA methyltransferase domain has an amino acid sequence that is at least is at least 85% identical to the sequence of SEQ ID NO: 26. In aspects of the present invention, Dnmt3A has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 26. In aspects of the present invention, Dnmt3A has an amino acid sequence that is at least at least 95% identical to the sequence with SEQ ID NO: 26.

[0066] The “Dnmt3L” protein, “DNA (cytosine-5) methyltransferase 3L” or “DNA methyltransferase 3L” as defined herein includes any of the recombinant or naturally occurring forms of the Dnmt3L enzyme, or variants or homologs, that retain Dnmt3L enzyme activity (e.g., within the range of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94 %, 95%, 96%, 97%, 98%, 99% or 100% activity compared to Dnmt3L). In aspects of the invention, the variants or homologues are at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical in amino acid sequence along the entire length of the sequence, or at a portion of the sequence (e.g. on a contiguous stretch of 50, 100, 150, or 200 amino acids in length) compared to the naturally occurring Dnmt3L protein. In aspects of the present invention, the Dnmt3L protein is substantially identical to the protein identified by UniProt accession number Q9CWR8, or a variant or homolog substantially identical to the protein of that accession number. In aspects of the present invention, the Dnmt3L protein is identical to the protein identified by UniProt accession number Q9CWR8. In aspects of the present invention, the Dnmt3L protein sequence is at least 75% identical to the amino acid sequence of the protein identified by UniProt accession number Q9CWR8. In aspects of the present invention, the Dnmt3L protein sequence is at least 80% identical to the amino acid sequence of the protein identified by UniProt accession number Q9CWR8. In aspects of the present invention, the Dnmt3L protein sequence is at least 85% identical to the amino acid sequence of the protein identified by UniProt accession number Q9CWR8. In aspects of the present invention, the Dnmt3L protein sequence is at least 95% identical to the amino acid sequence of the protein identified by UniProt accession number Q9CWR8.

[0067] In aspects of the present invention, the Dnmt3L protein is substantially identical to the protein identified by UniProt accession number Q9UJW, or a variant or homologue substantially identical to the protein of that accession number. In aspects of the present invention, the Dnmt3L protein is identical to the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L protein sequence is at least 50% identical to the amino acid sequence of the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L protein sequence is at least 55% identical to the amino acid sequence of the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L protein sequence is at least 60% identical to the amino acid sequence of the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L protein sequence is at least 65% identical to the amino acid sequence of the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L protein sequence is at least 70% identical to the amino acid sequence of the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L protein sequence is at least 75% identical to the amino acid sequence of the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L protein sequence is at least 80% identical to the amino acid sequence of the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L protein sequence is at least 85% identical to the amino acid sequence of the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L protein sequence is at least 90% identical to the amino acid sequence of the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L protein sequence is at least 95% identical to the amino acid sequence of the protein identified by UniProt accession number Q9UJW. In aspects of the present invention, the Dnmt3L polypeptide is encoded by a nucleic acid sequence identified from the NCBI reference sequence accession number NM_001081695, or from homologs or functional fragments of the sequence with this accession number. In aspects of the present invention, Dnmt3L includes the sequence set forth in SEQ ID NO: 28. In aspects of the present invention, Dnmt3L comprises the sequence set forth in SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 50 %, by 55%, by 60%, by 65%, by 70%, by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, is 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 50% identical to the sequence with SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 55% identical to the sequence with SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 60% identical to the sequence with SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 65% identical to the sequence with SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 97% identical to the sequence with SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 75% identical to the sequence with SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 80% identical to the sequence with SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 85% identical to the sequence with SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 90% identical to the sequence with SEQ ID NO: 28. In aspects of the present invention, Dnmt3L has an amino acid sequence that is at least 95% identical to the sequence with SEQ ID NO: 28.

[0068] The term “RNA-guided DNA endonuclease” and similar terms refer in the ordinary and accepted sense to an enzyme that cleaves a phosphodiester bond in a polynucleotide chain of DNA, wherein recognition of the phosphodiester bond is facilitated by a separate RNA sequence (e.g., a single guide RNA).

[0069] The term “class II CRISPR endonuclease” refers to endonucleases that have the same endonuclease activity as Cas9 and participate in the class II CRISPR system. An example of a class II CRISPR system is the type II CRISPR locus from Streptococcus pyogenes SF370, which contains a cluster of four genes - Cas9, Cas1, Cas2 and Csn1, as well as two non-coding RNA elements - tracrRNA and a characteristic array of repeated sequences (direct repeats) separated by short segments of non-repeating sequences (spacers, about 30 bp each). The Cpf1 enzyme belongs to the putative type V CRISPR-Cas system. Both type II and type V systems are included in class II of the CRISPR-Cas system.

[0070] A "nuclear localization sequence" or "nuclear localization signal" or "NLS" is a peptide that targets proteins to the nucleus. In aspects of the present invention, SNL includes five basic positively charged amino acids. SNL can be located anywhere in the peptide chain. In aspects of the present invention, the SNL is an SV40-derived SNL. In aspects of the present invention, the SNL includes the sequence set forth in SEQ ID NO: 25. In aspects of the present invention, the SNL is the sequence set forth in SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 50 %, by 55%, by 60%, by 65%, by 70%, by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, is 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 75% identical to the sequence with SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 80% identical to the sequence with SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 85% identical with the sequence with SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 90% identical to the sequence with SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 95% identical with the sequence with SEQ ID NO: 25. In aspects of the present invention, the SNL has the amino acid sequence of SEQ ID NO: 25.

[0071] The term “cell” as used herein refers to a cell that performs a metabolic or other function sufficient to maintain or replicate its genomic DNA. A cell can be identified by methods that are well known in the art, including, for example, the presence of an intact membrane, staining with a particular dye, the ability to produce progeny, or, in the case of a gamete, the ability to combine with a second gamete to produce viable progeny. Cells may include prokaryotic and eukaryotic cells. Prokaryotic cells include, but are not limited to, bacteria. Eukaryotic cells include, but are not limited to, yeast cells and cells derived from plants and animals, such as mammalian, insect (eg, Spodoptera ) and human cells. Cells may be useful when they are naturally non-adherent or have been treated so as not to adhere to surfaces, for example by trypsinization.

[0072] The term “vector” as used herein refers to a nucleic acid molecule capable of carrying another nucleic acid to which it has been linked. One type of vector is a "plasmid", which refers to a linear or circular double-stranded loop of DNA into which additional DNA segments can be ligated. Another type of vector is a viral vector, in which additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in the host cell into which they are introduced (eg, bacterial vectors having a bacterial origin of replication and mammalian episomal vectors). Other vectors (eg, non-episomal mammalian vectors) are integrated into the host cell genome after introduction into the host cell and thus replicate along with the host genome. Moreover, certain vectors are capable of controlling the expression of genes to which they are functionally associated. Such vectors are referred to herein as “expression vectors.” In general, expression vectors used in recombinant DNA technologies are often in the form of plasmids. In this document, the terms "plasmid" and "vector" may be used interchangeably, since plasmid is the most commonly used form of vector. However, the present invention is intended to include other forms of expression vectors, such as viral vectors (eg, replication-defective retroviruses, adenoviruses and adeno-associated viruses) that perform equivalent functions. In addition, some viral vectors are capable of targeting a specific cell type either specifically or nonspecifically. Replication-incompetent viral vectors, or replication-defective viral vectors, refer to viral vectors that are able to infect their target cells and deliver their viral load, but then fail to continue the typical lytic pathway, which leads to cell lysis and death.

[0073] The terms "transfection", "transduction", "transfect" or "transduce" can be used interchangeably, and are defined as the process of introducing a nucleic acid molecule and/or protein into a cell. Nucleic acids can be introduced into a cell using non-viral or virus-based methods. The nucleic acid molecule may be a sequence encoding entire proteins or functional parts thereof. Typically, a nucleic acid vector contains elements necessary for protein expression (eg, promoter, transcription start site, etc.). Non-viral transfection methods include any suitable method that does not use viral DNA or viral particles as a delivery system to introduce a nucleic acid molecule into a cell. Typical non-viral transfection methods include encapsulation of nucleic acid nanoparticles that encode the fusion protein (eg, lipid nanoparticles, gold nanoparticles, and the like), calcium phosphate transfection, liposome transfection, nucleofection, sonoporation, heat shock transfection, magnetization transfection, and electroporation . With regard to virus-based methods, any suitable viral vector can be used in the methods described herein. Examples of viral vectors include, but are not limited to, retroviral, adenoviral, lentiviral, and adeno-associated viral vectors. In aspects of the present invention, nucleic acid molecules are introduced into a cell using a retroviral vector in accordance with standard procedures well known in the art. The terms "transfection" or "transduction" also refer to the introduction of proteins into a cell from the external environment. Typically, protein transduction or transfection relies on the attachment of a peptide or protein capable of crossing the cell membrane to the protein of interest. See, for example, Ford et al. (2001) Gene Therapy 8: 1-4, and Prochiantz (2007) Nat. Methods 4: 119-20.

[0074] The term "peptide linker" as used herein is a linker including a peptide moiety. In embodiments of the present invention, the peptide linker is a divalent peptide, such as an amino acid sequence, attached at the N-terminus and C-terminus to the remainder of the compound (eg, a fusion protein provided herein). The peptide linker may be a peptide fragment (a divalent peptide fragment) that can be cleaved (eg, a P2A cleavable polypeptide). A peptide linker as used herein may also be interchangeably referred to as an amino acid linker. In aspects of the present invention, the peptide linker includes from 1 to about 80 amino acid residues. In aspects of the present invention, the peptide linker includes from 1 to about 70 amino acid residues. In aspects of the present invention, the peptide linker includes from 1 to about 60 amino acid residues. In aspects of the present invention, the peptide linker includes from 1 to about 50 amino acid residues. In aspects of the present invention, the peptide linker includes from 1 to about 40 amino acid residues. In aspects of the present invention, the peptide linker includes from 1 to about 30 amino acid residues. In aspects of the present invention, the peptide linker includes from 1 to about 25 amino acid residues. In aspects of the present invention, the peptide linker includes from 1 to about 20 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 20 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 19 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 18 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 17 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 16 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 15 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 14 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 13 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 12 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 11 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 10 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 9 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 8 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 7 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 6 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 5 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 4 amino acid residues. In aspects of the present invention, the peptide linker includes from about 2 to about 3 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 19 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 18 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 17 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 16 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 15 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 14 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 13 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 12 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 11 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 10 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 9 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 8 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 7 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 6 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 5 amino acid residues. In aspects of the present invention, the peptide linker includes from about 3 to about 4 amino acid residues. In aspects of the present invention, the peptide linker includes from about 10 to about 20 amino acid residues. In aspects of the present invention, the peptide linker includes from about 15 to about 20 amino acid residues. In aspects of the present invention, the peptide linker includes about 2 amino acid residues. In aspects of the present invention, the peptide linker includes about 3 amino acid residues. In aspects of the present invention, the peptide linker includes about 4 amino acid residues. In aspects of the present invention, the peptide linker includes about 5 amino acid residues. In aspects of the present invention, the peptide linker includes about 6 amino acid residues. In aspects of the present invention, the peptide linker includes about 7 amino acid residues. In aspects of the present invention, the peptide linker includes about 8 amino acid residues. In aspects of the present invention, the peptide linker includes about 9 amino acid residues. In aspects of the present invention, the peptide linker includes about 10 amino acid residues. In aspects of the present invention, the peptide linker includes about 11 amino acid residues. In aspects of the present invention, the peptide linker includes about 12 amino acid residues. In aspects of the present invention, the peptide linker includes about 13 amino acid residues. In aspects of the present invention, the peptide linker includes about 14 amino acid residues. In aspects of the present invention, the peptide linker includes about 15 amino acid residues. In aspects of the present invention, the peptide linker includes about 16 amino acid residues. In aspects of the present invention, the peptide linker includes about 17 amino acid residues. In aspects of the present invention, the peptide linker includes about 18 amino acid residues. In aspects of the present invention, the peptide linker includes about 19 amino acid residues. In aspects of the present invention, the peptide linker includes about 20 amino acid residues. In aspects of the present invention, the peptide linker includes about 21 amino acid residues. In aspects of the present invention, the peptide linker includes about 22 amino acid residues. In aspects of the present invention, the peptide linker includes about 23 amino acid residues. In aspects of the present invention, the peptide linker includes about 24 amino acid residues. In aspects of the present invention, the peptide linker includes about 25 amino acid residues.

[0075] In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 17. In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 17. In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 18. In aspects of the present invention, the peptide linker is the sequence set forth in SEQ ID NO: 18. In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 19. In aspects of the present invention the peptide linker is the sequence set forth in SEQ ID NO: 19. In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 20. In aspects of the present invention, the peptide linker is the sequence set forth in SEQ ID NO: 20 In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 21. In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 21. In aspects of the present invention, the peptide linker includes the sequence set forth in in SEQ ID NO: 22. In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 22. In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 27. In aspects of the present invention, the peptide linker is the sequence set forth in SEQ ID NO: 27. In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 24. In aspects of the present invention, the peptide linker is the sequence set forth in SEQ ID NO: 24. B In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 29. In aspects of the present invention, the peptide linker is the sequence set forth in SEQ ID NO: 29. In aspects of the present invention, the peptide linker is an XTEN polypeptide. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 17, 18, 19, 20, 21, 22, 24, 27, or 29. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 17, 18, 19, 20, 21, 22, 24, 27 or 29.

[0076] In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 17. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 18. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 19. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to the sequence with SEQ ID NO: 20. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 21. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to sequence of SEQ ID NO: 22. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 24. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 24. % identical to the sequence of SEQ ID NO: 27. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 29. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least is 95% identical to the sequence of SEQ ID NO: 17, 18, 19, 20, 21, 22, 24, 27 or 29. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 17. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 18. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 95% identical to the sequence with SEQ ID NO: 19. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 20. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 95% identical to sequence of SEQ ID NO: 21. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 22. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 22. % identical to the sequence of SEQ ID NO: 24. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 27. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 95% identical to the sequence with SEQ ID NO: 29.

[0077] The terms "XTEN", "XTEN linker" or "XTEN polypeptide" as used herein refer to a recombinant polypeptide (eg, an unstructured recombinant peptide) that does not have hydrophobic amino acid residues. The development and use of XTEN can be found, for example, in Schellenberger et al., Nature Biotechnology 27, 1186-1190 (2009), which is incorporated herein by reference in its entirety and for all purposes. In aspects of the present invention, the XTEN linker includes the sequence set forth in SEQ ID NO: 31. In aspects of the present invention, the XTEN linker includes the sequence set forth in SEQ ID NO: 31. In aspects of the present invention, the XTEN linker includes the sequence set forth in SEQ ID NO: 32. In aspects of the present invention, the XTEN linker is the sequence set forth in SEQ ID NO: 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% , is 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 31. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 31. In aspects of this invention of the invention, the XTEN linker has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 31. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97% , is 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 32. In aspects of this invention The XTEN linker of the invention has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 32.

[0078] A “detectable agent” or “detectable moiety” is a composition that can be detected by appropriate means, such as spectroscopic, photochemical, biochemical, immunochemical, chemical means, magnetic resonance imaging or other physical means. For example, suitable detection reagents include ¹⁸ F, ³² P, ³³ P, ⁴⁵ Ti, ⁴⁷ Sc, ⁵² Fe, ⁵⁹ Fe, ⁶² Cu, ⁶⁴ Cu, ⁶⁷ Cu, ⁶⁷ Ga, ⁶⁸ Ga, ⁷⁷ As, ⁸⁶ Y , ⁹⁰ Y. ⁸⁹ Sr, ⁸⁹ Zr, ⁹⁴ Tc, ⁹⁴ Tc, ^99m Tc, ⁹⁹ Mo, ¹⁰⁵ Pd, ¹⁰⁵ Rh, ¹¹¹ Ag, ¹¹¹ In, ¹²³ I, ¹²⁴ I, ¹²⁵ I, ¹³¹ I, ¹⁴² Pr, ¹⁴³ Pr, ¹⁴⁹ Pm, ¹⁵³ Sm, ^154-1581 Gd, ¹⁶¹ Tb, ¹⁶⁶ Dy, ¹⁶⁶ Ho, ^{169 Er, 175 Lu} , ¹⁷⁷ Lu, ¹⁸⁶ Re, ¹⁸⁸ Re, ¹⁸⁹ Re, ¹⁹⁴ Ir, ¹⁹⁸ Au, ¹⁹⁹ Au, ²¹¹ At, ²¹¹ Pb, ²¹² Bi, ²¹² Pb, ²¹³ Bi, ²²³ Ra, ²²⁵ Ac, Cr, V, Mn, Fe, Co, Ni, Cu, La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, Lu, ³² P, fluorophore (e.g., fluorescent dyes), electron-dense reagents, enzymes (e.g., widely used in ELISA), biotin, digoxigenin, paramagnetic molecules, paramagnetic nanoparticles , ultra-small superparamagnetic iron oxide nanoparticles (USPIO), USPIO nanoparticle aggregates, superparamagnetic iron oxide nanoparticles (SPIO), SPIO nanoparticle aggregates, single-crystalline iron oxide nanoparticles, monocrystalline iron oxide, nanoparticle-based contrast agents, liposomes or other delivery vehicles containing gadolinium chelate molecules (“Gd chelate”) molecules, gadolinium, radioisotopes, radionuclides (e.g., carbon-11, nitrogen-13, oxygen-15, fluorine-18, rubidium-82), fluorodeoxyglucose (e.g., fluorine-18), any gamma-ray emitting radionuclides, positron-emitting radionuclides, radiolabeled glucose, radiolabeled water, radiolabeled ammonia, biocolloids, microbubbles (for example, including microbubble shells containing albumin, galactose, lipids and/or) polymers; inclusive of a gas core of microbubbles containing air, heavy gas(es), perfluorocarbon, nitrogen, octafluoropropane, perflexane lipid microspheres, perflutren, etc.), iodinated contrast agents (e.g. iohexol, iodixanol, ioversol, iopamidol, ioxilan, iopromide, diatrizoate, metrizoate, ioxaglate), barium sulfate, thorium dioxide, gold, gold nanoparticles, gold nanoparticle aggregates, fluorophores, two-photon fluorophores or haptens and proteins, or other objects that can be detected, for example, by incorporating a radiolabel into the peptide or antibody that specifically reacts with the target peptide.

[0079]The detectable moiety is a monovalent detectable agent or a detectable agent capable of forming a bond with another composition. In aspects of the present invention, the detectable agent is an HA tag. In aspects of the present invention, the HA tag includes the sequence set forth in SEQ ID NO: 24. In aspects of the present invention, the HA tag is the sequence set forth in SEQ ID NO: 24. In aspects of the present invention, the HA tag has an amino acid sequence that is at least is at least 80% identical to the sequence of SEQ ID NO: 24. In aspects of the present invention, the HA tag has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 24. In aspects of the present invention, the HA tag has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 24. In aspects of the present invention, the HA tag has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 24. In aspects of the present invention, the detectable agent is blue fluorescent protein (FBP, English "BFP"). In aspects of the present invention, SFB includes the sequence set forth in SEQ ID NO: 30. In aspects of the present invention, SFB is the sequence set forth in SEQ ID NO: 30. In aspects of the present invention, SFB has an amino acid sequence that is at least 80 % identical to the sequence of SEQ ID NO: 30. In aspects of the present invention, SFB has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 30. In aspects of the present invention, SFB has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 30. In aspects of the present invention, SFB has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 30.

[0080] Radioactive substances (e.g., radioisotopes) that can be used as imaging and/or labeling agents in accordance with aspects of the present invention include, but are not limited to, ¹⁸ F, ³² P, ³³ P, ⁴⁵ Ti, ⁴⁷ Sc ^{, 52} Fe, ⁵⁹ Fe, ⁶² Cu, ⁶⁴ Cu, 67 Cu, ⁶⁷ Ga, ⁶⁸ Ga, ⁷⁷ As, ⁸⁶ Y, ⁹⁰ Y, ⁸⁹ Sr, ⁸⁹ Zr, ⁹⁴ Tc, ⁹⁴ Tc, ^99m Tc, ⁹⁹ Mo, ¹⁰⁵ Pd, ¹⁰⁵ Rh, ¹¹¹ Ag, ¹¹¹ In, ¹²³ I, ¹²⁴ I, ¹²⁵ I, ¹³¹ I, ¹⁴² Pr, ¹⁴³ Pr, ¹⁴⁹ Pm, ¹⁵³ Sm, ^154-1581 Gd, ¹⁶¹ Tb, ¹⁶⁶ Dy, ¹⁶⁶ Ho, ¹⁶⁹ Er, ¹⁷⁵ Lu, ¹⁷⁷ Lu, ^{186 Re, 188 Re} , ¹⁸⁹ Re, ¹⁹⁴ Ir, ¹⁹⁸ Au, ¹⁹⁹ Au, ²¹¹ At, ²¹¹ Pb, ²¹² Bi, ²¹² Pb, ²¹³ Bi, ²²³ Ra and ²²⁵ Ac . Paramagnetic ions that can be used as additional imaging agents in accordance with aspects of this invention include, but are not limited to, transition metal and lanthanide metal ions (e.g., metals with atomic numbers 21-29, 42, 43, 44 or 57-71). These metal ions include Cr, V, Mn, Fe, Co, Ni, Cu, La, Ce, Pr, Nd, Pm, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb and Lu.

[0081]Term "contacting" is used in its simple, ordinary meaning and refers to the process of allowing at least two different molecules to come close enough to each other to react, interact, or physically touch. However, it should be understood that the final reaction product may be obtained directly from the reaction between the added reactants, or from an intermediate product of one or more added reactants that may be obtained in the reaction mixture.

[0082] The term "contacting" may include allowing two molecules to react, interact, or physically touch, where the two molecules may be, for example, a fusion protein as provided herein and a nucleic acid sequence (e.g., a target DNA sequence ).

[0083] As defined herein, the term “inhibiting,” “inhibits,” “inhibiting,” “repressing,” “represses,” “silencing,” “silencing,” and the like when used in relation to a composition provided herein document (e.g., fusion protein, complex, nucleic acid, vector) refer to the negative effect (e.g., decrease) of the activity (e.g., transcription) of a nucleic acid sequence (e.g., decrease in gene transcription) relative to the activity of the nucleic acid sequence (e.g., gene transcription ) in the absence of a composition (eg, fusion protein, complex, nucleic acid, vector). In aspects of the present invention, inhibition refers to the attenuation of a disease or symptoms of a disease (eg, cancer). Thus, inhibition involves, at least in part, partially or completely blocking the activity (eg, transcription) or reducing, preventing or delaying the activity (eg, transcription) of a nucleic acid sequence. Inhibited activity (eg, transcription) may be 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, or less than the activity in the control. In aspects of the present invention, the inhibition is 1.5-fold, 2-fold, 3-fold, 4-fold, 5-fold, 10-fold or more compared to the control.

[0084] A “control” sample or value refers to a sample that serves as a reference, usually a known reference, for comparison with the test sample. For example, a test sample may be taken from a test condition, eg in the presence of a test compound, and compared with samples from known conditions, eg in the absence of a test compound (negative control) or in the presence of a known compound (positive control). A control may also be an average obtained from a number of studies or results. One skilled in the art will appreciate that controls can be designed to assess any number of parameters. For example, controls can be developed to compare therapeutic effect based on pharmacological data (eg, half-life) or therapeutic measures (eg, comparison of side effects). One skilled in the art will understand which controls are important in a particular situation and will be able to analyze data based on comparisons with control values. Controls are also important to determine the significance of the data. For example, if values for a particular parameter are very different in controls, then the variation in the test samples will not be considered significant.

[0085] Fusion proteins

[0086] Presented herein are, among other things, fusion proteins that can disable genes permanently (eg, irreversibly) and reversibly in mammalian cells using CRISPR-based epigenome editing. In embodiments of the present invention, the fusion protein includes a single polypeptide fused from four proteins (eg, catalytically inactive Cas9 (eg, dCas9), KRAB domain, Dnmt3A, and Dnmt3L) that can be transiently expressed in cells. The fusion protein can be directed to a specific site in the genome of a mammal using a polynucleotide that is complementary to the target nucleic acid sequence (eg, a DNA sequence) and which further includes a sequence (ie, a binding sequence) capable of binding to the fusion protein. Once correctly positioned, and without intending to be bound by theory, the fusion protein adds DNA methylation markers and/or repressive chromatin markers to the target nucleic acid, resulting in gene silencing that is inherited through subsequent cell divisions. Thus, the fusion protein can perform epigenome editing that bypasses the creation of DNA double-strand breaks in the host genome, making it a safe and reversible way to manipulate the genome of a living organism.

[0087] In embodiments of the present invention, the fusion protein comprises a nuclease-deficient RNA-guided DNA endonuclease enzyme; KRAB domain and DNA methyltransferase domain. In aspects of the present invention, the fusion protein comprises, from N-terminus to C-terminus, a DNA methyltransferase domain, a nuclease-deficient RNA-guided DNA endonuclease enzyme, and a KRAB domain. In aspects of the present invention, the fusion protein comprises, from N-terminus to C-terminus, a nuclease-deficient RNA-guided DNA endonuclease enzyme, and a DNA methyltransferase domain. In embodiments of the present invention, the fusion protein further comprises one or more peptide linkers. In aspects of the present invention, the fusion protein further comprises one or more detectable tags. In aspects of the present invention, the fusion protein further comprises one or more nuclear localization sequences. In aspects of the present invention, the fusion protein further comprises one or more peptide linkers, one or more detectable tags, one or more nuclear localization sequences, or a combination of two or more of the above. When the fusion protein contains one or more peptide linkers, each of the peptide liners may be the same or different. When the fusion protein contains one or more detectable tags, each of the detectable tags may be the same or different. In aspects of the present invention, the fusion protein contains from 1 to 10 detectable tags. In aspects of the present invention, the fusion protein contains from 1 to 9 detectable tags. In aspects of the present invention, the fusion protein contains from 1 to 8 detectable tags. In aspects of the present invention, the fusion protein contains from 1 to 7 detectable tags. In aspects of the present invention, the fusion protein contains from 1 to 6 detectable tags. In aspects of the present invention, the fusion protein contains from 1 to 5 detectable tags. In aspects of the present invention, the fusion protein contains from 1 to 4 detectable tags. In aspects of the present invention, the fusion protein contains from 1 to 3 detectable tags. In aspects of the present invention, the fusion protein contains 1 to 2 detectable tags. In aspects of the present invention, the fusion protein contains 1 detectable tag. In aspects of the present invention, the fusion protein contains 2 detectable tags. In aspects of the present invention, the fusion protein contains 3 detectable tags. In aspects of the present invention, the fusion protein contains 4 detectable tags. In aspects of the present invention, the fusion protein contains 5 detectable tags.

[0088] In embodiments of the present invention, the fusion protein contains the structure A - B - C, or B - A - C, or C - A - B, or C - B - A, or B - C - A, or A - C - B; where A is an enzyme nuclease-deficient RNA-directed DNA endonuclease; B is a KRAB domain; C is a DNA methyltransferase domain; and wherein the component on the left is the N-terminus and the component on the right is the C-terminus. In aspects of the present invention, the fusion protein further comprises one or more peptide linkers and one or more detectable tags. In aspects of the present invention, A - B, B - A, B - C, C - B, A - C, and C - A are each independently linked to each other by a covalent bond, a peptide linker, a detectable tag, a nuclear localization sequence, or a combination of two or more of them. The peptide linker can be any known in the art (eg, P2A cleavable peptide, XTEN linker, and the like). In aspects of the present invention, the fusion protein contains other components, such as detectable tags (eg, HA tag, blue fluorescent protein, etc.).

[0089] In embodiments of the present invention, the fusion protein comprises the structure A - L ₁ - B - L ₂ - C, wherein A is an enzyme nuclease-deficient RNA-guided DNA endonuclease; B is a KRAB domain, C is a DNA methyltransferase domain, L ₁ is a covalent bond or peptide linker, and L ₂ is a covalent bond or peptide linker; and where A is at the N-terminus and C is at the C-terminus. In aspects of the present invention, A is covalently linked to B through a peptide linker. In aspects of the present invention, A is covalently linked to B through a covalent bond. In aspects of the present invention, B is covalently linked to C through a peptide linker. In aspects of the present invention, B is covalently linked to C through a covalent bond. The peptide linker can be any known in the art (eg, P2A cleavable peptide, XTEN linker, and the like). In aspects of the present invention, the fusion protein contains other components, such as detectable tags, nuclear localization sequences, and the like. In aspects of the present invention, L ₁ is a covalent bond, a peptide linker, a detectable tag, a nuclear localization sequence, or a combination thereof. In aspects of the present invention, L ₂ is a covalent bond, a peptide linker, a detectable tag, a nuclear localization sequence, or a combination thereof.

[0090] In embodiments of the present invention, the fusion protein comprises the structure B - L ₁ - A - L ₂ - C, wherein A is an enzyme nuclease-deficient RNA-guided DNA endonuclease; B is a KRAB domain, C is a DNA methyltransferase domain, L ₁ is a covalent bond or peptide linker, and L ₂ is a covalent bond or peptide linker; and where B is at the N-terminus and C is at the C-terminus. In aspects of the present invention, L ₁ is a peptide linker. In aspects of the present invention, L ₁ represents a covalent bond. In aspects of the present invention, L ₂ is a peptide linker. In aspects of this invention, L ₂ represents a covalent bond. The peptide linker can be any known in the art or described herein (eg, P2A cleavable peptide, XTEN linker, etc.). In aspects of the present invention, the fusion protein contains other components, such as detectable tags. In aspects of the present invention, L ₁ is a covalent bond, a peptide linker, a detectable label, or a combination thereof. In aspects of the present invention, L ₂ is a covalent bond, a peptide linker, a detectable label, or a combination thereof. In aspects of the present invention, the fusion protein further comprises a nuclear localization sequence. Exemplary fusion proteins containing the B - L ₁ - A - L ₂ - C structure include p76, p90, p91, p92, p93, p94, p95, p96, p97, p98, p99, p100, p101 and p102 (Fig. . 5)

[0091] In embodiments of the present invention, the fusion protein comprises the structure B - L ₃ - A - L ₄ - C - L ₅ - D; where A is an enzyme nuclease-deficient RNA-guided DNA endonuclease; B is a KRAB domain, C is a DNA methyltransferase domain, D is absent or D is one or more detectable tags, L ₃ is a covalent bond, a peptide linker, a detectable tag or a combination of two or more of these, L ₄ is a covalent a bond, a peptide linker, a detectable label, or a combination of two or more of them, L ₅ is absent or L ₅ is a covalent bond or peptide linker; and where B is at the N-terminus and D is at the C-terminus. In aspects of the present invention, L ₃ is a peptide linker. In aspects of this invention, L ₃ represents a covalent bond. In aspects of the present invention, L ₃ contains a peptide linker and a detectable label. In aspects of the present invention, L ₃ contains a detectable label. In aspects of the present invention, L ₄ is a peptide linker. In aspects of the present invention, L ₄ contains a peptide linker and a detectable label. In aspects of this invention, L ₄ represents a covalent bond. In aspects of the present invention, L ₄ contains a detectable label. In aspects of the present invention, L ₅ is a peptide linker. In aspects of this invention, L ₅ represents a covalent bond. In aspects of the present invention, D contains one or more detectable labels. In aspects of the present invention, D comprises a single detectable label. In aspects of the present invention, D contains two detectable labels. In aspects of the present invention, D contains three detectable labels. In aspects of the present invention, D contains a plurality of detectable labels. D may be any detectable label known in the art and/or described herein (eg, HA tag, blue fluorescent protein, etc.). In aspects of the present invention, L ₅ and D may be absent. When L ₃ , L ₄ , L ₅ and D contain two or more detectable marks, each of the detected marks is the same or different. The peptide linker can be any known in the art and/or described herein (eg, P2A cleavable peptide, XTEN linker, etc.). In aspects of the present invention, the fusion protein further comprises a nuclear localization sequence. Exemplary fusion proteins containing the structure B - L ₃ - A - L ₄ - C - L ₅ - D include p76, p90, p91, p92, p93, p94, p95, p96, p97, p98, p99, p100, p101 and p102, as shown in FIG. 5.

[0092] In embodiments of the present invention, the fusion protein comprises the structure C - L ₃ - A - L ₄ - B - L ₅ - D, wherein A is an enzyme nuclease-deficient RNA-guided DNA endonuclease; B is a KRAB domain, C is a DNA methyltransferase domain, D is absent or D contains one or more detectable tags, L ₃ is a covalent bond, a peptide linker, a detectable tag or a combination of two or more of these, L ₄ is a covalent bond , a peptide linker, a detectable label, or a combination of two or more of them, L ₅ is absent or L ₅ is a covalent bond or a peptide linker; and where C is at the N-terminus and D is at the C-terminus. In aspects of the present invention, L ₃ is a peptide linker. In aspects of this invention, L ₃ represents a covalent bond. In aspects of the present invention, L ₃ contains a detectable label. In aspects of the present invention, L ₃ contains a peptide linker and a detectable label. In aspects of the present invention, L ₄ is a peptide linker. In aspects of this invention, L ₄ represents a covalent bond. In aspects of the present invention, L ₄ contains a detectable label. In aspects of the present invention, L ₄ contains a peptide linker and a detectable label. In aspects of the present invention, L ₅ is a peptide linker. In aspects of this invention, L ₅ represents a covalent bond. In aspects of the present invention, D contains one or more detectable labels. In aspects of the present invention, D comprises a single detectable label. In aspects of the present invention, D contains two detectable labels. In aspects of the present invention, D contains three detectable labels. In aspects of the present invention, D contains a plurality of detectable labels. D may be any detectable label known in the art and/or described herein (eg, HA tag, blue fluorescent protein, etc.). In aspects of the present invention, L ₅ and D may be absent. When L ₃ , L ₄ , L ₅ and D contain two or more detectable marks, each of the detected marks is the same or different. The peptide linker can be any known in the art and/or described herein (eg, P2A cleavable peptide, XTEN linker, etc.). In aspects of the present invention, the fusion protein further comprises a nuclear localization sequence. Exemplary fusion proteins containing the structure C - L ₃ - A - L ₄ - B - L ₅ - D include p112, as shown in FIG. 5.

[0093] The term “enzyme nuclease-deficient RNA-guided DNA endonuclease” and the like refers in the ordinary and accepted sense to an RNA-guided DNA endonuclease (e.g., a mutant form of a naturally occurring RNA-guided DNA endonuclease) that targets a specific phosphodiester bond in a DNA polynucleotide where recognition of the phosphodiester bond is facilitated by a distinct polynucleotide sequence (e.g., an RNA sequence (e.g., single guide RNA (sgRNA))) but which is not capable of cleaving the target phosphodiester bond to a significant extent (e.g., there is no measurable cleavage of the phosphodiester bond under physiological conditions).The nuclease-deficient RNA-guided DNA endonuclease thus retains DNA-binding capacity (e.g., specific binding to a target sequence) when complexed with a polynucleotide (e.g., egRNA), but does not have significant endonuclease activity (e.g. , any detectable level of endonuclease activity). In aspects of the present invention, the nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9, ddCpf1, a nuclease-deficient variant of Cas9, or a class II nuclease-deficient CRISPR endonuclease.

[0094] In embodiments of the present invention, the nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9. The terms “dCas9” or “dCas9 protein” as used herein refer to a Cas9 protein in which both catalytic sites for endonuclease activity are defective or inactive. In aspects of the present invention, the dCas9 protein has mutations at positions corresponding to D10A and H840A in Cas9 from S. pyogenes . In aspects of the present invention, the dCas9 protein lacks endonuclease activity due to point mutations in both catalytic endonuclease sites (RuvC and HNH) of wild-type Cas9. Point mutations may be D10A and H840A. In aspects of the present invention, dCas9 has substantially no detectable endonuclease (eg, endodeoxyribonuclease) activity. In aspects of the present invention, dCas9 includes the amino acid sequence of SEQ ID NO: 23. In aspects of the present invention, dCas9 has the amino acid sequence of SEQ ID NO: 23. In aspects of the present invention, dCas9 has an amino acid sequence that is at least 50% 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% , is 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 23. In aspects of the present invention, dCas9 has an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 23. In aspects of the present invention, dCas9 has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 23. In aspects of the present invention, dCas9 has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 23. In aspects of the present invention, dCas9 has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 23. In aspects of the present invention, dCas9 has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 23.

[0095] The terms “CRISPR-associated protein 9,” “Cas9,” “Csn1,” or “Cas9 protein,” as used herein, include any of the recombinant or naturally occurring forms of the Cas9 endonuclease, or variants or homologs thereof, that retain the endonuclease activity of the Cas9 enzyme (e.g., within the range of at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% activity compared to Cas9). In some aspects of the invention, variants or homologs are at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical in amino acid sequence over the entire length of the sequence, or at a region of the sequence (e.g., over a contiguous region of 50, 100, 150, or 200 amino acids in length) compared to the naturally occurring Cas9 protein. In aspects of the present invention, the Cas9 protein is substantially identical to the protein identified by UniProt accession number Q99ZW2, or a variant or homologue substantially identical to the protein of that accession number. In aspects of the present invention, the Cas9 protein sequence is at least 75% identical to the amino acid sequence of the protein identified by UniProt accession number Q99ZW2. In aspects of the present invention, the Cas9 protein sequence is at least 80% identical to the amino acid sequence of the protein identified by UniProt accession number Q99ZW2. In aspects of the present invention, the Cas9 protein sequence is at least 85% identical to the amino acid sequence of the protein identified by UniProt accession number Q99ZW2. In aspects of the present invention, the Cas9 protein sequence is at least 90% identical to the amino acid sequence of the protein identified by UniProt accession number Q99ZW2. In aspects of the present invention, the Cas9 protein sequence is at least 95% identical to the amino acid sequence of the protein identified by UniProt accession number Q99ZW2.

[0096] In embodiments of the present invention, the nuclease-deficient RNA-guided DNA endonuclease enzyme is “ddCpf1” or “ddCas12a”. The terms "DNase-dead Cpf1" or "ddCpf1" refer to Cpf1 from Acidaminococcus sp . (AsCpf1), containing a mutation that leads to inactivation of DNase activity in Cpf1. In aspects of the present invention, ddCpf1 includes the E993A mutation in the RuvC domain of AsCpf1. In aspects of the present invention, ddCpf1 has substantially no detectable endonuclease (eg, endodeoxyribonuclease) activity. In aspects of the present invention, ddCpf1 includes the amino acid sequence of SEQ ID NO: 34. In aspects of the present invention, ddCpf1 has the amino acid sequence of SEQ ID NO: 34. In aspects of the present invention, ddCpf1 has an amino acid sequence that is at least 50% 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% , is 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 34. In aspects of the present invention, ddCpf1 has an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 34. In aspects of the present invention, ddCpf1 has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 34. In aspects of the present invention, ddCpf1 has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 34. In aspects of the present invention, ddCpf1 has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 34. In aspects of the present invention, ddCpf1 has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 34.

[0097] In embodiments of the present invention, the nuclease-deficient RNA-guided DNA endonuclease enzyme is dLbCpf1. The term "dLbCpf1" refers to mutated Cpf1 from the bacterium Lachnospiraceae ND2006 (LbCpf1), which lacks DNase activity. In aspects of the present invention, dLbCpf1 includes the D832A mutation. In aspects of the present invention, dLbCpf1 has substantially no detectable endonuclease (eg, endodeoxyribonuclease) activity. In aspects of the present invention, dLbCpf1 includes the amino acid sequence of SEQ ID NO: 35. In aspects of the present invention, dLbCpf1 has the amino acid sequence of SEQ ID NO: 35. In aspects of the present invention, dLbCpf1 has an amino acid sequence that is at least 50% 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% , is 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 35. In aspects of the present invention, dLbCpf1 has an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 35. In aspects of the present invention, dLbCpf1 has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 35. In aspects of the present invention, dLbCpf1 has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 35. In aspects of the present invention, dLbCpf1 has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 35. In aspects of the present invention, dLbCpf1 has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 35.

[0098] In embodiments of the present invention, the nuclease-deficient RNA-guided DNA endonuclease enzyme is dFnCpf1. The term "dFnCpf1" refers to the mutated Cpf1 from Francisella novicida U112 (FnCpf1), which lacks DNase activity. In aspects of the present invention, dFnCpf1 includes the D917A mutation. In aspects of the present invention, dFnCpf1 has substantially no detectable endonuclease (eg, endodeoxyribonuclease) activity. In aspects of the present invention, dFnCpf1 includes the amino acid sequence of SEQ ID NO: 36. In aspects of the present invention, dFnCpf1 has the amino acid sequence of SEQ ID NO: 36. In aspects of the present invention, dFnCpf1 has an amino acid sequence that is at least 50% 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95% , is 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 36. In aspects of the present invention, dFnCpf1 has an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 36. In aspects of the present invention, dFnCpf1 has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 36. In aspects of the present invention, dFnCpf1 has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 36. In aspects of the present invention, dFnCpf1 has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 36. In aspects of the present invention, dFnCpf1 has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 36.

[0099] The term "Cpf1" or "Cpf1 protein" as used herein includes any of the recombinant or naturally occurring forms of the Cpf1 endonuclease (CRISPR from Prevotella and Francisella 1), or variants or homologues thereof that retain endonuclease activity of the Cpf1 enzyme (eg, within the range of at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to Cpf1). In aspects of the present invention, variants or homologs are at least 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical in amino acid sequence over the entire length of the sequence, or at a portion of the sequence ( e.g., over a contiguous stretch of 50, 100, 150, or 200 amino acids in length) compared to the naturally occurring Cpf1 protein. In aspects of the present invention, the Cpf1 protein is substantially identical to the protein identified by UniProt accession number U2UMQ6, or a variant or homolog substantially identical to the protein of that accession number. In aspects of the present invention, the Cpf1 protein is identical to the protein identified by UniProt accession number U2UMQ6. In aspects of the present invention, the Cpf1 protein sequence is at least 75% identical to the amino acid sequence of the protein identified by UniProt accession number U2UMQ6. In aspects of the present invention, the Cpf1 protein sequence is at least 80% identical to the amino acid sequence of the protein identified by UniProt accession number U2UMQ6. In aspects of the present invention, the Cpf1 protein is identical to the protein identified by UniProt accession number U2UMQ6. In aspects of the present invention, the Cpf1 protein sequence is at least 85% identical to the amino acid sequence of the protein identified by UniProt accession number U2UMQ6. In aspects of the present invention, the Cpf1 protein is identical to the protein identified by UniProt accession number U2UMQ6. In aspects of the present invention, the Cpf1 protein sequence is at least 90% identical to the amino acid sequence of the protein identified by UniProt accession number U2UMQ6. In aspects of the present invention, the Cpf1 protein is identical to the protein identified by UniProt accession number U2UMQ6. In aspects of the present invention, the Cpf1 protein sequence is at least 95% identical to the amino acid sequence of the protein identified by UniProt accession number U2UMQ6.

[0100] In embodiments of the present invention, the nuclease-deficient RNA-guided DNA endonuclease enzyme is a nuclease-deficient variant of Cas9. The term “nuclease-deficient Cas9 variant” refers to a Cas9 protein that has one or more mutations that increase its PAM binding specificity compared to wild-type Cas9, and in addition has mutations that make a protein incapable of endonuclease activity or having seriously impaired endonuclease activity. Without being limited by theory, it is believed that the target sequence should be associated with a PPM (protospacer adjacent motif); that is, it is a short sequence that is recognized by the CRISPR complex. The exact sequence and length requirements for PPMs vary depending on the CRISPR enzyme used, but PPMs are typically 2-5 base pair sequences adjacent to the protospacer (i.e., the target sequence). The binding specificity of nuclease-deficient Cas9 variants to PPM can be determined by any method known in the art. A description and application of known Cas9 variants can be found, for example, in Shmakov et al., Diversity and evolution of class 2 CRISPR-Cas systems. Nat. Rev. Microbiol. 15, 2017, and in Cebrian-Serrano et al, CRISPR-Cas orthologues and variants: optimizing the repertoire, specificity and delivery of genome engineering tools. Mamm. Genome 7-8, 2017, which are incorporated herein by reference in their entirety and for all purposes. Exemplary Cas9 variants are listed in Table 4 below.

[0101] Table 4

Cas9 variants MRP domains Literary sources Streptococcus pyogenes (Sp) Cas9 NGG Hsu et al. 2014 Cell Staphylococcus aureus (Sa)Cas9 NNGRRT or NNGRR NNGGGT, NNGAAT, NNGAGT (Zetsche) Ran et al. 2015 Nature SpCas9 VQR mutant (D1135V, R1335Q, T1337R) NGAG > NGAT=NGAA > NGAC NGCG Kleinstiver et al. 2015 Nature SpCas9 VRER mutant (D1135V/ G1218R/ R1335E/ T1337R) NGCG Kleinstiver et al. 2015 Nature SpCas9 D1135E NGG, more accuracy, fewer abbreviations on NAG and NGA sites Kleinstiver et al. 2015 Nature Mutant eSpCas9 1.1 (K848A/ K1003A/ R1060A) NGG Slaymaker et al. Science 2015 SpCas9HF1
(Q695A, Q926A, N497A, R661A) NGG Kleinstiver et al. 2016 Nature AsCpf1 TTTN (5' in egRNA) Zetsche et al. 2015 Cell HypaCas9 (N692A, M694A, Q695A, H698A) Chen et al., Nature, volume 550, pages 407-410 (19 October 2017)

[0102] In embodiments of the present invention, the nuclease-deficient RNA-guided DNA endonuclease enzyme is a class II nuclease-deficient CRISPR endonuclease. The term “class II nuclease-deficient CRISPR endonuclease” as used herein refers to any class II CRISPR endonuclease that has mutations resulting in reduced, impaired, or complete loss of endonuclease activity.

[0103] In embodiments of the present invention, the DNA methyltransferase domain is a Dnmt3A-3L domain. The term “Dnmt3A-3L domain” as used herein refers to a protein comprising both Dnmt3A and Dnmt3L. In aspects of the present invention, Dnmt3A and Dnmt3L are covalently linked. In some aspects of the present invention, Dnmt3A is covalently linked to Dnmt3L via a peptide linker. In aspects of the present invention, the peptide linker includes the sequence set forth in SEQ ID NO: 27. In aspects of the present invention, the peptide linker is the sequence set forth in SEQ ID NO: 27. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least at least by 50%, by 55%, by 60%, by 65%, by 70%, by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 27. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 27. In aspects of the present invention, the peptide linker has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 27. In aspects of the present invention, the Dnmt3A-3L domain includes the sequence, set forth in SEQ ID NO: 33. In aspects of the present invention, the Dnmt3A-3L domain is the sequence set forth in SEQ ID NO: 33. In aspects of the present invention, the Dnmt3A-3L domain has an amino acid sequence that is at least 50% %, by 60%, by 65%, by 70%, by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 33. In aspects of the present invention, the Dnmt3A-3L domain has an amino acid sequence that is at least 75% identical to the sequence with SEQ ID NO: 33. In aspects of the present invention, the Dnmt3A-3L domain has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 33. In aspects of the present invention, the Dnmt3A-3L domain has an amino acid sequence that is at least is 85% identical to the sequence of SEQ ID NO: 33. In aspects of the present invention, the Dnmt3A-3L domain has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 33. In aspects of the present invention, the Dnmt3A-3L domain has an amino acid sequence a sequence that is at least 95% identical to the sequence of SEQ ID NO: 33.

[0104] In embodiments of the present invention, the peptide linker is an XTEN linker. In aspects of the present invention, the XTEN linker includes from about 16 to about 80 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 17 to about 80 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 18 to about 80 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 19 to about 80 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 20 to about 80 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 30 to about 80 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 40 to about 80 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 50 to about 80 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 60 to about 80 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 70 to about 80 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 16 to about 70 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 16 to about 60 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 16 to about 50 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 16 to about 40 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 16 to about 35 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 16 to about 30 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 16 to about 25 amino acid residues. In aspects of the present invention, the XTEN linker includes from about 16 to about 20 amino acid residues. In aspects of the present invention, the XTEN linker includes about 16 amino acid residues. In aspects of the present invention, the XTEN linker includes about 17 amino acid residues. In aspects of the present invention, the XTEN linker includes about 18 amino acid residues. In aspects of the present invention, the XTEN linker includes about 19 amino acid residues. In aspects of the present invention, the XTEN linker includes about 20 amino acid residues.

[0105] In embodiments of the present invention, the XTEN linker includes the sequence set forth in SEQ ID NO: 31. In aspects of the present invention, the XTEN linker includes the sequence set forth in SEQ ID NO: 31. In aspects of the present invention, the XTEN linker includes sequence set forth in SEQ ID NO: 32. In aspects of the present invention, the XTEN linker is the sequence set forth in SEQ ID NO: 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96% , is 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 31. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 31 In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 31. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 31. : 31. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 32 In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 32. : 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 32.

[0106] The fusion protein may include amino acid sequences suitable for targeting the fusion protein to specific regions of the cell (eg, cytoplasm, nucleus). Thus, in aspects of the present invention, the fusion protein further includes a nuclear localization signal (NLS) peptide. In aspects of the present invention, the SNL includes the sequence set forth in SEQ ID NO: 25. In aspects of the present invention, the SNL is the sequence set forth in SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 50 %, by 55%, by 60%, by 65%, by 70%, by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, is 95%, 96%, 97%, 98%, 99%, or 100% identical to the sequence of SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 75% identical to the sequence with SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 80% identical to the sequence with SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 85% identical to the sequence with SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 90% identical to the sequence with SEQ ID NO: 25. In aspects of the present invention, the SNL has an amino acid sequence that is at least 95% identical to the sequence with SEQ ID NO: 25.

[0107] In embodiments of the present invention, the fusion protein includes, in an N-terminal to C-terminal direction, a KRAB domain, a nuclease-deficient RNA-guided DNA endonuclease enzyme, and a DNA methyltransferase domain.

[0108] In embodiments of the present invention, the nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9 and the DNA methyltransferase domain is Dnmt3A-3L domain.

[0109] In embodiments of the present invention, dCas9 is covalently linked to the KRAB domain via a peptide linker, and wherein dCas9 is covalently linked to the Dnmt3A-3L domain via a peptide linker.

[0110] In embodiments of the present invention, the peptide linker is an XTEN linker. In aspects of the present invention, the XTEN linker includes the sequence set forth in SEQ ID NO: 31. In aspects of the present invention, the XTEN linker is the sequence set forth in SEQ ID NO: 31. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least is at least 75% identical to the sequence of SEQ ID NO: 31. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 31. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 31. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 31. In aspects of the present invention, the XTEN linker has an amino acid sequence which is at least 95% identical to the sequence set forth in SEQ ID NO: 31. In aspects of the present invention, the XTEN linker includes the sequence set forth in SEQ ID NO: 32. In aspects of the present invention, the XTEN linker includes the sequence set forth in SEQ ID NO: : 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 90% identical to the sequence with SEQ ID NO: 32. In aspects of the present invention, the XTEN linker has an amino acid sequence that is at least 95% identical to the sequence with SEQ ID NO: 32.

[0111] In embodiments of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 1. In aspects of the present invention, the fusion protein is an amino acid sequence of SEQ ID NO: 1. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: : 2. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 2. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 3. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 3. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 4. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 4. In aspects of the present invention, the fusion protein includes the amino acid the sequence of SEQ ID NO: 5. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 5. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 6. In aspects of the present invention, the fusion protein is is the amino acid sequence of SEQ ID NO: 6. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 7. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 7. In aspects of the present invention, the fusion protein is the protein includes the amino acid sequence of SEQ ID NO: 8. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 8. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 9. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 9. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 10. In aspects of the present invention, the fusion protein is of the amino acid sequence of SEQ ID NO: 10. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 11. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 11. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: : 12. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 12. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 13. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 13. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 14. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 14. In aspects of the present invention, the fusion protein includes the amino acid sequence of SEQ ID NO: 15. In aspects of the present invention, the fusion protein is the amino acid sequence of SEQ ID NO: 15.

[0112] In embodiments of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86 %, by 87%, by 88%, by 89%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98% or is 99% identical to the sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In aspects of the present invention, the fusion protein includes an amino acid a sequence that is at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 1. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identical to SEQ ID NO: 2. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, by 86%, by 87%, by 88%, by 89%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98 % or 99% identical to the sequence of SEQ ID NO: 3. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, 82%, 83%, by 84%, by 85%, by 86%, by 87%, by 88%, by 89%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96 %, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 4. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, by 82%, by 83%, by 84%, by 85%, by 86%, by 87%, by 88%, by 89%, by 90%, by 91%, by 92%, by 93%, by 94 %, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 5. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to SEQ ID NO: 5. by 80%, by 81%, by 82%, by 83%, by 84%, by 85%, by 86%, by 87%, by 88%, by 89%, by 90%, by 91%, by 92 %, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 6. In aspects of the present invention, the fusion protein includes an amino acid sequence, which is at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90 %, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 7. In aspects of the present invention the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88 %, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence with SEQ ID NO: 8. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86 %, by 87%, by 88%, by 89%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98% or is 99% identical to the sequence of SEQ ID NO: 9. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, 82%, 83%, 84 %, by 85%, by 86%, by 87%, by 88%, by 89%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, is 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 10. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, 82 %, by 83%, by 84%, by 85%, by 86%, by 87%, by 88%, by 89%, by 90%, by 91%, by 92%, by 93%, by 94%, is 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 11. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80 %, by 81%, by 82%, by 83%, by 84%, by 85%, by 86%, by 87%, by 88%, by 89%, by 90%, by 91%, by 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 12. In aspects of the present invention, the fusion protein includes an amino acid sequence that at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 13. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence with SEQ ID NO: 14. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, by 87%, by 88%, by 89%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98% or by 99 % identical to the sequence with SEQ ID NO: 15.

[0113] In embodiments of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 1. In aspects of the present invention, the fusion protein includes an amino acid sequence, which is at least 75% identical to the sequence of SEQ ID NO: 2. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 3. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 4. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 5. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 6. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence with SEQ ID NO: 7. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 8. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least is 75% identical to the sequence of SEQ ID NO: 9. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 10. In aspects of the present invention, the fusion protein includes an amino acid a sequence that is at least 75% identical to the sequence of SEQ ID NO: 11. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 12. In aspects of the present invention the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 13. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 14. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 75% identical to the sequence of SEQ ID NO: 15.

[0114] In embodiments of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 1. In aspects of the present invention, the fusion protein includes an amino acid sequence, which is at least 80% identical to the sequence of SEQ ID NO: 2. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 3. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 4. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 5. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 6. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 7. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 8. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least is 80% identical to the sequence of SEQ ID NO: 9. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 10. In aspects of the present invention, the fusion protein includes an amino acid a sequence that is at least 80% identical to the sequence of SEQ ID NO: 11. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 12. In aspects of the present invention the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 13. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 14. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 80% identical to the sequence of SEQ ID NO: 15.

[0115] In embodiments of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 1. In aspects of the present invention, the fusion protein includes an amino acid sequence, which is at least 85% identical to the sequence of SEQ ID NO: 2. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 3. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 4. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 5. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 6. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 7. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 8. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least is 85% identical to the sequence of SEQ ID NO: 9. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 10. In aspects of the present invention, the fusion protein includes an amino acid a sequence that is at least 85% identical to the sequence of SEQ ID NO: 11. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 12. In aspects of the present invention the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 13. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 14. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 85% identical to the sequence of SEQ ID NO: 15.

[0116] In embodiments of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 1. In aspects of the present invention, the fusion protein includes an amino acid sequence, which is at least 90% identical to the sequence of SEQ ID NO: 2. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 3. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 4. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 5. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 6. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence with SEQ ID NO: 7. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 8. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least is 90% identical to the sequence of SEQ ID NO: 9. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 10. In aspects of the present invention, the fusion protein includes an amino acid a sequence that is at least 90% identical to the sequence of SEQ ID NO: 11. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 12. In aspects of the present invention the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 13. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 14. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 90% identical to the sequence of SEQ ID NO: 15.

[0117] In embodiments of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 1. In aspects of the present invention, the fusion protein includes an amino acid sequence, which is at least 95% identical to the sequence of SEQ ID NO: 2. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 3. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 4. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 5. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 6. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 7. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 8. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least is 95% identical to the sequence of SEQ ID NO: 9. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 10. In aspects of the present invention, the fusion protein includes an amino acid a sequence that is at least 95% identical to the sequence of SEQ ID NO: 11. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 12. In aspects of the present invention the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 13. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 14. In aspects of the present invention, the fusion protein includes an amino acid sequence that is at least 95% identical to the sequence of SEQ ID NO: 15.

[0118] Complexes

[0119] In order for a fusion protein to perform epigenome editing, the fusion protein interacts with (eg, is non-covalently linked to) a polynucleotide (eg, segRNA) that is complementary to the target polynucleotide sequence (eg, the target DNA sequence to be edited) and further includes a sequence (ie, a binding sequence) to which a nuclease-deficient RNA-guided DNA endonuclease enzyme, as described herein, can bind. By forming such a complex, the fusion protein is appropriately positioned to perform epigenome editing. The term "complex" refers to a composition that includes two or more components, wherein the components bind together to form a functional unit. In aspects of the present invention, the complex described herein includes a fusion protein described herein and a polynucleotide described herein. Thus, in one aspect of the present invention there is provided a fusion protein as described herein, inclusive of its embodiments and aspects, and a polynucleotide comprising: (1) a DNA targeting sequence that is complementary to the target polynucleotide sequence; and (2) a binding sequence for a nuclease-deficient RNA-directed DNA endonuclease enzyme, wherein the nuclease-deficient RNA-directed DNA endonuclease enzyme is linked to the polynucleotide through a binding sequence (e.g., an amino acid sequence capable of binding to a DNA targeting sequence).

[0120] A DNA targeting sequence refers to a polynucleotide that includes a nucleotide sequence complementary to the target polynucleotide sequence (DNA or RNA). In aspects of the present invention, the DNA targeting sequence may be a single RNA molecule (single RNA polynucleotide), which may include a "single guide RNA" or "sgRNA". In aspects of the present invention, the DNA targeting sequence includes two RNA molecules (eg, joined together by hybridization at the site of a binding sequence (eg, at the site of a dCas9 binding sequence). In aspects of the present invention, the DNA targeting sequence (eg, egRNA) at at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, is 98% or 99% complementary to the target polynucleotide sequence. In aspects of the present invention, the DNA targeting sequence (e.g., sgRNA) is at least 50%, 55%, 60%, 65%, 70%, 75 %, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% complementary to a cellular gene sequence. In aspects of the present invention, the DNA targeting sequence (e.g., egRNA ) binds a cellular gene sequence. In aspects of the present invention, the DNA targeting sequence (eg, sRNA) is at least 75% complementary to the cellular gene sequence. In aspects of the present invention, the DNA targeting sequence (eg, sgRNA) is at least 80% complementary to the cellular gene sequence. In aspects of the present invention, the DNA targeting sequence (eg, sRNA) binds a cellular gene sequence. In aspects of the present invention, the DNA targeting sequence (eg, sgRNA) is at least 85% complementary to the cellular gene sequence. In aspects of the present invention, the DNA targeting sequence (eg, sRNA) binds a cellular gene sequence. In aspects of the present invention, the DNA targeting sequence (eg, sgRNA) is at least 90% complementary to the cellular gene sequence. In aspects of the present invention, the DNA targeting sequence (eg, sRNA) binds a cellular gene sequence. In aspects of the present invention, the DNA targeting sequence (eg, sgRNA) is at least 95% complementary to the cellular gene sequence. In aspects of the present invention, the DNA targeting sequence (eg, sRNA) binds a cellular gene sequence.

[0121] A "target polynucleotide sequence" as provided herein is a nucleic acid sequence that is present in or expressed by a cell for which a complementary targeting sequence (or DNA targeting sequence) is designed, wherein hybridization between the target sequence and guide sequence (or DNA targeting sequence) promotes the formation of the CRISPR complex. Full complementarity is not necessarily required as long as the complementarity is sufficient to induce hybridization and promote CRISPR complex formation. In aspects of the present invention, the target polynucleotide sequence is an exogenous nucleic acid sequence. In aspects of the present invention, the target polynucleotide sequence is an endogenous nucleic acid sequence.

[0122] The target polynucleotide sequence can be any region of a polynucleotide (eg, a DNA sequence) suitable for epigenome editing. In aspects of the present invention, the target polynucleotide sequence is part of a gene. In aspects of the present invention, the target polynucleotide sequence is part of a transcriptional regulatory sequence. In aspects of the present invention, the target polynucleotide sequence is part of a promoter, enhancer, or silencer. In aspects of the present invention, the target polynucleotide sequence is part of a promoter. In aspects of the present invention, the target polynucleotide sequence is part of an enhancer. In aspects of the present invention, the target polynucleotide sequence is part of a silencer.

[0123] In embodiments of the present invention, the target polynucleotide sequence is a hypomethylated nucleic acid sequence. The term “hypomethylated nucleic acid sequence” is used herein according to its standard meaning in the art and refers to the loss or absence of methyl groups on a 5-methylcytosine nucleotide (eg, a CpG). The loss or absence of methyl groups may be relative to the standard control. Hypomethylation may occur, for example, in aging cells or in cancer (eg, early stages of neoplasia) compared to a younger cell or non-cancerous cell, respectively. Thus, the complex may be useful in restoring normal (eg, immature or healthy) methylation levels.

[0124] In embodiments of the present invention, the target polynucleotide sequence is within about 3000 base pairs (bp) flanking the transcription start site. In aspects of the present invention, the target polynucleotide sequence is within about 3000, about 2900, about 2800, about 2700, about 2600, about 2500, about 2400, about 2300, about 2200, about 2100, about 2000, about 1900, about 1800, about 1700, about 1600, about 1500, about 1400, about 1300, about 1200, about 1100, about 1000, about 900, about 800, about 700, about 600, about 500, about 400, about 300, about 200 or about 100 pairs bases (bp) flanking the transcription start site.

[0125] In embodiments of the present invention, the target polynucleotide sequence is adjacent to, adjacent to, or within the promoter sequence. In aspects of the present invention, the target polynucleotide sequence is located within a CpG island. In aspects of the present invention, the target polynucleotide sequence is known to be associated with a disease or condition characterized by DNA hypomethylation.

[0126] In embodiments of the present invention, exemplary target polynucleotide sequences include those described in Tables 1 and 2. In aspects of the present invention, the target polynucleotide sequence includes the sequence of SEQ ID NO: 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93 or 95. In aspects of the present invention, the target polynucleotide sequence includes an amino acid sequence that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to SEQ ID NO: 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65 , 67, 69, 71, 73, 75, 77, 79, 81, 83, 85, 87, 89, 91, 93, or 95. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 37. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 39. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 41. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 43. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 45. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 47. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 49. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 51. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 53. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 55 In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 57. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 59. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 61. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 63. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 65. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: : 67. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 69. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 71. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 73. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 75. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 77. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 79. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 81. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 83. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 85. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 87. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 89. In aspects of the present invention, the target polynucleotide sequence is the sequence with SEQ ID NO: 91. In aspects of the present invention, the target polynucleotide sequence is the sequence with SEQ ID NO: 93. In aspects of the present invention, the target polynucleotide sequence is the sequence with SEQ ID NO: 95.

[0127] In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 37. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 39. In aspects of the present invention, the target polynucleotide sequence at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence of SEQ ID NO: 41. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93% , is 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 43. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 45. In aspects of the invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, is 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 47. In aspects of the invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, is 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 49. In aspects of the present invention, the target polynucleotide sequence is at least by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98% or by 99 % identical to the sequence of SEQ ID NO: 51. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94 %, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 53. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85 %, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 55. In aspects of the invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, is 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 57. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, is 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 59. In aspects of the present invention, the target polynucleotide sequence is at least by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98% or by 99 % identical to the sequence of SEQ ID NO: 61. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94 %, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 63. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85 %, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 65. In aspects of the invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, is 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 67. In aspects of the invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, is 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 69. In aspects of the present invention, the target polynucleotide sequence is at least by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98% or by 99 % identical to the sequence of SEQ ID NO: 71. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94 %, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 73. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85 %, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 75. In aspects of the invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, is 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 77. In aspects of the invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, is 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 79. In aspects of the present invention, the target polynucleotide sequence is at least by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98% or by 99 % identical to the sequence of SEQ ID NO: 81. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94 %, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 83. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85 %, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 85. In aspects of the invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, is 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 87. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, is 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 89. In aspects of the present invention, the target polynucleotide sequence is at least by 75%, by 80%, by 85%, by 90%, by 91%, by 92%, by 93%, by 94%, by 95%, by 96%, by 97%, by 98% or by 99 % identical to the sequence of SEQ ID NO: 91. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94 %, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 93. In aspects of the present invention, the target polynucleotide sequence is at least 75%, 80%, 85 %, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 95.

[0128] In aspects of this invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 37. In aspects of this invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 39. In aspects of this of the invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 41. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 43. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 45. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 47. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 49. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 51. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 53. In aspects of the present invention the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 55. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 57. In aspects of the present invention, the target polynucleotide sequence is at least is 90% identical to the sequence of SEQ ID NO: 59. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 61. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 63. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 65. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 67. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 69. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 71. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 71. the sequence is at least 90% identical to the sequence of SEQ ID NO: 73. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 75. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 75. % identical to the sequence of SEQ ID NO: 77. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 79. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 81. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 83. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 85. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 87. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 89. In aspects of the present invention, the target polynucleotide sequence of is at least 90% identical to the sequence of SEQ ID NO: 91. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical to the sequence of SEQ ID NO: 93. In aspects of the present invention, the target polynucleotide sequence is at least 90% identical sequences with SEQ ID NO: 95.

[0129] In aspects of this invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 37. In aspects of this invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 39. In aspects of this of the invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 41. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 43. In aspects of the present invention, the target polynucleotide sequence is the sequence of SEQ ID NO: 45. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 47. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 49. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 51. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 53. In aspects of the present invention the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 55. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 57. In aspects of the present invention, the target polynucleotide sequence is at least is 95% identical to the sequence of SEQ ID NO: 59. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 61. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 63. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 65. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 67. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 69. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 71. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 71. the sequence is at least 95% identical to the sequence of SEQ ID NO: 73. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 75. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 75. % identical to the sequence of SEQ ID NO: 77. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 79. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 81. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 83. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 85. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 87. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 89. In aspects of the present invention, the target polynucleotide sequence of is at least 95% identical to the sequence of SEQ ID NO: 91. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 93. In aspects of the present invention, the target polynucleotide sequence is at least 95% identical to the sequence of SEQ ID NO: 93. sequences with SEQ ID NO: 95.

[0130] In embodiments of the present invention, the complex includes dCas9 associated with a polynucleotide by binding to a polynucleotide binding sequence, thereby forming a ribonucleoprotein complex. In aspects of the present invention, the binding sequence forms a hairpin structure. In aspects of the present invention, the binding sequence is 30-100 nucleotides, 35-50 nucleotides, 37-47 nucleotides, or 42 nucleotides in length.

[0131] In embodiments of the present invention, a binding sequence (eg, a Cas9 binding sequence) interacts or binds to a Cas9 protein (eg, a dCas9 protein), and together they bind to a target polynucleotide sequence that is recognized by the DNA targeting sequence. A binding sequence (eg, a Cas9 binding sequence) includes two complementary stretches of nucleotides that hybridize to each other to form a double-stranded RNA duplex (dsRNA duplex). These two complementary stretches of nucleotides can be covalently linked through intermediate nucleotides known as linkers or linker nucleotides (for example, in the case of a single-molecule polynucleotide), and hybridize to form a double-stranded RNA duplex (dsRNA duplex, or "Cas9-binding hairpin") binding sequence ( for example, the Cas9-binding sequence), which leads to the formation of a stem-loop structure. Alternatively, in some aspects of the present invention, two complementary nucleotide regions may not be covalently linked, but instead are held together by hybridization between complementary sequences (eg, in the case of a two-molecule polynucleotide).

[0132] The binding sequence (e.g., a Cas9 binding sequence) may be from 10 nucleotides to 100 nucleotides in length, e.g., 10 nucleotides (nt) to 20 nt, 20 nt to 30 nt, 30 nt to 40 nt, 40 nt to 50 nt, from 50 nt to 60 nt, from 60 nt to 70 nt, from 70 nt to 80 nt, from 80 nt to 90 nt or from 90 nt to 100 nt. In aspects of the present invention, the binding sequence is between 15 nucleotides (nt) and 80 nt in length. In aspects of the present invention, the binding sequence is between 15 nt and 50 nt in length. In aspects of the present invention, the binding sequence is between 15 nt and 40 nt in length. In aspects of the present invention, the binding sequence is between 15 nt and 30 nt in length. In aspects of the present invention, the binding sequence is between 37 nt and 47 nt in length (eg, 42 nt). In aspects of the present invention, the binding sequence is between 15 nt and 25 nt in length.

[0133] The duplex of a dsRNA binding sequence (eg, a Cas9 binding sequence) can be from 6 base pairs (bp) to 50 bp in length. For example, a duplex of a dsRNA binding sequence (eg, a Cas9 binding sequence) can be as long as 6 bp. up to 40 bp, from 6 bp up to 30 bp, from 6 bp up to 25 bp, from 6 bp up to 20 bp, from 6 bp up to 15 bp, from 8 bp up to 40 bp, from 8 bp up to 30 bp, from 8 bp up to 25 bp, from 8 bp up to 20 p.o. or from 8 p.o. up to 15 p.o. In aspects of the present invention, the duplex dsRNA binding sequence (eg, Cas9 binding sequence) is between 8 bp in length. up to 10 p.o. In aspects of the present invention, the dsRNA duplex binding sequence (eg, Cas9 binding sequence) is between 10 bp in length. up to 15 p.o. In aspects of the present invention, the duplex dsRNA binding sequence (eg, Cas9 binding sequence) has a length of from 15 bp. up to 18 p.o. In aspects of the present invention, the duplex dsRNA binding sequence (eg, Cas9 binding sequence) has a length of from 18 bp. up to 20 p.o. In aspects of the present invention, the duplex dsRNA binding sequence (eg, Cas9 binding sequence) is between 20 bp in length. up to 25 p.o. In aspects of the present invention, the dsRNA duplex binding sequence (eg, Cas9 binding sequence) is between 25 bp in length. up to 30 p.o. In aspects of the present invention, the dsRNA duplex binding sequence (eg, Cas9 binding sequence) is between 30 bp in length. up to 35 p.o. In aspects of the present invention, the duplex dsRNA binding sequence (eg, Cas9 binding sequence) has a length of from 35 bp. up to 40 p.o. In aspects of the present invention, the duplex dsRNA binding sequence (eg, Cas9 binding sequence) is between 40 bp in length. up to 50 bp

[0134] In embodiments of the present invention, an exemplary polynucleotide that forms a complex with a fusion protein described herein includes those polynucleotides described in Tables 1 and 2 as sgRNA. In aspects of the present invention, the polynucleotide that forms a complex with the fusion protein described herein includes the sequence of SEQ ID NO: 38, 40, 42, 44, 46, 48, 50, 52, 54, 56, 58, 60 , 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92, or 94, or their corresponding RNA sequences. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes a sequence that is at least 80%, 85%, 90%, 91%, 92%, 93% , 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence with SEQ ID NO: 38, 40, 42, 44, 46, 48, 50, 52, 54, 56 , 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 82, 84, 86, 88, 90, 92 or 94, or their corresponding RNA sequences. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 38. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 40. In aspects of the present invention, the polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 42. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein, described herein includes the sequence of SEQ ID NO: 44. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 46. In aspects of the present invention, the polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 48. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: : 50. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 52. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 54. In aspects of the present invention, the polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 56. In aspects of the present invention, the polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 58. In aspects of the invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 60. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 62. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes a sequence with SEQ ID NO: 64. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 66. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described in herein includes the sequence of SEQ ID NO: 68. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 70. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 72. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 74 In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 76. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes includes the sequence of SEQ ID NO: 78. In aspects of the present invention, the polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 80. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 82. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 84. In aspects of the present invention a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 86. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 86. NO: 88. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 90. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein , includes the sequence of SEQ ID NO: 92. In aspects of the present invention, a polynucleotide that forms a complex with a fusion protein described herein includes the sequence of SEQ ID NO: 94.

[0135] Nucleic acids and vectors

[0136] The fusion protein described herein, including its embodiments and aspects, can be represented as a nucleic acid sequence encoding the fusion protein. Thus, in one aspect of the present invention there is provided a nucleic acid sequence encoding a fusion protein described herein, inclusive of its embodiments and aspects. In one aspect of the present invention, there is provided a nucleic acid sequence encoding a fusion protein described herein (including a DNA targeting sequence), inclusive of embodiments and aspects thereof. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein described herein, including fusion proteins having amino acid sequences with a certain % identity to the sequences described herein. In aspects of the present invention, the nucleic acid is RNA. In aspects of the present invention, the nucleic acid is messenger RNA. In aspects of the present invention, the messenger RNA is a messenger RNP. In aspects of the present invention, the nucleic acid sequence encodes the fusion proteins described herein, including their embodiments and aspects. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 1. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 2. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 3. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 4. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 5. In aspects of the present invention, the nucleic acid sequence encodes the fusion protein with the sequence SEQ ID NO: 6. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 7. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 8. In aspects of the present invention, the nucleic acid sequence acid encodes a fusion protein with the sequence SEQ ID NO: 9. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 10. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 11. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 12. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 13. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 13. 14. In aspects of the present invention, the nucleic acid sequence encodes a fusion protein with the sequence SEQ ID NO: 15.

[0137] It is further contemplated that a nucleic acid sequence encoding a fusion protein as described herein, inclusive of embodiments and aspects thereof, may be included in a vector. Thus, in one aspect of the present invention there is provided a vector including a nucleic acid sequence as described herein, inclusive of embodiments and aspects thereof. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein described herein, including fusion proteins having amino acid sequences with a certain % identity to the sequences described herein. In aspects of the present invention, the nucleic acid is messenger RNA. In aspects of the present invention, the messenger RNA is a messenger RNP. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 1. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 2. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 3. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 4. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 5. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 6. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 6. NO: 7. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 8. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 9. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 10. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 11. In aspects of the present invention, the vector contains a nucleic acid sequence acid that encodes a fusion protein with the sequence SEQ ID NO: 12. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 13. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes the fusion protein with the sequence SEQ ID NO: 14. In aspects of the present invention, the vector contains a nucleic acid sequence that encodes a fusion protein with the sequence SEQ ID NO: 15.

[0138] In embodiments of the present invention, the vector further includes a polynucleotide, wherein the polynucleotide includes: (1) a DNA targeting sequence that is complementary to the target polynucleotide sequence; and (2) a binding sequence for the enzyme nuclease-deficient RNA-guided DNA endonuclease. Thus, one or more vectors may include all the necessary components to perform epigenome editing.

[0139] Cells

[0140] The compositions described herein may be included in a cell. Within a cell, the compositions described herein, including their embodiments and aspects, can perform epigenome editing. Accordingly, in one aspect of the present invention there is provided a cell that includes a fusion protein as described herein, inclusive of embodiments and aspects thereof, a nucleic acid as described herein, inclusive of embodiments and aspects thereof, a complex, as described herein, inclusive of embodiments and aspects thereof, or a vector, as described herein, inclusive of embodiments and aspects thereof. In aspects of the present invention, there is provided a cell that includes a fusion protein as described herein, inclusive of embodiments and aspects thereof. In aspects of the present invention, there is provided a cell that includes a nucleic acid as described herein, inclusive of embodiments and aspects thereof. In aspects of the present invention, there is provided a cell that includes a complex as described herein, inclusive of embodiments and aspects thereof. In aspects of the present invention, there is provided a cell that includes a vector as described herein, inclusive of embodiments and aspects thereof. In aspects of the present invention, the cell is a eukaryotic cell. In aspects of the present invention, the cell is a mammalian cell.

[0141] Methods

[0142] It is contemplated that the compositions described herein can be used to edit the epigenome, and more specifically, to edit the epigenome resulting in the repression or silencing of target nucleic acid sequences (eg, genes). Without intending to be bound by theory, silencing may result from methylation and/or introduction of repressive chromatin markers (eg, mono-, di-, or trimethylation of certain histones (eg, H3K9, H3K27), deacetylation, acetylation, phosphorylation, ubiquitination) into chromatin containing the target nucleic acid sequence. Without intending to be bound by any theory, this method can be used to change the epigenetic state, for example, by closing chromatin through methylation or introducing repressive chromatin markers into chromatin containing a target nucleic acid sequence (eg, a gene). Without intending to be bound by any theory, it is proposed that the Dnmt3A-3L fusion protein functions to add methyl marks to DNA at CG sites found on CpG islands, and the KRAB domain recruits epigenetic factors that modify histones by introducing repressive marks. Without intending to be bound by any theory, DNA is methylated at the C nucleotide at CG sequences found on CpG islands (ie, adding methyl marks at the C nucleotide to DNA at CG sites found on CpG islands).

[0143] In one aspect of the present invention, there is provided a method for silencing a target nucleic acid sequence in a cell, including delivering a first polynucleotide encoding a fusion protein as described herein, inclusive of embodiments and aspects thereof, into a cell containing the target nucleic acid; and delivering into the cell a second polynucleotide that includes (i) a DNA targeting sequence that is complementary to the target nucleic acid sequence, and (ii) a binding sequence for the enzyme nuclease-deficient RNA-guided DNA endonuclease. Without intending to be bound by any theory, the fusion protein causes silencing of a target nucleic acid sequence in a cell by methylating the chromatin containing the target nucleic acid sequence and/or by introducing repressive chromatin markers into the chromatin containing the target nucleic acid sequence. Without intending to be bound by any theory, chromatin methylation means that DNA is methylated at the C nucleotide at CG sequences found on CpG islands (ie, adding methyl marks at the C nucleotide to DNA at CG sites found on CpG islands). In aspects of the present invention, methylated is a sequence that is within about 3000 base pairs of the target nucleic acid sequence. In aspects of the invention, a sequence that is methylated is within about 3000, about 2900, about 2800, about 2700, about 2600, about 2500, about 2400, about 2300, about 2200, about 2100, about 2000, about 1900, about 1800 , about 1700, about 1600, about 1500, about 1400, about 1300, about 1200, about 1100, about 1000, about 900, about 800, about 700, about 600, about 500, about 400, about 300, about 200 or about 100 base pairs from the target nucleic acid sequence.

[0144] The term “repressive chromatin markers” as used herein refers to modifications made to chromatin that result in silencing (eg, reduction or inhibition of transcription) of a target nucleic acid sequence (eg, a gene). Examples of repressive chromatin markers include, but are not limited to, histone mono-, di- and/or trimethylation, acetylation/deacetylation, phosphorylation and ubiquitination (eg, H3K9, H3K27, H3K79, H2BK5).

[0145] In embodiments of the present invention, silencing refers to the complete suppression of transcription. In aspects of the present invention, silencing refers to a significant reduction in transcription compared to control levels of transcription.

[0146] In embodiments of the present invention, the first polynucleotide is contained in the first vector. In aspects of the present invention, the first polynucleotide is contained in a second vector. In aspects of the present invention, the first vector and the second vector are the same. In aspects of the present invention, the first vector is different from the second vector.

[0147] In embodiments of the present invention, a polynucleotide described herein is delivered into a cell by any method known in the art, for example, by transfection, electroporation, or transduction.

[0148] Alternatively, in one aspect of the present invention, a method is provided that results in silencing of a target nucleic acid sequence in a cell, including delivery of a complex as described herein, inclusive of embodiments and aspects thereof, into a cell that contains the target nucleic acid. Without intending to be bound by any theory, the complex causes silencing of a target nucleic acid sequence in a cell by methylating the chromatin containing the target nucleic acid sequence and/or by introducing repressive chromatin markers into the chromatin containing the target nucleic acid sequence.

[0149] In embodiments of the present invention, the cell is a mammalian cell.

[0150] In embodiments of the present invention, the method has a specificity that is 2 times higher than the specificity for a non-target nucleic acid sequence. In aspects of the invention, the method has a specificity that is at least 2-fold (e.g., 2-fold, 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 15-fold, 20-fold). , 25 times) higher than the specificity for the non-target nucleic acid sequence. Methods for determining specificity are well known in the art and include, but are not limited to, RNA sequencing, bisulfite sequencing, chromatin immunoprecipitation, flow cytometry, and quantitative PCR. Thus, in aspects of the present invention, specificity is determined using an RNA sequencing method. In aspects of the present invention, specificity is determined using a bisulfite sequencing method. In aspects of the present invention, specificity is determined using a chromatin immunoprecipitation method. In aspects of the present invention, specificity is determined using flow cytometry. In aspects of the present invention, specificity is determined using a quantitative PCR method.

[0151] In aspects of the present invention, the complex is delivered into the cell by any means known in the art, for example, through the delivery of ribonucleoprotein (RNP).

[0152] Embodiments N1 - N41

[0153] Embodiment of the invention N1. Fusion protein containing the enzyme nuclease-deficient RNA-guided DNA endonuclease box-linked domain , and a DNA methyltransferase domain.

[0154] Embodiment of the invention N2. The fusion protein of embodiment N1, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9, ddCpf1, a nuclease-deficient variant of Cas9, or a class II nuclease-deficient CRISPR endonuclease.

[0155] Embodiment of the invention N3. The fusion protein of embodiment N1 or N2, wherein said DNA methyltransferase domain is a Dnmt3A-3L domain.

[0156] Embodiment of the invention N4. The fusion protein of embodiment N1, wherein the fusion protein includes, from N-terminus to C-terminus, a DNA methyltransferase domain, an enzyme-deficient RNA-guided DNA endonuclease, and a box-linked domain .

[0157] Embodiment of the invention N5. The fusion protein of embodiment N4, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9 and said DNA methyltransferase domain is a Dnmt3A-3L domain.

[0158] Embodiment of the invention N6. The fusion protein of embodiment N5, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9 and said DNA methyltransferase domain is a Dnmt3A-3L domain.

[0159] Embodiment of the invention N7. The fusion protein of embodiment N6, wherein said peptide linker is an XTEN linker.

[0160] Embodiment of the invention N8. The fusion protein of embodiment N1, wherein the fusion protein includes, in the N-terminal to C-terminal direction, a box-linked domain , a nuclease-deficient RNA-guided DNA endonuclease enzyme and a DNA methyltransferase domain.

[0161] Embodiment of the invention N9. The fusion protein of embodiment N8, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9 and said DNA methyltransferase domain is a Dnmt3A-3L domain.

[0162] Embodiment of the invention N10. The fusion protein of embodiment N9, wherein said dCas9 is covalently linked to the Dnmt3A-3L domain via a peptide linker, and wherein the domain is associated with the box , is covalently linked to dCas9 through a peptide linker.

[0163] Embodiment of the invention N11. The fusion protein of embodiment N10, wherein said peptide linker is an XTEN linker.

[0164] Embodiment of the invention N12. The fusion protein of any one of embodiments N1 to N3, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is covalently linked to a box-linked domain , via a peptide linker.

[0165] Embodiment of the invention N13. The fusion protein of any one of embodiments N1 to N3, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is covalently linked to a DNA methyltransferase domain through a peptide linker.

[0166] Embodiment of the invention N14. The fusion protein of any one of embodiments N1 to N3, characterized in that said box-linked domain , is covalently linked to the DNA methyltransferase domain via a peptide linker.

[0167] Embodiment of the invention N15. The fusion protein of any one of embodiments N12 to N14, wherein said peptide linker is an XTEN linker.

[0168] Embodiment of the invention N16. The fusion protein of embodiment N15, wherein said XTEN linker contains from about 16 to about 80 amino acid residues.

[0169] Embodiment of the invention N17. The fusion protein of any one of embodiments N1 to N16, further comprising a nuclear localization signal peptide.

[0170] Embodiment of the invention N18. The fusion protein of embodiment N1, wherein said fusion protein contains an amino acid sequence with SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15.

[0171] Embodiment of the invention N19. A nucleic acid sequence encoding a fusion protein according to any one of embodiments N1 to N18.

[0172] Embodiment of invention N20. The nucleic acid sequence according to embodiment N19, characterized in that said nucleic acid sequence is messenger RNA.

[0173] Embodiment of invention N21. A complex comprising: (i) a fusion protein according to any one of embodiments N1 to N18; and (ii) a polynucleotide comprising: (a) a DNA targeting sequence complementary to the target polynucleotide sequence; and (b) a binding sequence for a nuclease-deficient RNA-guided DNA endonuclease enzyme, wherein the nuclease-deficient RNA-guided DNA endonuclease enzyme is linked to the polynucleotide through a binding sequence.

[0174] Embodiment of invention N22. The complex according to embodiment N21, characterized in that said target polynucleotide sequence is part of a gene.

[0175] Embodiment of invention N23. The complex according to embodiment N21, characterized in that said target polynucleotide sequence is part of a transcription regulatory sequence.

[0176] Embodiment of invention N24. The complex according to embodiment N21, characterized in that said target polynucleotide sequence is part of a promoter, enhancer or silencer.

[0177] Embodiment of invention N25. The complex according to embodiment N21, characterized in that said target polynucleotide sequence is within about 3000 bp. and flanks the transcription start site.

[0178] Embodiment of invention N26. A vector containing a nucleic acid sequence according to embodiment N19 or N20.

[0179] Embodiment of invention N27. The vector of embodiment N26, further comprising a polynucleotide, wherein the polynucleotide contains: (a) a DNA targeting sequence that is complementary to the target polynucleotide sequence; and (b) a binding sequence for the enzyme nuclease-deficient RNA-guided DNA endonuclease.

[0180] Embodiment of invention N28. A cell containing a fusion protein according to any one of embodiments N1 to N18; nucleic acid according to embodiment N19 or N20; the complex of any one of embodiments N21 to N25, or the vector of embodiment N26 or N27.

[0181] Embodiment of invention N29. A cell according to embodiment N28, characterized in that said cell is a eukaryotic cell.

[0182] Embodiment of the invention N30. A cell according to embodiment N28, characterized in that said cell is a mammalian cell.

[0183] Embodiment of the invention N31. A method of silencing a target nucleic acid sequence in a cell, comprising: (i) delivering a first polynucleotide encoding the fusion protein of any one of embodiments N1 to N18 into a cell containing the target nucleic acid; and (ii) delivering into the cell a second polynucleotide that includes (a) a DNA targeting sequence that is complementary to the target nucleic acid sequence, and (b) a binding sequence for the enzyme nuclease-deficient RNA-guided DNA endonuclease.

[0184] Embodiment of invention N32. The method according to embodiment N31, characterized in that said fusion protein causes silencing of a target nucleic acid sequence in a cell by methylating the chromatin containing the target nucleic acid sequence and/or by introducing repressive chromatin markers into the chromatin containing the target nucleic acid sequence.

[0185] Embodiment of invention N33. The method according to embodiment N31 or N32, characterized in that said first polynucleotide is contained in a first vector.

[0186] Embodiment of invention N34. The method according to any one of embodiments N31 to N33, characterized in that said first polynucleotide is contained in a second vector.

[0187] Embodiment of the invention N35. The method according to embodiment N34, characterized in that said first vector and second vector are the same.

[0188] Embodiment of invention N36. The method according to embodiment N34, characterized in that said first vector is different from the second vector.

[0189] Embodiment of invention N37. The method according to embodiment N31, characterized in that said cell is a mammalian cell.

[0190] Embodiment of invention N38. The method according to embodiment N31, characterized in that the specificity of this method is 2 times higher than the specificity with respect to the sequence of the non-target nucleic acid.

[0191] Embodiment of invention N39. A method for silencing a target nucleic acid sequence in a cell, comprising delivering the complex according to any of embodiments N21 to N25 into a cell containing the target nucleic acid.

[0192] Embodiment of the invention N40. The method according to embodiment N39, characterized in that the complex causes silencing of the target nucleic acid sequence in the cell by methylation of the chromatin containing the target nucleic acid sequence and (or) by introducing repressive chromatin markers into the chromatin containing the target nucleic acid sequence.

[0193] Embodiment of the invention N41. The method according to embodiment N39 or N40, characterized in that said cell is a mammalian cell.

[0194] Embodiment of the invention N42. The method according to any of the embodiments from N39 to N41, characterized in that the specificity of this method is 2 times higher than the specificity with respect to the sequence of the non-target nucleic acid.

[0195] Embodiments 1 to 36

[0196] Embodiment 1. A fusion protein containing a nuclease-deficient RNA-guided DNA endonuclease enzyme box-linked domain (KRAB domain), and DNA methyltransferase domain.

[0197] Embodiment 2. The fusion protein of Embodiment 1, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9, ddCpf1, a nuclease-deficient Cas9 variant, or a class II nuclease-deficient CRISPR endonuclease.

[0198] Embodiment 3. The fusion protein of Embodiment 1 or 2, wherein said DNA methyltransferase domain is a Dnmt3A-3L domain.

[0199] Embodiment 4. The fusion protein of any one of Embodiments 1 to 3, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is covalently linked to a KRAB domain via a peptide linker.

[0200] Embodiment 5. The fusion protein of any one of Embodiments 1 to 4, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is covalently linked to a DNA methyltransferase domain through a peptide linker.

[0201] Embodiment 6. The fusion protein of any one of Embodiments 1 to 5, wherein said KRAB domain is covalently linked to a DNA methyltransferase domain through a peptide linker.

[0202] Embodiment 7. The fusion protein of any one of Embodiments 4 to 6, wherein said peptide linker is an XTEN linker.

[0203] Embodiment 8. The fusion protein of Embodiment 7, wherein said XTEN linker contains from about 16 to about 80 amino acid residues.

[0204] Embodiment 9. The fusion protein of any one of Embodiments 1 to 8, further comprising a nuclear localization signal peptide.

[0205] Embodiment 10. The fusion protein of any one of Embodiments 1 to 9, wherein said fusion protein contains, in an N-terminal to C-terminal direction, a KRAB domain, an RNA nuclease-deficient enzyme. a directed DNA endonuclease, and a DNA methyltransferase domain.

[0206] Embodiment 11. The fusion protein of any one of Embodiments 1 to 10, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9 and said DNA methyltransferase domain is a Dnmt3A-3L domain. .

[0207] Embodiment 12. The fusion protein of Embodiment 11, wherein the dCas9 is covalently linked to the KRAB domain via a peptide linker, and wherein the dCas9 is covalently linked to the Dnmt3A-3L domain via a peptide linker.

[0208] Embodiment 13. The fusion protein of Embodiment 12, wherein said peptide linker is an XTEN linker.

[0209] Embodiment 14. The fusion protein of any one of Embodiments 1 to 13, wherein said fusion protein contains the amino acid sequence of SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 12, 13, 14 or 15.

[0210] Embodiment 15. A nucleic acid sequence encoding the fusion protein of any one of Embodiments 1 to 14.

[0211] Embodiment 16. A complex comprising: (i) the fusion protein of any one of Embodiments 1 to 18; and (ii) a polynucleotide comprising: (a) a DNA targeting sequence complementary to the target polynucleotide sequence; and (b) a binding sequence for a nuclease-deficient RNA-guided DNA endonuclease enzyme, wherein the nuclease-deficient RNA-guided DNA endonuclease enzyme is linked to the polynucleotide through a binding sequence.

[0212] Embodiment 17. The complex of Embodiment 21, wherein said target polynucleotide sequence is part of a gene.

[0213] Embodiment 18. The complex of Embodiment 21, wherein said target polynucleotide sequence is part of a transcription regulatory sequence.

[0214] Embodiment 19. The complex of Embodiment 21, wherein said target polynucleotide sequence is part of a promoter, enhancer, or silencer.

[0215] Embodiment 20. The complex of Embodiment 21, wherein said target polynucleotide sequence is a hypomethylated nucleic acid sequence.

[0216] Embodiment 21. The complex of Embodiment 21, wherein said target polynucleotide sequence is within about 3000 bp. and flanks the transcription start site.

[0217] Embodiment 22. A vector containing the nucleic acid sequence of Embodiment 19.

[0218] Embodiment 23. The vector of Embodiment 26, further comprising a polynucleotide, wherein the polynucleotide comprises: (a) a DNA targeting sequence that is complementary to the target polynucleotide sequence; and (b) a binding sequence for the enzyme nuclease-deficient RNA-guided DNA endonuclease.

[0219] Embodiment 24. A cell containing the fusion protein of any one of Embodiments 1 to 14; nucleic acid according to embodiment 15; complex according to any of the embodiments of the invention from 16 to 21; or a vector according to embodiment 22 or 23.

[0220] Embodiment 25. The cell of Embodiment 28, wherein said cell is a eukaryotic cell.

[0221] Embodiment 26. The cell of Embodiment 28, wherein said cell is a mammalian cell.

[0222] Embodiment 27. A method for silencing a target nucleic acid sequence in a cell, comprising: (i) delivering a first polynucleotide encoding the fusion protein of any one of Embodiments 1 to 14 to a cell containing the target nucleic acid; and (ii) delivering into the cell a second polynucleotide that includes (a) a DNA targeting sequence that is complementary to the target nucleic acid sequence, and (b) a binding sequence for the enzyme nuclease-deficient RNA-guided DNA endonuclease, wherein said fusion the protein causes silencing of a target nucleic acid sequence in a cell by methylating the chromatin containing the target nucleic acid sequence and/or by introducing repressive chromatin markers into the chromatin containing the target nucleic acid sequence.

[0223] Embodiment 28. The method of Embodiment 27, wherein said first polynucleotide is contained in a first vector.

[0224] Embodiment 29. The method of Embodiment 27, wherein said first polynucleotide is contained in a second vector.

[0225] Embodiment 30. The method of Embodiment 28 or 29, wherein said first vector and second vector are the same.

[0226] Embodiment 31. The method of Embodiment 28 or 29, wherein said first vector is different from the second vector.

[0227] Embodiment 32. The method of any one of Embodiments 27 to 31, wherein the cell is a mammalian cell.

[0228] Embodiment 33. The method according to any one of embodiments 27 to 32, wherein the specificity of the method is 2 times higher than the specificity with respect to the non-target nucleic acid sequence.

[0229] Embodiment 34. A method for silencing a target nucleic acid sequence in a cell, comprising delivering the complex of any one of embodiments 16 to 20 to a cell containing the target nucleic acid, wherein the complex causes silencing of the target nucleic acid sequence in cell by: (i) methylating the chromatin containing the target nucleic acid sequence, (ii) introducing repressive chromatin markers into the chromatin containing the target nucleic acid sequence, or (iii) methylating the chromatin containing the target nucleic acid sequence and introducing repressive chromatin markers into chromatin containing the target nucleic acid sequence.

[0230] Embodiment 35. The method of Embodiment 34, wherein said cell is a mammalian cell.

[0231] Embodiment 36. The method according to embodiment 34 or 35, wherein the specificity of the method is 2 times higher than the specificity with respect to the non-target nucleic acid sequence.

EXAMPLES

[0232] Embodiments of the invention and aspects set forth herein are further illustrated by the following examples. The examples are intended to illustrate embodiments and aspects of the invention only, and should not be construed as limiting the scope of the invention set forth herein.

[0233] Example 1

[0234] Epigenetic modulators fused to dCas9 have been tested for persistent gene silencing. The original version (B1, p76 (SEQ ID NO: 1)) of the all-in-one protein (Fig. 1A) contains a KRAB domain fused to the -N terminus of dCas9 (SEQ ID NO: 23), separated by a GGSGGGS linker (SEQ ID NO: 17), and Dnmt3A - Dnmt3L at the C terminus of dCas9 (separated by the linker EASGSGRASPGIPGSTR (SEQ ID NO: 19)). Another all-in-one protein that combined the domain KRAB (SEQ ID NO: 16), dCas9 (D10A, H208A), Dnmt3A - Dnmt3L (SEQ ID NO: 33; where SEQ ID NO: 26 is Dnmt3A and SEQ ID NO: 28 is Dnmt3L) into a single polypeptide (Figure 1B). With reference to FIG. 1B, the dCas9 - KRAB protein was adapted from Gilbert et al., Cell 2013, for use with CRISPR interference (CRISPRi) technology, and the dCas9 - Dnmt3A - Dnmt3L fusion protein was adapted from Stepper et al., Nucleic Acids Research, 2016.

[0235] Epigenetic editor B1 activity was tested in HEK293T cells using a DNA methylation-sensitive GFP reporter (adapted from Stelzer et al., Cell 2015) to assess long-term silencing caused by the all-in-one protein (Figure 1C) . A universal chromatin opening element (UCOE) was added upstream of the GAPDH CpG island (CGI) to prevent background silencing of the lentiviral vector in mammalian cells. The gfp gene is switched off when GAPDH CGI is methylated. A, B, and C indicate the positions that we encoded for single guide RNA (sgRNA) targeting in the promoter. These target sequences and the corresponding egRNA sequences are listed in Table 1 below. Two plasmids were cotransfected into the cells, one encoding the hit-and-run mutant protein and the other plasmid encoding the egRNA (Fig. 1D). Two days after transfection, cells that express the hit-and-run protein and a vector expressing egRNA were sorted. GFP fluorescence was assessed over time using flow cytometry. A population of cells undergoing long-term silencing of the GFP reporter was observed when the all-in-one protein was expressed with egRNA ( Fig. 1E ). The number of cells undergoing long-term silencing was higher than when exposed to dCas9 - Dnmt3A - Dnmt3L (without the KRAB domain).

[0236] Table 1

Name Target sequence (5' to 3') egRNA sequence (5' to 3') A (JKNg156) ACTGCGGAAATTTGAGCGT
(SEQ ID NO: 37) ACGCTCAAATTTCCGCAGT
(SEQ ID NO: 38) B (JKNg158) AGGCAATGGCTGCACATGC
(SEQ ID NO: 39) GCATGTGCAGCCATTGCCT
(SEQ ID NO: 40) C (JKNg160) GACGCTTGGTTCTGAGGAG
(SEQ ID NO: 41) CTCCTCAGAACCAAGCGTC
(SEQ ID NO: 42)

[0237]Silencing The GFP reporter is egRNA sequence dependent, with guide sequence C causing the highest level of silencing among the three egRNA sequences tested. Combining egRNAs encoding different sequences into a single pool did not lead to a significant change in gene silencing.

[0238] Example 2

[0239] Three genes (CD29, CD81, CD151) were targets for long-term silencing using a hit-and-run fusion protein. All three proteins are localized to the cell surface, and knockdown was assessed by staining cells with cell surface antibodies followed by flow cytometry. Representative flow cytometry data obtained 22 days after transfection are shown in Fig. 2A - 2C. Quadrant IV represents cells that have turned off the gene, as indicated by the percentage of cells with the gene turned off. The absence of cells in quadrants I and II means that the hit-and-run protein (containing the SFB tag) is no longer present in the cells. In FIG. Figure 2D shows a quantitative assessment of the silencing of the CD29, CD81 and CD151 genes by three different egRNAs, caused by three different egRNA sequences or a pool of all three egRNAs. The target DNA sequences and their sgRNAs used in this experiment are summarized in Table 2.

[0240] Table 2

Name Target sequence (5' to 3') egRNA sequence (5' to 3') CD29, sgRNA - A TCCGGAAACGCATTCCTCT
(SEQ ID NO: 43) AGAGGAATCGGTTTCCGGA
(SEQ ID NO: 44) CD29, egRNA - B CCGCGTCAGCCCGGCCCGG (SEQ ID NO: 45) CCGGGCCGGGCTGACGCGG
(SEQ ID NO: 46) CD29, egRNA - C CGACTCCCGCTGGGCCTCT (SEQ ID NO: 47) AGAGGGCCCAGCGGGAGTCG
(SEQ ID NO: 48) CD81, sgRNA - A ccgttgcgcgctcgctctc
(SEQ ID NO: 49) gagagcgagcgcgcaacgg
(SEQ ID NO: 50) CD81, sgRNA - B CCGCGCATCCTGCCAGGCC (SEQ ID NO: 51) GGCCTGGCAGGATGCGCGG
(SEQ ID NO: 52) CD81, egRNA - C CCAACTTGGCGCGTTTCGG (SEQ ID NO: 53) CCGAAACGCGCCAAGTTGG
(SEQ ID NO: 54) CD151, sgRNA - A ACCACGCGTCCGAGTCCGG (SEQ ID NO: 55) CCGGACTCGGACGCGTGGT
(SEQ ID NO: 56) CD151, sgRNA - B TGCTCATTGTCCCTGGACA
(SEQ ID NO: 57) TGTCCAGGGACAATGAGCA
(SEQ ID NO: 58) CD151, sgRNA - C GGACACCCTGCTCATTGTC
(SEQ ID NO: 59) GACAATGAGCAGGGTGTCC
(SEQ ID NO: 60)

[0241] Two or three genes were targeted simultaneously to show that an all-in-one protein could be multiplexed by co-delivering egRNAs targeting different genes. egRNA NC refers to the non-targeting control of egRNA. The results are shown in Fig. 2E.

[0242] Gene silencing was observed in cells that originated as a single clone, and in the majority of cells the target CLTA gene remained switched off (37 of 39 clones). Graph in Fig. 2F represents a time point taken 9 months after transfection of the all-in-one protein and egRNA targeting the CLTA gene.

[0243] The system described herein can target any genes in mammalian genomes, especially those that contain CpG islands at the gene promoter. Dnmt3A – Dnmt3L canonically targets CpG dinucleotides. Examples of genes that can be targeted by this system include, but are not limited to, CXCR4, CD4, CD8, CD45, PD-1, CLTA-4, TGFBR, TCRa, TCRb, B2M.

[0244] Example 3

[0245] Cells that had lost expression of ITGB1 (CD29), CD81, and CD151 thirty-six days after transfection were collected and their RNA expression profiles were analyzed. As shown in FIG. 3A - 3C, successful knockdown of target genes was found compared to the untargeted egRNA control. Fig. 3D - 3F are volcano plots showing that target gene knockdown is the only significant gene knockdown for each experiment, indicating the high specificity of gene silencing. Fig. 3G to 3I are quantifications of transcript levels showing greater than 96% knockdown of the target gene.

[0246] Example 4

[0247] The all-in-one protein can be transfected and expressed in HeLa (cervical cells), U2OS (bone cells), and human induced pluripotent stem cells (iPSCs). Flow cytometry plots in Fig. 4A - 4F show expression of SFB, which is fused to the protein. The targets were three endogenous genes in HeLa and U2OS cells (i.e., CD29, CD81, and CD151). As shown in FIG. 4G, stable silencing was detected, measured 18 days after transfection. By targeting Pcsk9, Npc1, Spcs1 and Cd81, gene silencing was detected in AML12 mouse hepatocyte cell lines. Silencing was detected by qPCR 14 days after transfection, as shown in Fig. 4H. The sgRNA sequences used in this experiment are given in Table 3.

[0248] Table 3

Name Target sequence
(from 5' to 3') egRNA sequence (5' to 3') Pcsk9 egRNA - 1 TCCGGAAACGCATTCCTCT
(SEQ ID NO: 43) AGAGGAATCGGTTTCCGGA
(SEQ ID NO: 44) Pcsk9 egRNA - 2 ACCGGCAGCCTGCGCGTCC
(SEQ ID NO: 61) GGACGCGCAGGCTGCCCGGT
(SEQ ID NO: 62) Pcsk9 egRNA - 3 CGATGGGCACCCACTGCTC
(SEQ ID NO: 63) GAGCAGTGGGTGCCCATCG
(SEQ ID NO: 64) Pcsk9 egRNA - 4 CCTTCACGTGGACGCGCAG
(SEQ ID NO: 65) CTGCGCGTCCACGTGAAGG
(SEQ ID NO: 66) Pcsk9 egRNA - 5 CGTGAAGGTGGAAGCCTTC
(SEQ ID NO: 67) GAAGGCTTCCACCTTCACG
(SEQ ID NO: 68) Npc1 egRNA - 1 CTCCTTGGTCAGGCGCCCGG
(SEQ ID NO: 69) CCGGCGCCTGACCAAGGAG
(SEQ ID NO: 70) Npc1 egRNA - 2 TGGTCAGGCGCGGTTCCG
(SEQ ID NO: 71) CGGAACCGGCGCCTGACCA
(SEQ ID NO: 72) Npc1 egRNA - 3 TAGAGGTCGCTTCTCCTC
(SEQ ID NO: 73) GAGGAGAAGGCGACCTCTA
(SEQ ID NO: 74) Npc1 egRNA - 4 CGACGCTCGGGTCGCGGTG
(SEQ ID NO: 75) CACCGCGACCCGAGCGTCG
(SEQ ID NO: 76) Npc1 egRNA - 5 ATGCTGTCGCCGCGCGGGG
(SEQ ID NO: 77) CCCGCGGCGACAGCAT
(SEQ ID NO: 78) Spcs1 egRNA - 1 CTCACCCTCACCGGAGCCA
(SEQ ID NO: 79) TGGCTCCGGTGAGGGTGAG
(SEQ ID NO: 80) Spcs1 egRNA - 2 CCGCAAACTTTACTCCTTA
(SEQ ID NO: 81) TAAGGAGTAAAGTTTGCGG
(SEQ ID NO: 82) Spcs1 egRNA - 3 CTCGGAGACATCCGCTTCC
(SEQ ID NO: 60) GGAAGCGGATGTCTCCGAG
(SEQ ID NO: 60) Spcs1 egRNA - 4 CTCCTAAGATTGGCTTCAC
(SEQ ID NO: 83) GTGAAGCCAATCTAGGAG
(SEQ ID NO: 84) Spcs1 egRNA - 5 CCGGAGCCACTCCTAAGAT
(SEQ ID NO: 85) ATCTTAGGAGTGGCTCCGG
(SEQ ID NO: 86) Cd81 egRNA - 1 TTCTCTACCTACGTCTCA
(SEQ ID NO: 87) TGAGACGTAGGGTAGAGAA
(SEQ ID NO: 88) Cd81 egRNA - 2 TACGTCTCATTCTCCGCAA
(SEQ ID NO: 89) TTGCGGAGAATGAGACGTA
(SEQ ID NO: 90) Cd81 egRNA - 3 GCTAGGCCTCCAGCCCTTC
(SEQ ID NO: 91) GAAGGGCTGGAGGCCTAGC
(SEQ ID NO: 92) Cd81 egRNA - 4 ACAGGTGGCGCCGCAACTT
(SEQ ID NO: 93) AAGTTGCGGCGCCACCTGT
(SEQ ID NO: 94) Cd81 egRNA - 5 AGCCGGAGGCGCGAGAGTC
(SEQ ID NO: 95) GACTCTCGCGCCTCCGGCT
(SEQ ID NO: 96)

[0249] Example 5

[0250] In FIG. Figure 5 shows a diagram of the all-in-one protein constructs that were designed and tested for gene silencing. The original format (p76, B1) SEQ ID NO: 1 has been modified to encode XTEN linkers (e.g., 16 amino acids (SEQ ID NO: 31) or 80 amino acids (SEQ ID NO: 32)) at either the N-terminus or C -end of dCas9 (SEQ ID NO: 29). All vectors contain HA tags (SEQ ID NO: 24) at the C terminus of dCas9. Aspects of the present invention use the CAG promoter because it provides good expression, for example, in the p76 and p90-102, p112 (B2) constructs. With reference to FIG. 5, protein constructs p90 to p102 correspond to SEQ ID NO: 2-14, respectively, and protein constructs p112 correspond to SEQ ID NO: 15.

[0251] Example 6

[0252] The protein constructs shown in FIG. 5 were tested for silencing of the CLTA gene in HEK293T cells for 18 days after transfection (Fig. 6A - 6B). Variable levels of gene silencing activity were found, including a number of variants with more robust gene silencing compared to the p76 (B1) construct, such as p99 (SEQ ID NO: 11), p100 (SEQ ID NO: 12) and p112 (SEQ ID NO: 15). In FIG. 6A and 6B show that the dCas9 - KRAB and dCas9 - Dnmt3A - Dnmt3L constructs demonstrated short-term and lower long-term silencing efficiencies.

[0253] p76 (SEQ ID NO: 1), p112 (SEQ ID NO: 15) were tested for silencing of the endogenous HIST2H2BE (H2B) gene and the synthetic reporter gene Snrpn-GFP stably expressed in HEK293T cells (Fig. 6C - 6D) . The cells were observed for 50 days after transfection. The p112 variant supported gene silencing with higher efficiency than the p76 construct (B1). The dCas9 - Dnmt3A - Dnmt3L and dCas9 - KRAB fusion proteins showed short-term and lower long-term silencing efficiency. In FIG. Figure 6E shows a graph of p76 and p112 protein expression over 50 days to turn off the HIST2H2BE (H2B) gene. Protein levels were measured using a flow cytometry method to detect SBF, which is co-expressed with the all-in-one protein.

[0254] Example 7

[0255] Western blot analysis was performed on all-in-one protein variants p76, p90 - p102 using the anti- Steptococcus pyogenes Cas9 antibody. With reference to FIG. 7A, the top band represents the full-length protein and the smaller bands represent the all-in-one protein proteolysis products. Variants that exhibit weak proteolysis, such as p99 (SEQ ID NO: 11), p100 (SEQ ID NO: 12) and p102 (SEQ ID NO: 14), show higher efficiency of gene silencing. Variants with high levels of proteolysis, such as p96 (SEQ ID NO: 8) and p97 (SEQ ID NO: 9), resulted in lower efficiency of sustained gene silencing.

[0256] Western blot analysis was performed with all-in-one protein variants to detect free Dnmt3A that is cleaved from the fusion protein. As shown in FIG. 7B, variants for which little or no free Dnmt3 was detected, such as p92 (SEQ ID NO: 4), p100 (SEQ ID NO: 12), p101 (SEQ ID NO: 13) and p102 (SEQ ID NO: : 14), exhibited a higher efficiency of stable gene silencing compared to variants with detectable cleaved Dnmt3A, i.e. p76 (SEQ ID NO: 1), p91 (SEQ ID NO: 3), p96 (SEQ ID NO: 8), p98 (SEQ ID NO: 10).

[0257] Example 8

[0258] A pooled screening was performed as shown in FIG. 8A, with the aim of identifying optimal egRNAs that lead to long-term gene silencing. Four HEK293T cell lines were used, each with a different GFP-tagged gene (CLTA, VIM, HIST2H2BE (H2B), and RAB11A). Tiling libraries consisting of egRNAs spanning +/- 2.5 kb. from the transcription start site (TST) of each gene, were stably expressed in cells by delivery of lentiviruses followed by transient expression of plasmid DNA expressing the all-in-one protein. Four weeks after transfection, cells that supported gene silencing were sorted to identify specific egRNAs. Fig. 8B - 8E are flow cytometry histograms showing the percentage of cells undergoing gene silencing four weeks after transfection.

[0259] FIG. 9A - 9D are maps of egRNA functionality at the transcription start site of the target gene (CLTA, H2B, RAB11, VIM). The transcription start site (TSS) and CpG island are annotated above each graph. Each dot represents one egRNA, and its effectiveness in long-term gene silencing is displayed as a change in log2-fold egRNA content. The degree of nucleosome occupancy (lower graph) is plotted against the signal from MNase (micrococcal nuclease, English “MNase”).

[0260] Example 9

[0261] In FIG. 10A shows a pooled screening workflow in HEK293T cells to determine optimal segRNA targeting positions for an all-in-one protein, adapted from a previous high-density ricin biochip screen in K562 cells to identify optimal segRNAs for dCas9 - KRAB (Gilbert, Horlbeck et al., Cell 2014). First, egRNA was stably expressed in HEK293T cells by delivery of lentiviruses followed by transient transfection of a plasmid encoding the all-in-one protein (Day 0). Cells expressing the all-in-one protein were sorted (day 2) and allowed to grow further for three days. On Day 5, the cells were plated and half were collected as start-time samples and the other half were cultured for another ten days (Day 15) as end-point samples. Growth phenotype (γ) is calculated as log2 of segRNA enrichment divided by the number of cell doublings between T (initial) and T (final).

[0262] In FIG. 10B - 10E are representative plots showing growth phenotypes for four genes (ARL1, EIF6, SMC3, HEATR1) from existing dCas9 - KRAB/CRISPRi data sets in K562 cells (Gilbert, Horlbeck et al., 2014) and with the all-in protein one" (bottom graph). Each dot represents an egRNA. The CNT and annotated CpG island are shown for each gene. Functional egRNAs using an all-in-one protein cover a wider range than functional egRNAs alone, meaning a wider range of effective targeting.

[0263] Example 10

[0264] In FIG. 11A - 11B compare growth phenotypes and nucleosome positioning (as signaled by MNase) for VPS53 and VPS54, and show the location of functional egRNAs in regions lacking nucleosomes. Moreover, the range of functional egRNAs is wider when using an all-in-one protein compared to dCas9 - KRAB/CRISPRi.

[0265] Example 11

[0266] In vitro transcription of two all-in-one variants (p102 (SEQ ID NO: 14) and p112 (SEQ ID NO: 15)) demonstrates full-length synthesis of each construct (Figure 12A). In FIG. 12B is a flow cytometry graph showing the expression of p102 and p112 one day after transfection of the mRNA into HEK293T cells. In FIG. 12C shows silencing of the endogenous CLTA gene over time in HEK293T cells following transfection of mRNA expressing all-in-one protein variants p102 and p112.

[0267] Example 12

[0268] In FIG. 13A are flow cytometry plots showing the induced expression of the all-in-one protein obtained by adding doxycycline to K562 cells that stably encode the all-in-one protein under the influence of a doxycycline-inducible promoter. Protein expression was observed for four days after doxycycline induction. The dotted lines in the panels in FIG. 13A represents the baseline median fluorescence of SPB without doxycycline administration. Western blotting of the cells was performed to detect all-in-one protein expression before and after doxycycline treatment (Figure 13B). The presence of the all-in-one protein is not detected 96 hours after induction. Flow cytometry plots of CD81 and CD151 knockdown 14 days after treatment of K562 cells with doxycycline are presented in Fig. 13C - 13F. The percentage of cells with the target gene disabled is shown. There is no detectable all-in-one protein expression because there are no cells in the BFP+ quadrants. Quantification of CD81 and CD151 knockdown 14 days after doxycycline treatment or without doxycycline treatment is presented in Fig. 13G.

[0269] References

[0270] Ecco et al., Development 144, 2017. Lambert et al., Cell 172, 2018. Siddique et al., J. Mol. Biol., 425, 2013. Stepper et al., Nucleic Acids Res., 45, 2017. Shmakov et al., Nat. Rev. Microbiol. 15, 2017. CCebrian-Serrano et al., Mamm. Genome 7-8, 2017. Pulecio et al., Cell Stem Cell 21, 2017.

[0271] Unofficial sequence list

[0272]SEQ ID NO: 1 (p76 (all-in-one protein sequence version 1): KRAB (bold; from Gilbert et al., Cell, 2013, 2014), linkers (underline), dCas9 (italics), HA tag (lowercase), SV40 SNL (lowercase, italics), Dnmt3A (bold, italics; residues 612-912; from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016 ), 27 amino acid linker (underline, italics; from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold; from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase letters, bold), SFB (lowercase letters, underlining))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP GGSGGGS MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv EASGSGRASPGIPGSTR NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0273] SEQ ID NO: 2 (p90 (KRAB - dCas9 - XTEN16 - Dnmt3A - Dnmt3L - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014); linkers (underline), dCas9 (italics); tag HA (lowercase), SV40 SAL (lowercase, italics), XTEN16 (capitals, 16 amino acid sequence), Dnmt3A (bold, italics; from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), 27 amino acid linker (underline, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP GGSGGGS MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv SPG SGSETPGTSESATPES NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0274] SEQ ID NO: 3 (p91 (KRAB - dCas9 - Dnmt3A - Dnmt3L - P2A - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline), dCas9 (italics), HA tag (lowercase), SV40 SAL (lowercase, italics), Dnmt3A (bold, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), 27 linker amino acids (underline, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP GGSGGGS MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv EASGSGRASPGIPGSTR NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0275] SEQ ID NO: 4 (p92 (KRAB - dCas9 - XTEN16 - Dnmt3A - Dnmt3L - P2A - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline) , dCas9 (italics), HA tag (lowercase), SV40 SNL (lowercase, italics), XTEN16 (16 amino acid sequence), Dnmt3A (bold, italics, from Siddique et al., JMB, 2013; Stepper et al ., NAR, 2016), 27 amino acid linker (underline, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold, from Siddique et al., JMB , 2013; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP GGSGGGS MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv SPG SGSETPGTSESATPES NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0276] SEQ ID NO: 5 (p93 (KRAB - dCas9 - XTEN80 - Dnmt3A - Dnmt3L - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline), dCas9 (italics), HA tag (lowercase), SV40 SAL (lowercase, italics), XTEN80 (80 amino acid sequence), Dnmt3A (bold, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), 27 amino acid linker (underline, italics; from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold, from Siddique et al., JMB, 2013 ; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP GGSGGGS MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv SPG GGPSSGAPPPSGGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSE NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0277] SEQ ID NO: 6 (p94 (KRAB - dCas9 - XTEN80 - Dnmt3A - Dnmt3L - P2A - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline) , dCas9 (italics), HA tag (lowercase), SV40 SNL (lowercase, italics), XTEN80 (80 amino acid sequence), Dnmt3A (bold, italics, from Siddique et al., JMB, 2013; Stepper et al ., NAR, 2016), 27 amino acid linker (underline, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold, from Siddique et al., JMB , 2013; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP GGSGGGS MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv SPG GGPSSGAPPPSGGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSE NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0278] SEQ ID NO: 7 (p95 (KRAB - XTEN16 - dCas9 - Dnmt3A - Dnmt3L - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline), XTEN16 (16 amino acid sequence), dCas9 (italics), HA tag (lowercase), SV40 SAL (lowercase, italics), Dnmt3A (bold, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), 27 amino acid linker (underline, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold, from Siddique et al., JMB, 2013 ; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP SGSETPGTSESATPES MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv EASGSGRASPGIPGSTR NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0279] SEQ ID NO: 8 (p96 (KRAB - XTEN16 - dCas9 - Dnmt3A - Dnmt3L - P2A - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline) , XTEN16 (16 amino acid sequence), dCas9 (italics), HA tag (lowercase), SV40 SAL (lowercase, italics), Dnmt3A (bold, italics, from Siddique et al., JMB, 2013; Stepper et al ., NAR, 2016), 27 amino acid linker (underline, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold, from Siddique et al., JMB , 2013; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold); BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP SGSETPGTSESATPES MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv EASGSGRASPGIPGSTR NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0280] SEQ ID NO: 9 (p97 (KRAB - XTEN80 - dCas9 - Dnmt3A - Dnmt3L - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline), XTEN80 (80 amino acid sequence), dCas9 (italics); HA tag (lowercase), SV40 SAL (lowercase, italics), Dnmt3A (bold, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), 27 amino acid linker (underline, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold, from Siddique et al., JMB, 2013 ; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP GGPSSGAPPPSGGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSE MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv EASGSGRASPGIPGSTR NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0281] SEQ ID NO: 10 (p98 (KRAB - XTEN80 - dCas9 - Dnmt3A - Dnmt3L - P2A - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline) , XTEN80 (80 amino acid sequence), dCas9 (italics), HA tag (lowercase), SV40 SAL (lowercase, italics), Dnmt3A (bold, italics, from Siddique et al., JMB, 2013; Stepper et al ., NAR, 2016), 27 amino acid linker (underline, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold, from Siddique et al., JMB , 2013; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP GGPSSGAPPPSGGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSE MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv EASGSGRASPGIPGSTR NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0282] SEQ ID NO: 11 (p99 (KRAB - XTEN16 - dCas9 - XTEN80 - Dnmt3A - Dnmt3L - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), XTEN16 (sequence from 16 amino acids), dCas9 (italic), HA tag (lowercase), linkers (underline), SV40 SAL (lowercase, italic), XTEN80 (lowercase, bold, italic, 80 amino acid sequence), Dnmt3A (bold , italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), 27 amino acid linker (underlining, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016 ), Dnmt3L (underline, bold, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP SGSETPGTSESATPES MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv SPG ggpssgapppsggspagsptsteegtsesatpesgpgtstepsegsapgspagsptsteegtstepsegsapgtstepseNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0283] SEQ ID NO: 12 (p100 (KRAB - XTEN16 - dCas9 - XTEN80 - Dnmt3A - Dnmt3L - P2A - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), XTEN16 ( 16 amino acid sequence), dCas9 (italics), HA tag (lowercase), linkers (underline), SV40 SAL (lowercase, italics), XTEN80 (lowercase, bold, italics, 80 amino acid sequence), Dnmt3A ( bold, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), 27 amino acid linker (underline, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR , 2016), Dnmt3L (underline, bold, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), P2A peptide cleavage sequence (lowercase, bold), BFP (lowercase, underline) )

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP SGSETPGTSESATPES MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv SPG ggpssgapppsggspagsptsteegtsesatpesgpgtstepsegsapgspagsptsteegtstepsegsapgtstepseNHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0284] SEQ ID NO: 13 (p101 (KRAB - XTEN80 - dCas9 - XTEN16 - Dnmt3A - Dnmt3L - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline) , XTEN80 (lowercase, bold, italic, 80 amino acid sequence), dCas9 (italic), HA tag (lowercase), SV40 SAL (lowercase, italic), XTEN16 (16 amino acid sequence), Dnmt3A (bold , italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), 27 amino acid linker (underlining, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016 ), Dnmt3L (underline, bold, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP ggpssgapppsggspagsptsteegtsesatpesgpgtstepsegsapgspagsptsteegtstepsegsapgtstepse MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv SPG SGSETPGTSESATPES NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0285] SEQ ID NO: 14 (p102 (KRAB - XTEN80 - dCas9 - XTEN16 - Dnmt3A - Dnmt3L - P2A - BFP): KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline) , XTEN80 (lowercase, bold, italic, 80 amino acid sequence), dCas9 (italic), HA tag (lowercase), SV40 SAL (lowercase, italic), XTEN16 (16 amino acid sequence), Dnmt3A (bold , italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), 27 amino acid linker (underlining, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016 ), Dnmt3L (underline, bold, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), peptide cleavage sequence P2A (lowercase, bold), BFP (lowercase, underline))

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP ggpssgapppsggspagsptsteegtsesatpesgpgtstepsegsapgspagsptsteegtstepsegsapgtstepse MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SRAD ypydvpdya SGS pkkkrkv SPG SGSETPGTSESATPES NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDKLELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL SRAD pkkkrkv GSG atnfsllkqagdveenpgp GSG atnfsllkqagdveenpgp selikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianikttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghkln *

[0286] SEQ ID NO: 15 (p112 (Dnmt3A - Dnmt3L - XTEN80 - dCas9 - BFP - KRAB); KRAB (bold, from Gilbert et al., Cell, 2013, 2014), linkers (underline), XTEN80 (lowercase letters, bold, italics, 80 amino acid sequence), dCas9 (italics), HA tag (lowercase), SV40 SAL (lowercase, italics), Dnmt3A (bold, italics, from Siddique et al., JMB, 2013 ; Stepper et al., NAR, 2016), 27 amino acid linker (underline, italics, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), Dnmt3L (underline, bold, from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016), BFP (lower case, underline))

NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDK LELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV SSGNSNANSRGPSFSSGLVPLSLRGSH MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLV KNCLLLPLREYFKYFSQNSLPL ggpssgapppsggspagsptsteegtsesatpesgpgtstepsegsapgspagsptsteegtstepsegsapgtstepse MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD AypydvpdyaSLGSGS pkkkrkv ED pkkkrkv DGIGSGSNGSSGSselikenmhmklymegtvdnhhfkctsegegkpyegtqtmrikvveggplpfafdilatsflygsktfinhtqgipdffkqsfpegftwervttyedggvltatqdtslqdgcliynvkirgvnftsngpvmqkktlgweaftetlypadgglegrndmalklvggshlianik ttyrskkpaknlkmpgvyyvdyrlerikeannetyveqhevavarycdlpsklghklnGGGGGMDAKSLTAWS RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP *

[0287] SEQ ID NO: 16 (KRAB; from Gilbert et al., Cell, 2013, 2014)

DAKSLTAWSRTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQLTKPDVILRLEKGEEP

[0288] SEQ ID NO: 17 (linker)

GGSGGGS

[0289] SEQ ID NO: 18 (linker)

SGS

[0290] SEQ ID NO: 19 (linker)

EASGSGRASPGIPGSTR

[0291] SEQ ID NO: 20 (linker)

SRAD

[0292] SEQ ID NO: 21 (linker)

GSG

[0293] SEQ ID NO: 22 (linker

SPG

[0294] SEQ ID NO: 23 (dCas9)

MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYN QLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLV KLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISG VEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERM KRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQ FYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLI HQSITGLYETRIDLSQLGGD

[0295] SEQ ID NO: 24 (label HA)

YPYDVPDYA

[0296] SEQ ID NO: 25 (SV40 SYAL)

PKKKRKV

[0297] SEQ ID NO: 26 (Dnmt3A; residues 612-912; from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016)

NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDK LELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV

[0298] SEQ ID NO: 27 (27 amino acid linker; from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016)

SSGNSNANSRGPSFSSGLVPLSLRGSH

[0299] SEQ ID NO: 28 (Dnmt3L; from Siddique et al., JMB, 2013; Stepper et al., NAR, 2016)

MGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGSTQPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLV KNCLLLPLREYFKYFSQNSLPL

[0300] SEQ ID NO: 29 (P2A peptide cleavage sequence)

ATNFSLLKQAGDVEENPGP

[0301] SEQ ID NO: 30 (SFP, English “BFP”)

SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKLVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDY RLERIKEANNETYVEQHEVAVARYCDLPSKLGHKLN *

[0302] SEQ ID NO: 31 (XTEN16 (16 amino acid sequence))

SGSETPGTSESATPES

[0303] SEQ ID NO: 32 (XTEN80 (80 amino acid sequence))

GGPSSGAPPPSGGSPAGSPTSTEEGTSESATPESGPGTSTEPSEGSAPGSPAGSPTSTEEGTSTEPSEGSAPGTSTEPSE

[0304] SEQ ID NO: 33 (domain Dnmt3A - Dnmt3L)

NHDQEFDPPKVYPPVPAEKRKPIRVLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKHIQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFWLFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDK LELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFPVHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACVSSGNSNANSRGPSFSSGLVPLSLRGSHMGPMEIYKTVSAWKRQPVRVLSLFRNIDKVLKSLGFLESGSGSGGGTLKYVEDVTNVVRRDVEKWGPFDLVYGST QPLGSSCDRCPGWYMFQFHRILQYALPRQESQRPFFWIFMDNLLLTEDDQETTTRFLQTEAVTLQDVRGRDYQNAMRVWSNIPGLKSKHAPLTPKEEEYLQAQVRSRSKLDAPKVDLLVKNCLLPLREYFKYFSQNSLPL

[0305] SEQ ID NO: 34 (ddAsCfp1)

MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDHYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNAIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDKFTTYFSGFYENRKNVFSAEDISTAIPHRIVQ DNFPKFKENCHIFTRLITAVPSLREHFENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLNLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLRNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGKITKSAKE KVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLKKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKARNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKALSFEPTEKTSEGFDKMYYDYF PDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPLEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDLSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGHHGKPNLHTLYWTGLFSPENLAKTSIKLNGQ AELFYRPKSRMKRMAHRLGEKMLNKKLKDQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFHVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRSLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGY LSQVIHEIVDLMIHYQAVVVLANLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQFTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDVKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGK RIVPVIENHRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMRNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESKDLKLQNGISNQDWLAYIQELRN

[0306] SEQ ID NO: 35 (ddLbCfp1)

MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAEDYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKEIAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFSEEAKSSTSIAFRCINENLTRYISNMDIFEKVDAIF DKHEVQEIKEKILNSDYDVEDFFEGEFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVLSDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKNGPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDR RKSFKKIGSFSLEQLQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKDLLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYSKDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQ KIDKDDVNGNYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKLIDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLVEEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASL KKEELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKINTEVRVLLKHDDNPYVIGIARGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTDYHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALADLNSGFKNSRVKVEKQ VYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQNGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALDYKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQQGDIRALLCEQSDKAFYSSFMALMSLML QMRNSITGRTDVDFLISPVKNSDGIFYDSRNYEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTSVKH

[0307] SEQ ID NO: 36 (ddFnCfp1)

MYPYDVPDYASGSGMSIYQEFVNKYSLSKTLRFELIPQGKTLENIKARGLILDDEKRAKDYKKAKQIIDKYHQFFIEEILSSVCISEDLLQNYSDVYFKLKKSDDDNLQKDFKSAKDTIKKQISEYIKDSEKFKNLFNQNLIDAKKGQESDLILWLKQSKDNGIELFKANSDITDIDEA LEIIKSFKGWTTYFKGFHENRKNVYSSNDIPTSIIYRIVDDNLPKFLENKAKYESLKDKAPEAINYEQIKKDLAEELTFDIDYKTSEVNQRVFSLDEVFEIANFNNYLNQSGITKFNTIIGGKFVNGENTKRKGINEYINLYSQQINDKTLKKYKMSVLFKQILSDTESKSFVIDKLEDDSDVVTTMQSFY EQIAAFKTVEEKSIKETLSLLFDDLKAQKLDLSKIYFKNDKSLTDLSQQVFDDYSVIGTAVLEYITQQIAPKNLDNPSKKEQELIAKKTEKAKYLSLETIKLALEEFNKHRDIDKQCRFEEILANFAAIPMIFDEIAQNKDNLAQISIKYQNQGKKDLLQASAEDDVKAIKDLLDQTNNLLHKLKIFHI SQSEDKANILDKDEHFYLVFEECYFELANIVPLYNKIRNYITQKPYSDEKFKLNFENSTLANGWDKNKEPDNTAILFIKDDKYYLGVMNKKNNKIFDDKAIKENKGEGYKKIVYKLLPGANKMLPKVFFSAKSIKFYNPSEDILRIRNHSTHTKNGSPQKGYEKFEFNIEDCRKFIDFYKQSISKHPEWKDFGF RFSDTQRYNSIDEFYREVENQGYKLTFENISESYIDSVVNQGKLYLFQIYNKDFSAYSKGRPNLHTLYWKALFDERNLQDVVYKLNGEAELFYRKQSIPKKITHPAKEAIANKNKDNPKKESVFEYDLIKDKRFTEDKFFFHCPITINFKSSGANKFNDEINLLLKEKANDVHILSIARGERHLAYYTLVDG KGNIIKQDTFNIIGNDRMKTNYHDKLAAIEKDRDSARKDWKKINNIKEMKEGYLSQVVHEIAKLVIEYNAIVVFEDLNFGFKRGRFKVEKQVYQKLEKMLIEKLNYLVFKDNEFDKTGGVLRAYQLTAPFETFKKMGKQTGIIYYVPAGFTSKICPVTGFVNQLYPKYESVSKSQEFFSKFDK ICYNLDKGYFEFSFDYKNFGDKAAKGKWTIASFGSRLINFRNSDKNHNWDTREVYPTKELEKLLKDYSIEYGHGECIKAAICGESDKKFFAKLTSVLNTILQMRNSKTGTELDYLISPVADVNGNFFDSRQAPKNMPQDADANGAYHIGLKGLMLLGRIKNNQEGKKLNLVIKNEEYFEFVQNRNN

--->

LIST OF SEQUENCES

<110> The Regents of the University of California

Chen, Jin

Gilbert, Luke

Nunez, James

Weissman, Jonathan

<120> Compositions and methods of gene editing

<130> 048536-620001WO

<140> PCT/US2019/028377

<141> 2019-04-19

<150> US 62/660,023

<151> 2018-04-19

<160> 96

<170> PatentIn, version 3.5

<210> 1

<211> 2294

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 1

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Gly Gly Ser Gly Gly Gly Ser Met Asp

65 70 75 80

Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp

85 90 95

Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val

100 105 110

Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala

115 120 125

Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg

130 135 140

Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu

145 150 155 160

Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe

165 170 175

His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu

180 185 190

Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu

195 200 205

Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr

210 215 220

Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile

225 230 235 240

Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn

245 250 255

Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln

260 265 270

Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala

275 280 285

Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile

290 295 300

Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile

305 310 315 320

Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu

325 330 335

Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp

340 345 350

Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe

355 360 365

Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu

370 375 380

Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile

385 390 395 400

Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu

405 410 415

Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln

420 425 430

Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu

435 440 445

Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr

450 455 460

Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln

465 470 475 480

Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu

485 490 495

Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys

500 505 510

Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr

515 520 525

Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr

530 535 540

Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val

545 550 555 560

Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe

565 570 575

Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu

580 585 590

Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val

595 600 605

Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys

610 615 620

Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys

625 630 635 640

Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val

645 650 655

Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr

660 665 670

His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu

675 680 685

Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe

690 695 700

Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu

705 710 715 720

Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly

725 730 735

Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln

740 745 750

Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala Asn

755 760 765

Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu

770 775 780

Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu

785 790 795 800

His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu

805 810 815

Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His

820 825 830

Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr

835 840 845

Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu

850 855 860

Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu

865 870 875 880

Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn

885 890 895

Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser

900 905 910

Asp Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp

915 920 925

Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys

930 935 940

Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr

945 950 955 960

Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp

965 970 975

Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala

980 985 990

Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His

995 1000 1005

Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu

1010 1015 1020

Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser

1025 1030 1035

Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val

1040 1045 1050

Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn

1055 1060 1065

Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu

1070 1075 1080

Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys

1085 1090 1095

Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys

1100 1105 1110

Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile

1115 1120 1125

Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr

1130 1135 1140

Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe

1145 1150 1155

Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val

1160 1165 1170

Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile

1175 1180 1185

Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp

1190 1195 1200

Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala

1205 1210 1215

Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys

1220 1225 1230

Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu

1235 1240 1245

Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys

1250 1255 1260

Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys

1265 1270 1275

Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala

1280 1285 1290

Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser

1295 1300 1305

Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu

1310 1315 1320

Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu

1325 1330 1335

Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu

1340 1345 1350

Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val

1355 1360 1365

Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln

1370 1375 1380

Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala

1385 1390 1395

Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg

1400 1405 1410

Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln

1415 1420 1425

Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu

1430 1435 1440

Gly Gly Asp Ser Arg Ala Asp Tyr Pro Tyr Asp Val Pro Asp Tyr

1445 1450 1455

Ala Ser Gly Ser Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly

1460 1465 1470

Ser Gly Arg Ala Ser Pro Gly Ile Pro Gly Ser Thr Arg Asn His

1475 1480 1485

Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro Ala

1490 1495 1500

Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile

1505 1510 1515

Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp

1520 1525 1530

Arg Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly

1535 1540 1545

Met Val Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg

1550 1555 1560

Ser Val Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu

1565 1570 1575

Val Ile Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro

1580 1585 1590

Ala Arg Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu

1595 1600 1605

Phe Tyr Arg Leu Leu His Asp Ala Arg Pro Lys Glu Gly Asp Asp

1610 1615 1620

Arg Pro Phe Phe Trp Leu Phe Glu Asn Val Val Ala Met Gly Val

1625 1630 1635

Ser Asp Lys Arg Asp Ile Ser Arg Phe Leu Glu Ser Asn Pro Val

1640 1645 1650

Met Ile Asp Ala Lys Glu Val Ser Ala Ala His Arg Ala Arg Tyr

1655 1660 1665

Phe Trp Gly Asn Leu Pro Gly Met Asn Arg Pro Leu Ala Ser Thr

1670 1675 1680

Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu Glu His Gly Arg

1685 1690 1695

Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr Arg Ser Asn

1700 1705 1710

Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe Met Asn

1715 1720 1725

Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val Phe

1730 1735 1740

Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu

1745 1750 1755

Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile

1760 1765 1770

Arg His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val Ser

1775 1780 1785

Ser Gly Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser

1790 1795 1800

Gly Leu Val Pro Leu Ser Leu Arg Gly Ser His Met Gly Pro Met

1805 1810 1815

Glu Ile Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln Pro Val Arg

1820 1825 1830

Val Leu Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys Ser Leu

1835 1840 1845

Gly Phe Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr Leu Lys

1850 1855 1860

Tyr Val Glu Asp Val Thr Asn Val Val Arg Arg Asp Val Glu Lys

1865 1870 1875

Trp Gly Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro Leu Gly

1880 1885 1890

Ser Ser Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln Phe His

1895 1900 1905

Arg Ile Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln Arg Pro

1910 1915 1920

Phe Phe Trp Ile Phe Met Asp Asn Leu Leu Leu Thr Glu Asp Asp

1925 1930 1935

Gln Glu Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala Val Thr Leu

1940 1945 1950

Gln Asp Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg Val Trp

1955 1960 1965

Ser Asn Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu Thr Pro

1970 1975 1980

Lys Glu Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg Ser Lys

1985 1990 1995

Leu Asp Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys Leu Leu

2000 2005 2010

Pro Leu Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser Leu Pro

2015 2020 2025

Leu Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Gly Ser Gly

2030 2035 2040

Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu

2045 2050 2055

Asn Pro Gly Pro Ser Glu Leu Ile Lys Glu Asn Met His Met Lys

2060 2065 2070

Leu Tyr Met Glu Gly Thr Val Asp Asn His His Phe Lys Cys Thr

2075 2080 2085

Ser Glu Gly Glu Gly Lys Pro Tyr Glu Gly Thr Gln Thr Met Arg

2090 2095 2100

Ile Lys Val Val Glu Gly Gly Pro Leu Pro Phe Ala Phe Asp Ile

2105 2110 2115

Leu Ala Thr Ser Phe Leu Tyr Gly Ser Lys Thr Phe Ile Asn His

2120 2125 2130

Thr Gln Gly Ile Pro Asp Phe Phe Lys Gln Ser Phe Pro Glu Gly

2135 2140 2145

Phe Thr Trp Glu Arg Val Thr Thr Tyr Glu Asp Gly Gly Val Leu

2150 2155 2160

Thr Ala Thr Gln Asp Thr Ser Leu Gln Asp Gly Cys Leu Ile Tyr

2165 2170 2175

Asn Val Lys Ile Arg Gly Val Asn Phe Thr Ser Asn Gly Pro Val

2180 2185 2190

Met Gln Lys Lys Thr Leu Gly Trp Glu Ala Phe Thr Glu Thr Leu

2195 2200 2205

Tyr Pro Ala Asp Gly Gly Leu Glu Gly Arg Asn Asp Met Ala Leu

2210 2215 2220

Lys Leu Val Gly Gly Ser His Leu Ile Ala Asn Ile Lys Thr Thr

2225 2230 2235

Tyr Arg Ser Lys Lys Pro Ala Lys Asn Leu Lys Met Pro Gly Val

2240 2245 2250

Tyr Tyr Val Asp Tyr Arg Leu Glu Arg Ile Lys Glu Ala Asn Asn

2255 2260 2265

Glu Thr Tyr Val Glu Gln His Glu Val Ala Val Ala Arg Tyr Cys

2270 2275 2280

Asp Leu Pro Ser Lys Leu Gly His Lys Leu Asn

2285 2290

<210> 2

<211> 2296

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 2

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Gly Gly Ser Gly Gly Gly Ser Met Asp

65 70 75 80

Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp

85 90 95

Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val

100 105 110

Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala

115 120 125

Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg

130 135 140

Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu

145 150 155 160

Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe

165 170 175

His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu

180 185 190

Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu

195 200 205

Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr

210 215 220

Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile

225 230 235 240

Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn

245 250 255

Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln

260 265 270

Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala

275 280 285

Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile

290 295 300

Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile

305 310 315 320

Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu

325 330 335

Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp

340 345 350

Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe

355 360 365

Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu

370 375 380

Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile

385 390 395 400

Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu

405 410 415

Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln

420 425 430

Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu

435 440 445

Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr

450 455 460

Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln

465 470 475 480

Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu

485 490 495

Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys

500 505 510

Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr

515 520 525

Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr

530 535 540

Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val

545 550 555 560

Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe

565 570 575

Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu

580 585 590

Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val

595 600 605

Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys

610 615 620

Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys

625 630 635 640

Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val

645 650 655

Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr

660 665 670

His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu

675 680 685

Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe

690 695 700

Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu

705 710 715 720

Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly

725 730 735

Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln

740 745 750

Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala Asn

755 760 765

Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu

770 775 780

Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu

785 790 795 800

His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu

805 810 815

Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His

820 825 830

Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr

835 840 845

Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu

850 855 860

Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu

865 870 875 880

Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn

885 890 895

Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser

900 905 910

Asp Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp

915 920 925

Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys

930 935 940

Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr

945 950 955 960

Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp

965 970 975

Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala

980 985 990

Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His

995 1000 1005

Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu

1010 1015 1020

Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser

1025 1030 1035

Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val

1040 1045 1050

Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn

1055 1060 1065

Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu

1070 1075 1080

Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys

1085 1090 1095

Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys

1100 1105 1110

Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile

1115 1120 1125

Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr

1130 1135 1140

Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe

1145 1150 1155

Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val

1160 1165 1170

Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile

1175 1180 1185

Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp

1190 1195 1200

Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala

1205 1210 1215

Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys

1220 1225 1230

Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu

1235 1240 1245

Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys

1250 1255 1260

Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys

1265 1270 1275

Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala

1280 1285 1290

Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser

1295 1300 1305

Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu

1310 1315 1320

Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu

1325 1330 1335

Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu

1340 1345 1350

Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val

1355 1360 1365

Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln

1370 1375 1380

Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala

1385 1390 1395

Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg

1400 1405 1410

Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln

1415 1420 1425

Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu

1430 1435 1440

Gly Gly Asp Ser Arg Ala Asp Tyr Pro Tyr Asp Val Pro Asp Tyr

1445 1450 1455

Ala Ser Gly Ser Pro Lys Lys Lys Arg Lys Val Ser Pro Gly Ser

1460 1465 1470

Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser

1475 1480 1485

Asn His Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val

1490 1495 1500

Pro Ala Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp

1505 1510 1515

Gly Ile Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln

1520 1525 1530

Val Asp Arg Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr

1535 1540 1545

Val Gly Met Val Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp

1550 1555 1560

Val Arg Ser Val Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe

1565 1570 1575

Asp Leu Val Ile Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val

1580 1585 1590

Asn Pro Ala Arg Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe

1595 1600 1605

Phe Glu Phe Tyr Arg Leu Leu His Asp Ala Arg Pro Lys Glu Gly

1610 1615 1620

Asp Asp Arg Pro Phe Phe Trp Leu Phe Glu Asn Val Val Ala Met

1625 1630 1635

Gly Val Ser Asp Lys Arg Asp Ile Ser Arg Phe Leu Glu Ser Asn

1640 1645 1650

Pro Val Met Ile Asp Ala Lys Glu Val Ser Ala Ala His Arg Ala

1655 1660 1665

Arg Tyr Phe Trp Gly Asn Leu Pro Gly Met Asn Arg Pro Leu Ala

1670 1675 1680

Ser Thr Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu Glu His

1685 1690 1695

Gly Arg Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr Arg

1700 1705 1710

Ser Asn Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe

1715 1720 1725

Met Asn Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg

1730 1735 1740

Val Phe Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser

1745 1750 1755

Arg Leu Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro

1760 1765 1770

Val Ile Arg His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys

1775 1780 1785

Val Ser Ser Gly Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe

1790 1795 1800

Ser Ser Gly Leu Val Pro Leu Ser Leu Arg Gly Ser His Met Gly

1805 1810 1815

Pro Met Glu Ile Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln Pro

1820 1825 1830

Val Arg Val Leu Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys

1835 1840 1845

Ser Leu Gly Phe Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr

1850 1855 1860

Leu Lys Tyr Val Glu Asp Val Thr Asn Val Val Arg Arg Asp Val

1865 1870 1875

Glu Lys Trp Gly Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro

1880 1885 1890

Leu Gly Ser Ser Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln

1895 1900 1905

Phe His Arg Ile Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln

1910 1915 1920

Arg Pro Phe Phe Trp Ile Phe Met Asp Asn Leu Leu Leu Thr Glu

1925 1930 1935

Asp Asp Gln Glu Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala Val

1940 1945 1950

Thr Leu Gln Asp Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg

1955 1960 1965

Val Trp Ser Asn Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu

1970 1975 1980

Thr Pro Lys Glu Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg

1985 1990 1995

Ser Lys Leu Asp Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys

2000 2005 2010

Leu Leu Pro Leu Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser

2015 2020 2025

Leu Pro Leu Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Gly

2030 2035 2040

Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val

2045 2050 2055

Glu Glu Asn Pro Gly Pro Ser Glu Leu Ile Lys Glu Asn Met His

2060 2065 2070

Met Lys Leu Tyr Met Glu Gly Thr Val Asp Asn His His Phe Lys

2075 2080 2085

Cys Thr Ser Glu Gly Glu Gly Lys Pro Tyr Glu Gly Thr Gln Thr

2090 2095 2100

Met Arg Ile Lys Val Val Glu Gly Gly Pro Leu Pro Phe Ala Phe

2105 2110 2115

Asp Ile Leu Ala Thr Ser Phe Leu Tyr Gly Ser Lys Thr Phe Ile

2120 2125 2130

Asn His Thr Gln Gly Ile Pro Asp Phe Phe Lys Gln Ser Phe Pro

2135 2140 2145

Glu Gly Phe Thr Trp Glu Arg Val Thr Thr Tyr Glu Asp Gly Gly

2150 2155 2160

Val Leu Thr Ala Thr Gln Asp Thr Ser Leu Gln Asp Gly Cys Leu

2165 2170 2175

Ile Tyr Asn Val Lys Ile Arg Gly Val Asn Phe Thr Ser Asn Gly

2180 2185 2190

Pro Val Met Gln Lys Lys Thr Leu Gly Trp Glu Ala Phe Thr Glu

2195 2200 2205

Thr Leu Tyr Pro Ala Asp Gly Gly Leu Glu Gly Arg Asn Asp Met

2210 2215 2220

Ala Leu Lys Leu Val Gly Gly Ser His Leu Ile Ala Asn Ile Lys

2225 2230 2235

Thr Thr Tyr Arg Ser Lys Lys Pro Ala Lys Asn Leu Lys Met Pro

2240 2245 2250

Gly Val Tyr Tyr Val Asp Tyr Arg Leu Glu Arg Ile Lys Glu Ala

2255 2260 2265

Asn Asn Glu Thr Tyr Val Glu Gln His Glu Val Ala Val Ala Arg

2270 2275 2280

Tyr Cys Asp Leu Pro Ser Lys Leu Gly His Lys Leu Asn

2285 2290 2295

<210> 3

<211> 2316

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 3

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Gly Gly Ser Gly Gly Gly Ser Met Asp

65 70 75 80

Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp

85 90 95

Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val

100 105 110

Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala

115 120 125

Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg

130 135 140

Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu

145 150 155 160

Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe

165 170 175

His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu

180 185 190

Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu

195 200 205

Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr

210 215 220

Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile

225 230 235 240

Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn

245 250 255

Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln

260 265 270

Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala

275 280 285

Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile

290 295 300

Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile

305 310 315 320

Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu

325 330 335

Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp

340 345 350

Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe

355 360 365

Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu

370 375 380

Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile

385 390 395 400

Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu

405 410 415

Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln

420 425 430

Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu

435 440 445

Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr

450 455 460

Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln

465 470 475 480

Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu

485 490 495

Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys

500 505 510

Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr

515 520 525

Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr

530 535 540

Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val

545 550 555 560

Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe

565 570 575

Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu

580 585 590

Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val

595 600 605

Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys

610 615 620

Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys

625 630 635 640

Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val

645 650 655

Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr

660 665 670

His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu

675 680 685

Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe

690 695 700

Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu

705 710 715 720

Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly

725 730 735

Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln

740 745 750

Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala Asn

755 760 765

Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu

770 775 780

Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu

785 790 795 800

His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu

805 810 815

Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His

820 825 830

Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr

835 840 845

Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu

850 855 860

Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu

865 870 875 880

Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn

885 890 895

Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser

900 905 910

Asp Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp

915 920 925

Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys

930 935 940

Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr

945 950 955 960

Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp

965 970 975

Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala

980 985 990

Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His

995 1000 1005

Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu

1010 1015 1020

Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser

1025 1030 1035

Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val

1040 1045 1050

Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn

1055 1060 1065

Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu

1070 1075 1080

Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys

1085 1090 1095

Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys

1100 1105 1110

Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile

1115 1120 1125

Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr

1130 1135 1140

Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe

1145 1150 1155

Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val

1160 1165 1170

Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile

1175 1180 1185

Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp

1190 1195 1200

Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala

1205 1210 1215

Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys

1220 1225 1230

Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu

1235 1240 1245

Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys

1250 1255 1260

Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys

1265 1270 1275

Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala

1280 1285 1290

Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser

1295 1300 1305

Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu

1310 1315 1320

Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu

1325 1330 1335

Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu

1340 1345 1350

Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val

1355 1360 1365

Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln

1370 1375 1380

Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala

1385 1390 1395

Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg

1400 1405 1410

Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln

1415 1420 1425

Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu

1430 1435 1440

Gly Gly Asp Ser Arg Ala Asp Tyr Pro Tyr Asp Val Pro Asp Tyr

1445 1450 1455

Ala Ser Gly Ser Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly

1460 1465 1470

Ser Gly Arg Ala Ser Pro Gly Ile Pro Gly Ser Thr Arg Asn His

1475 1480 1485

Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro Ala

1490 1495 1500

Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile

1505 1510 1515

Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp

1520 1525 1530

Arg Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly

1535 1540 1545

Met Val Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg

1550 1555 1560

Ser Val Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu

1565 1570 1575

Val Ile Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro

1580 1585 1590

Ala Arg Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu

1595 1600 1605

Phe Tyr Arg Leu Leu His Asp Ala Arg Pro Lys Glu Gly Asp Asp

1610 1615 1620

Arg Pro Phe Phe Trp Leu Phe Glu Asn Val Val Ala Met Gly Val

1625 1630 1635

Ser Asp Lys Arg Asp Ile Ser Arg Phe Leu Glu Ser Asn Pro Val

1640 1645 1650

Met Ile Asp Ala Lys Glu Val Ser Ala Ala His Arg Ala Arg Tyr

1655 1660 1665

Phe Trp Gly Asn Leu Pro Gly Met Asn Arg Pro Leu Ala Ser Thr

1670 1675 1680

Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu Glu His Gly Arg

1685 1690 1695

Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr Arg Ser Asn

1700 1705 1710

Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe Met Asn

1715 1720 1725

Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val Phe

1730 1735 1740

Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu

1745 1750 1755

Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile

1760 1765 1770

Arg His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val Ser

1775 1780 1785

Ser Gly Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser

1790 1795 1800

Gly Leu Val Pro Leu Ser Leu Arg Gly Ser His Met Gly Pro Met

1805 1810 1815

Glu Ile Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln Pro Val Arg

1820 1825 1830

Val Leu Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys Ser Leu

1835 1840 1845

Gly Phe Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr Leu Lys

1850 1855 1860

Tyr Val Glu Asp Val Thr Asn Val Val Arg Arg Asp Val Glu Lys

1865 1870 1875

Trp Gly Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro Leu Gly

1880 1885 1890

Ser Ser Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln Phe His

1895 1900 1905

Arg Ile Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln Arg Pro

1910 1915 1920

Phe Phe Trp Ile Phe Met Asp Asn Leu Leu Leu Thr Glu Asp Asp

1925 1930 1935

Gln Glu Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala Val Thr Leu

1940 1945 1950

Gln Asp Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg Val Trp

1955 1960 1965

Ser Asn Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu Thr Pro

1970 1975 1980

Lys Glu Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg Ser Lys

1985 1990 1995

Leu Asp Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys Leu Leu

2000 2005 2010

Pro Leu Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser Leu Pro

2015 2020 2025

Leu Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Gly Ser Gly

2030 2035 2040

Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu

2045 2050 2055

Asn Pro Gly Pro Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys

2060 2065 2070

Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Ser Glu Leu Ile

2075 2080 2085

Lys Glu Asn Met His Met Lys Leu Tyr Met Glu Gly Thr Val Asp

2090 2095 2100

Asn His His Phe Lys Cys Thr Ser Glu Gly Glu Gly Lys Pro Tyr

2105 2110 2115

Glu Gly Thr Gln Thr Met Arg Ile Lys Val Val Glu Gly Gly Pro

2120 2125 2130

Leu Pro Phe Ala Phe Asp Ile Leu Ala Thr Ser Phe Leu Tyr Gly

2135 2140 2145

Ser Lys Thr Phe Ile Asn His Thr Gln Gly Ile Pro Asp Phe Phe

2150 2155 2160

Lys Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg Val Thr Thr

2165 2170 2175

Tyr Glu Asp Gly Gly Val Leu Thr Ala Thr Gln Asp Thr Ser Leu

2180 2185 2190

Gln Asp Gly Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly Val Asn

2195 2200 2205

Phe Thr Ser Asn Gly Pro Val Met Gln Lys Lys Thr Leu Gly Trp

2210 2215 2220

Glu Ala Phe Thr Glu Thr Leu Tyr Pro Ala Asp Gly Gly Leu Glu

2225 2230 2235

Gly Arg Asn Asp Met Ala Leu Lys Leu Val Gly Gly Ser His Leu

2240 2245 2250

Ile Ala Asn Ile Lys Thr Thr Tyr Arg Ser Lys Lys Pro Ala Lys

2255 2260 2265

Asn Leu Lys Met Pro Gly Val Tyr Tyr Val Asp Tyr Arg Leu Glu

2270 2275 2280

Arg Ile Lys Glu Ala Asn Asn Glu Thr Tyr Val Glu Gln His Glu

2285 2290 2295

Val Ala Val Ala Arg Tyr Cys Asp Leu Pro Ser Lys Leu Gly His

2300 2305 2310

Lys Leu Asn

2315

<210> 4

<211> 2318

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 4

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Gly Gly Ser Gly Gly Gly Ser Met Asp

65 70 75 80

Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp

85 90 95

Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val

100 105 110

Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala

115 120 125

Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg

130 135 140

Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu

145 150 155 160

Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe

165 170 175

His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu

180 185 190

Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu

195 200 205

Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr

210 215 220

Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile

225 230 235 240

Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn

245 250 255

Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln

260 265 270

Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala

275 280 285

Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile

290 295 300

Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile

305 310 315 320

Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu

325 330 335

Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp

340 345 350

Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe

355 360 365

Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu

370 375 380

Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile

385 390 395 400

Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu

405 410 415

Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln

420 425 430

Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu

435 440 445

Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr

450 455 460

Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln

465 470 475 480

Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu

485 490 495

Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys

500 505 510

Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr

515 520 525

Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr

530 535 540

Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val

545 550 555 560

Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe

565 570 575

Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu

580 585 590

Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val

595 600 605

Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys

610 615 620

Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys

625 630 635 640

Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val

645 650 655

Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr

660 665 670

His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu

675 680 685

Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe

690 695 700

Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu

705 710 715 720

Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly

725 730 735

Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln

740 745 750

Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala Asn

755 760 765

Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu

770 775 780

Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu

785 790 795 800

His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu

805 810 815

Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His

820 825 830

Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr

835 840 845

Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu

850 855 860

Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu

865 870 875 880

Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn

885 890 895

Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser

900 905 910

Asp Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp

915 920 925

Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys

930 935 940

Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr

945 950 955 960

Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp

965 970 975

Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala

980 985 990

Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His

995 1000 1005

Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu

1010 1015 1020

Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser

1025 1030 1035

Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val

1040 1045 1050

Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn

1055 1060 1065

Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu

1070 1075 1080

Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys

1085 1090 1095

Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys

1100 1105 1110

Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile

1115 1120 1125

Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr

1130 1135 1140

Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe

1145 1150 1155

Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val

1160 1165 1170

Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile

1175 1180 1185

Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp

1190 1195 1200

Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala

1205 1210 1215

Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys

1220 1225 1230

Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu

1235 1240 1245

Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys

1250 1255 1260

Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys

1265 1270 1275

Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala

1280 1285 1290

Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser

1295 1300 1305

Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu

1310 1315 1320

Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu

1325 1330 1335

Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu

1340 1345 1350

Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val

1355 1360 1365

Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln

1370 1375 1380

Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala

1385 1390 1395

Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg

1400 1405 1410

Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln

1415 1420 1425

Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu

1430 1435 1440

Gly Gly Asp Ser Arg Ala Asp Tyr Pro Tyr Asp Val Pro Asp Tyr

1445 1450 1455

Ala Ser Gly Ser Pro Lys Lys Lys Arg Lys Val Ser Pro Gly Ser

1460 1465 1470

Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser

1475 1480 1485

Asn His Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val

1490 1495 1500

Pro Ala Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp

1505 1510 1515

Gly Ile Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln

1520 1525 1530

Val Asp Arg Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr

1535 1540 1545

Val Gly Met Val Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp

1550 1555 1560

Val Arg Ser Val Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe

1565 1570 1575

Asp Leu Val Ile Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val

1580 1585 1590

Asn Pro Ala Arg Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe

1595 1600 1605

Phe Glu Phe Tyr Arg Leu Leu His Asp Ala Arg Pro Lys Glu Gly

1610 1615 1620

Asp Asp Arg Pro Phe Phe Trp Leu Phe Glu Asn Val Val Ala Met

1625 1630 1635

Gly Val Ser Asp Lys Arg Asp Ile Ser Arg Phe Leu Glu Ser Asn

1640 1645 1650

Pro Val Met Ile Asp Ala Lys Glu Val Ser Ala Ala His Arg Ala

1655 1660 1665

Arg Tyr Phe Trp Gly Asn Leu Pro Gly Met Asn Arg Pro Leu Ala

1670 1675 1680

Ser Thr Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu Glu His

1685 1690 1695

Gly Arg Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr Arg

1700 1705 1710

Ser Asn Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe

1715 1720 1725

Met Asn Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg

1730 1735 1740

Val Phe Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser

1745 1750 1755

Arg Leu Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro

1760 1765 1770

Val Ile Arg His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys

1775 1780 1785

Val Ser Ser Gly Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe

1790 1795 1800

Ser Ser Gly Leu Val Pro Leu Ser Leu Arg Gly Ser His Met Gly

1805 1810 1815

Pro Met Glu Ile Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln Pro

1820 1825 1830

Val Arg Val Leu Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys

1835 1840 1845

Ser Leu Gly Phe Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr

1850 1855 1860

Leu Lys Tyr Val Glu Asp Val Thr Asn Val Val Arg Arg Asp Val

1865 1870 1875

Glu Lys Trp Gly Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro

1880 1885 1890

Leu Gly Ser Ser Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln

1895 1900 1905

Phe His Arg Ile Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln

1910 1915 1920

Arg Pro Phe Phe Trp Ile Phe Met Asp Asn Leu Leu Leu Thr Glu

1925 1930 1935

Asp Asp Gln Glu Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala Val

1940 1945 1950

Thr Leu Gln Asp Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg

1955 1960 1965

Val Trp Ser Asn Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu

1970 1975 1980

Thr Pro Lys Glu Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg

1985 1990 1995

Ser Lys Leu Asp Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys

2000 2005 2010

Leu Leu Pro Leu Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser

2015 2020 2025

Leu Pro Leu Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Gly

2030 2035 2040

Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val

2045 2050 2055

Glu Glu Asn Pro Gly Pro Gly Ser Gly Ala Thr Asn Phe Ser Leu

2060 2065 2070

Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Ser Glu

2075 2080 2085

Leu Ile Lys Glu Asn Met His Met Lys Leu Tyr Met Glu Gly Thr

2090 2095 2100

Val Asp Asn His His Phe Lys Cys Thr Ser Glu Gly Glu Gly Lys

2105 2110 2115

Pro Tyr Glu Gly Thr Gln Thr Met Arg Ile Lys Val Val Glu Gly

2120 2125 2130

Gly Pro Leu Pro Phe Ala Phe Asp Ile Leu Ala Thr Ser Phe Leu

2135 2140 2145

Tyr Gly Ser Lys Thr Phe Ile Asn His Thr Gln Gly Ile Pro Asp

2150 2155 2160

Phe Phe Lys Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg Val

2165 2170 2175

Thr Thr Tyr Glu Asp Gly Gly Val Leu Thr Ala Thr Gln Asp Thr

2180 2185 2190

Ser Leu Gln Asp Gly Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly

2195 2200 2205

Val Asn Phe Thr Ser Asn Gly Pro Val Met Gln Lys Lys Thr Leu

2210 2215 2220

Gly Trp Glu Ala Phe Thr Glu Thr Leu Tyr Pro Ala Asp Gly Gly

2225 2230 2235

Leu Glu Gly Arg Asn Asp Met Ala Leu Lys Leu Val Gly Gly Ser

2240 2245 2250

His Leu Ile Ala Asn Ile Lys Thr Thr Tyr Arg Ser Lys Lys Pro

2255 2260 2265

Ala Lys Asn Leu Lys Met Pro Gly Val Tyr Tyr Val Asp Tyr Arg

2270 2275 2280

Leu Glu Arg Ile Lys Glu Ala Asn Asn Glu Thr Tyr Val Glu Gln

2285 2290 2295

His Glu Val Ala Val Ala Arg Tyr Cys Asp Leu Pro Ser Lys Leu

2300 2305 2310

Gly His Lys Leu Asn

2315

<210> 5

<211> 2360

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 5

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Gly Gly Ser Gly Gly Gly Ser Met Asp

65 70 75 80

Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp

85 90 95

Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val

100 105 110

Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala

115 120 125

Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg

130 135 140

Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu

145 150 155 160

Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe

165 170 175

His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu

180 185 190

Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu

195 200 205

Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr

210 215 220

Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile

225 230 235 240

Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn

245 250 255

Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln

260 265 270

Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala

275 280 285

Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile

290 295 300

Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile

305 310 315 320

Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu

325 330 335

Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp

340 345 350

Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe

355 360 365

Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu

370 375 380

Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile

385 390 395 400

Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu

405 410 415

Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln

420 425 430

Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu

435 440 445

Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr

450 455 460

Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln

465 470 475 480

Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu

485 490 495

Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys

500 505 510

Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr

515 520 525

Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr

530 535 540

Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val

545 550 555 560

Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe

565 570 575

Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu

580 585 590

Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val

595 600 605

Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys

610 615 620

Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys

625 630 635 640

Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val

645 650 655

Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr

660 665 670

His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu

675 680 685

Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe

690 695 700

Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu

705 710 715 720

Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly

725 730 735

Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln

740 745 750

Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala Asn

755 760 765

Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu

770 775 780

Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu

785 790 795 800

His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu

805 810 815

Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His

820 825 830

Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr

835 840 845

Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu

850 855 860

Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu

865 870 875 880

Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn

885 890 895

Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser

900 905 910

Asp Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp

915 920 925

Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys

930 935 940

Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr

945 950 955 960

Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp

965 970 975

Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala

980 985 990

Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His

995 1000 1005

Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu

1010 1015 1020

Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser

1025 1030 1035

Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val

1040 1045 1050

Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn

1055 1060 1065

Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu

1070 1075 1080

Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys

1085 1090 1095

Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys

1100 1105 1110

Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile

1115 1120 1125

Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr

1130 1135 1140

Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe

1145 1150 1155

Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val

1160 1165 1170

Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile

1175 1180 1185

Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp

1190 1195 1200

Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala

1205 1210 1215

Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys

1220 1225 1230

Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu

1235 1240 1245

Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys

1250 1255 1260

Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys

1265 1270 1275

Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala

1280 1285 1290

Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser

1295 1300 1305

Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu

1310 1315 1320

Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu

1325 1330 1335

Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu

1340 1345 1350

Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val

1355 1360 1365

Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln

1370 1375 1380

Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala

1385 1390 1395

Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg

1400 1405 1410

Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln

1415 1420 1425

Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu

1430 1435 1440

Gly Gly Asp Ser Arg Ala Asp Tyr Pro Tyr Asp Val Pro Asp Tyr

1445 1450 1455

Ala Ser Gly Ser Pro Lys Lys Lys Arg Lys Val Ser Pro Gly Gly

1460 1465 1470

Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser Gly Gly Ser Pro Ala

1475 1480 1485

Gly Ser Pro Thr Ser Thr Glu Glu Gly Thr Ser Glu Ser Ala Thr

1490 1495 1500

Pro Glu Ser Gly Pro Gly Thr Ser Thr Glu Pro Ser Glu Gly Ser

1505 1510 1515

Ala Pro Gly Ser Pro Ala Gly Ser Pro Thr Ser Thr Glu Glu Gly

1520 1525 1530

Thr Ser Thr Glu Pro Ser Glu Gly Ser Ala Pro Gly Thr Ser Thr

1535 1540 1545

Glu Pro Ser Glu Asn His Asp Gln Glu Phe Asp Pro Pro Lys Val

1550 1555 1560

Tyr Pro Pro Val Pro Ala Glu Lys Arg Lys Pro Ile Arg Val Leu

1565 1570 1575

Ser Leu Phe Asp Gly Ile Ala Thr Gly Leu Leu Val Leu Lys Asp

1580 1585 1590

Leu Gly Ile Gln Val Asp Arg Tyr Ile Ala Ser Glu Val Cys Glu

1595 1600 1605

Asp Ser Ile Thr Val Gly Met Val Arg His Gln Gly Lys Ile Met

1610 1615 1620

Tyr Val Gly Asp Val Arg Ser Val Thr Gln Lys His Ile Gln Glu

1625 1630 1635

Trp Gly Pro Phe Asp Leu Val Ile Gly Gly Ser Pro Cys Asn Asp

1640 1645 1650

Leu Ser Ile Val Asn Pro Ala Arg Lys Gly Leu Tyr Glu Gly Thr

1655 1660 1665

Gly Arg Leu Phe Phe Glu Phe Tyr Arg Leu Leu His Asp Ala Arg

1670 1675 1680

Pro Lys Glu Gly Asp Asp Arg Pro Phe Phe Trp Leu Phe Glu Asn

1685 1690 1695

Val Val Ala Met Gly Val Ser Asp Lys Arg Asp Ile Ser Arg Phe

1700 1705 1710

Leu Glu Ser Asn Pro Val Met Ile Asp Ala Lys Glu Val Ser Ala

1715 1720 1725

Ala His Arg Ala Arg Tyr Phe Trp Gly Asn Leu Pro Gly Met Asn

1730 1735 1740

Arg Pro Leu Ala Ser Thr Val Asn Asp Lys Leu Glu Leu Gln Glu

1745 1750 1755

Cys Leu Glu His Gly Arg Ile Ala Lys Phe Ser Lys Val Arg Thr

1760 1765 1770

Ile Thr Thr Arg Ser Asn Ser Ile Lys Gln Gly Lys Asp Gln His

1775 1780 1785

Phe Pro Val Phe Met Asn Glu Lys Glu Asp Ile Leu Trp Cys Thr

1790 1795 1800

Glu Met Glu Arg Val Phe Gly Phe Pro Val His Tyr Thr Asp Val

1805 1810 1815

Ser Asn Met Ser Arg Leu Ala Arg Gln Arg Leu Leu Gly Arg Ser

1820 1825 1830

Trp Ser Val Pro Val Ile Arg His Leu Phe Ala Pro Leu Lys Glu

1835 1840 1845

Tyr Phe Ala Cys Val Ser Ser Gly Asn Ser Asn Ala Asn Ser Arg

1850 1855 1860

Gly Pro Ser Phe Ser Ser Gly Leu Val Pro Leu Ser Leu Arg Gly

1865 1870 1875

Ser His Met Gly Pro Met Glu Ile Tyr Lys Thr Val Ser Ala Trp

1880 1885 1890

Lys Arg Gln Pro Val Arg Val Leu Ser Leu Phe Arg Asn Ile Asp

1895 1900 1905

Lys Val Leu Lys Ser Leu Gly Phe Leu Glu Ser Gly Ser Gly Ser

1910 1915 1920

Gly Gly Gly Thr Leu Lys Tyr Val Glu Asp Val Thr Asn Val Val

1925 1930 1935

Arg Arg Asp Val Glu Lys Trp Gly Pro Phe Asp Leu Val Tyr Gly

1940 1945 1950

Ser Thr Gln Pro Leu Gly Ser Ser Cys Asp Arg Cys Pro Gly Trp

1955 1960 1965

Tyr Met Phe Gln Phe His Arg Ile Leu Gln Tyr Ala Leu Pro Arg

1970 1975 1980

Gln Glu Ser Gln Arg Pro Phe Phe Trp Ile Phe Met Asp Asn Leu

1985 1990 1995

Leu Leu Thr Glu Asp Asp Gln Glu Thr Thr Thr Arg Phe Leu Gln

2000 2005 2010

Thr Glu Ala Val Thr Leu Gln Asp Val Arg Gly Arg Asp Tyr Gln

2015 2020 2025

Asn Ala Met Arg Val Trp Ser Asn Ile Pro Gly Leu Lys Ser Lys

2030 2035 2040

His Ala Pro Leu Thr Pro Lys Glu Glu Glu Tyr Leu Gln Ala Gln

2045 2050 2055

Val Arg Ser Arg Ser Lys Leu Asp Ala Pro Lys Val Asp Leu Leu

2060 2065 2070

Val Lys Asn Cys Leu Leu Pro Leu Arg Glu Tyr Phe Lys Tyr Phe

2075 2080 2085

Ser Gln Asn Ser Leu Pro Leu Ser Arg Ala Asp Pro Lys Lys Lys

2090 2095 2100

Arg Lys Val Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln

2105 2110 2115

Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Ser Glu Leu Ile Lys

2120 2125 2130

Glu Asn Met His Met Lys Leu Tyr Met Glu Gly Thr Val Asp Asn

2135 2140 2145

His His Phe Lys Cys Thr Ser Glu Gly Glu Gly Lys Pro Tyr Glu

2150 2155 2160

Gly Thr Gln Thr Met Arg Ile Lys Val Val Glu Gly Gly Pro Leu

2165 2170 2175

Pro Phe Ala Phe Asp Ile Leu Ala Thr Ser Phe Leu Tyr Gly Ser

2180 2185 2190

Lys Thr Phe Ile Asn His Thr Gln Gly Ile Pro Asp Phe Phe Lys

2195 2200 2205

Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg Val Thr Thr Tyr

2210 2215 2220

Glu Asp Gly Gly Val Leu Thr Ala Thr Gln Asp Thr Ser Leu Gln

2225 2230 2235

Asp Gly Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly Val Asn Phe

2240 2245 2250

Thr Ser Asn Gly Pro Val Met Gln Lys Lys Thr Leu Gly Trp Glu

2255 2260 2265

Ala Phe Thr Glu Thr Leu Tyr Pro Ala Asp Gly Gly Leu Glu Gly

2270 2275 2280

Arg Asn Asp Met Ala Leu Lys Leu Val Gly Gly Ser His Leu Ile

2285 2290 2295

Ala Asn Ile Lys Thr Thr Tyr Arg Ser Lys Lys Pro Ala Lys Asn

2300 2305 2310

Leu Lys Met Pro Gly Val Tyr Tyr Val Asp Tyr Arg Leu Glu Arg

2315 2320 2325

Ile Lys Glu Ala Asn Asn Glu Thr Tyr Val Glu Gln His Glu Val

2330 2335 2340

Ala Val Ala Arg Tyr Cys Asp Leu Pro Ser Lys Leu Gly His Lys

2345 2350 2355

Leu Asn

2360

<210> 6

<211> 2382

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 6

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Gly Gly Ser Gly Gly Gly Ser Met Asp

65 70 75 80

Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp

85 90 95

Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val

100 105 110

Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala

115 120 125

Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg

130 135 140

Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu

145 150 155 160

Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe

165 170 175

His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu

180 185 190

Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu

195 200 205

Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr

210 215 220

Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile

225 230 235 240

Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn

245 250 255

Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln

260 265 270

Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala

275 280 285

Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile

290 295 300

Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile

305 310 315 320

Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu

325 330 335

Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp

340 345 350

Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe

355 360 365

Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu

370 375 380

Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile

385 390 395 400

Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu

405 410 415

Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln

420 425 430

Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu

435 440 445

Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr

450 455 460

Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln

465 470 475 480

Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu

485 490 495

Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys

500 505 510

Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr

515 520 525

Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr

530 535 540

Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val

545 550 555 560

Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe

565 570 575

Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu

580 585 590

Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val

595 600 605

Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys

610 615 620

Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys

625 630 635 640

Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val

645 650 655

Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr

660 665 670

His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu

675 680 685

Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe

690 695 700

Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu

705 710 715 720

Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly

725 730 735

Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln

740 745 750

Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala Asn

755 760 765

Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu

770 775 780

Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu

785 790 795 800

His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu

805 810 815

Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His

820 825 830

Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr

835 840 845

Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu

850 855 860

Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu

865 870 875 880

Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn

885 890 895

Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser

900 905 910

Asp Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp

915 920 925

Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys

930 935 940

Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr

945 950 955 960

Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp

965 970 975

Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala

980 985 990

Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His

995 1000 1005

Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu

1010 1015 1020

Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser

1025 1030 1035

Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val

1040 1045 1050

Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn

1055 1060 1065

Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu

1070 1075 1080

Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys

1085 1090 1095

Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys

1100 1105 1110

Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile

1115 1120 1125

Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr

1130 1135 1140

Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe

1145 1150 1155

Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val

1160 1165 1170

Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile

1175 1180 1185

Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp

1190 1195 1200

Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala

1205 1210 1215

Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys

1220 1225 1230

Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu

1235 1240 1245

Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys

1250 1255 1260

Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys

1265 1270 1275

Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala

1280 1285 1290

Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser

1295 1300 1305

Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu

1310 1315 1320

Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu

1325 1330 1335

Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu

1340 1345 1350

Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val

1355 1360 1365

Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln

1370 1375 1380

Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala

1385 1390 1395

Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg

1400 1405 1410

Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln

1415 1420 1425

Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu

1430 1435 1440

Gly Gly Asp Ser Arg Ala Asp Tyr Pro Tyr Asp Val Pro Asp Tyr

1445 1450 1455

Ala Ser Gly Ser Pro Lys Lys Lys Arg Lys Val Ser Pro Gly Gly

1460 1465 1470

Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser Gly Gly Ser Pro Ala

1475 1480 1485

Gly Ser Pro Thr Ser Thr Glu Glu Gly Thr Ser Glu Ser Ala Thr

1490 1495 1500

Pro Glu Ser Gly Pro Gly Thr Ser Thr Glu Pro Ser Glu Gly Ser

1505 1510 1515

Ala Pro Gly Ser Pro Ala Gly Ser Pro Thr Ser Thr Glu Glu Gly

1520 1525 1530

Thr Ser Thr Glu Pro Ser Glu Gly Ser Ala Pro Gly Thr Ser Thr

1535 1540 1545

Glu Pro Ser Glu Asn His Asp Gln Glu Phe Asp Pro Pro Lys Val

1550 1555 1560

Tyr Pro Pro Val Pro Ala Glu Lys Arg Lys Pro Ile Arg Val Leu

1565 1570 1575

Ser Leu Phe Asp Gly Ile Ala Thr Gly Leu Leu Val Leu Lys Asp

1580 1585 1590

Leu Gly Ile Gln Val Asp Arg Tyr Ile Ala Ser Glu Val Cys Glu

1595 1600 1605

Asp Ser Ile Thr Val Gly Met Val Arg His Gln Gly Lys Ile Met

1610 1615 1620

Tyr Val Gly Asp Val Arg Ser Val Thr Gln Lys His Ile Gln Glu

1625 1630 1635

Trp Gly Pro Phe Asp Leu Val Ile Gly Gly Ser Pro Cys Asn Asp

1640 1645 1650

Leu Ser Ile Val Asn Pro Ala Arg Lys Gly Leu Tyr Glu Gly Thr

1655 1660 1665

Gly Arg Leu Phe Phe Glu Phe Tyr Arg Leu Leu His Asp Ala Arg

1670 1675 1680

Pro Lys Glu Gly Asp Asp Arg Pro Phe Phe Trp Leu Phe Glu Asn

1685 1690 1695

Val Val Ala Met Gly Val Ser Asp Lys Arg Asp Ile Ser Arg Phe

1700 1705 1710

Leu Glu Ser Asn Pro Val Met Ile Asp Ala Lys Glu Val Ser Ala

1715 1720 1725

Ala His Arg Ala Arg Tyr Phe Trp Gly Asn Leu Pro Gly Met Asn

1730 1735 1740

Arg Pro Leu Ala Ser Thr Val Asn Asp Lys Leu Glu Leu Gln Glu

1745 1750 1755

Cys Leu Glu His Gly Arg Ile Ala Lys Phe Ser Lys Val Arg Thr

1760 1765 1770

Ile Thr Thr Arg Ser Asn Ser Ile Lys Gln Gly Lys Asp Gln His

1775 1780 1785

Phe Pro Val Phe Met Asn Glu Lys Glu Asp Ile Leu Trp Cys Thr

1790 1795 1800

Glu Met Glu Arg Val Phe Gly Phe Pro Val His Tyr Thr Asp Val

1805 1810 1815

Ser Asn Met Ser Arg Leu Ala Arg Gln Arg Leu Leu Gly Arg Ser

1820 1825 1830

Trp Ser Val Pro Val Ile Arg His Leu Phe Ala Pro Leu Lys Glu

1835 1840 1845

Tyr Phe Ala Cys Val Ser Ser Gly Asn Ser Asn Ala Asn Ser Arg

1850 1855 1860

Gly Pro Ser Phe Ser Ser Gly Leu Val Pro Leu Ser Leu Arg Gly

1865 1870 1875

Ser His Met Gly Pro Met Glu Ile Tyr Lys Thr Val Ser Ala Trp

1880 1885 1890

Lys Arg Gln Pro Val Arg Val Leu Ser Leu Phe Arg Asn Ile Asp

1895 1900 1905

Lys Val Leu Lys Ser Leu Gly Phe Leu Glu Ser Gly Ser Gly Ser

1910 1915 1920

Gly Gly Gly Thr Leu Lys Tyr Val Glu Asp Val Thr Asn Val Val

1925 1930 1935

Arg Arg Asp Val Glu Lys Trp Gly Pro Phe Asp Leu Val Tyr Gly

1940 1945 1950

Ser Thr Gln Pro Leu Gly Ser Ser Cys Asp Arg Cys Pro Gly Trp

1955 1960 1965

Tyr Met Phe Gln Phe His Arg Ile Leu Gln Tyr Ala Leu Pro Arg

1970 1975 1980

Gln Glu Ser Gln Arg Pro Phe Phe Trp Ile Phe Met Asp Asn Leu

1985 1990 1995

Leu Leu Thr Glu Asp Asp Gln Glu Thr Thr Thr Arg Phe Leu Gln

2000 2005 2010

Thr Glu Ala Val Thr Leu Gln Asp Val Arg Gly Arg Asp Tyr Gln

2015 2020 2025

Asn Ala Met Arg Val Trp Ser Asn Ile Pro Gly Leu Lys Ser Lys

2030 2035 2040

His Ala Pro Leu Thr Pro Lys Glu Glu Glu Tyr Leu Gln Ala Gln

2045 2050 2055

Val Arg Ser Arg Ser Lys Leu Asp Ala Pro Lys Val Asp Leu Leu

2060 2065 2070

Val Lys Asn Cys Leu Leu Pro Leu Arg Glu Tyr Phe Lys Tyr Phe

2075 2080 2085

Ser Gln Asn Ser Leu Pro Leu Ser Arg Ala Asp Pro Lys Lys Lys

2090 2095 2100

Arg Lys Val Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln

2105 2110 2115

Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Gly Ser Gly Ala Thr

2120 2125 2130

Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro

2135 2140 2145

Gly Pro Ser Glu Leu Ile Lys Glu Asn Met His Met Lys Leu Tyr

2150 2155 2160

Met Glu Gly Thr Val Asp Asn His His Phe Lys Cys Thr Ser Glu

2165 2170 2175

Gly Glu Gly Lys Pro Tyr Glu Gly Thr Gln Thr Met Arg Ile Lys

2180 2185 2190

Val Val Glu Gly Gly Pro Leu Pro Phe Ala Phe Asp Ile Leu Ala

2195 2200 2205

Thr Ser Phe Leu Tyr Gly Ser Lys Thr Phe Ile Asn His Thr Gln

2210 2215 2220

Gly Ile Pro Asp Phe Phe Lys Gln Ser Phe Pro Glu Gly Phe Thr

2225 2230 2235

Trp Glu Arg Val Thr Thr Tyr Glu Asp Gly Gly Val Leu Thr Ala

2240 2245 2250

Thr Gln Asp Thr Ser Leu Gln Asp Gly Cys Leu Ile Tyr Asn Val

2255 2260 2265

Lys Ile Arg Gly Val Asn Phe Thr Ser Asn Gly Pro Val Met Gln

2270 2275 2280

Lys Lys Thr Leu Gly Trp Glu Ala Phe Thr Glu Thr Leu Tyr Pro

2285 2290 2295

Ala Asp Gly Gly Leu Glu Gly Arg Asn Asp Met Ala Leu Lys Leu

2300 2305 2310

Val Gly Gly Ser His Leu Ile Ala Asn Ile Lys Thr Thr Tyr Arg

2315 2320 2325

Ser Lys Lys Pro Ala Lys Asn Leu Lys Met Pro Gly Val Tyr Tyr

2330 2335 2340

Val Asp Tyr Arg Leu Glu Arg Ile Lys Glu Ala Asn Asn Glu Thr

2345 2350 2355

Tyr Val Glu Gln His Glu Val Ala Val Ala Arg Tyr Cys Asp Leu

2360 2365 2370

Pro Ser Lys Leu Gly His Lys Leu Asn

2375 2380

<210> 7

<211> 2303

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 7

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Ser Gly Ser Glu Thr Pro Gly Thr Ser

65 70 75 80

Glu Ser Ala Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu

85 90 95

Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr

100 105 110

Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His

115 120 125

Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu

130 135 140

Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr

145 150 155 160

Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu

165 170 175

Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe

180 185 190

Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn

195 200 205

Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His

210 215 220

Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu

225 230 235 240

Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu

245 250 255

Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe

260 265 270

Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile

275 280 285

Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser

290 295 300

Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys

305 310 315 320

Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr

325 330 335

Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln

340 345 350

Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln

355 360 365

Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser

370 375 380

Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr

385 390 395 400

Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His

405 410 415

Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu

420 425 430

Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly

435 440 445

Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys

450 455 460

Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu

465 470 475 480

Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser

485 490 495

Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg

500 505 510

Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu

515 520 525

Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg

530 535 540

Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile

545 550 555 560

Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln

565 570 575

Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu

580 585 590

Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr

595 600 605

Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro

610 615 620

Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe

625 630 635 640

Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe

645 650 655

Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp

660 665 670

Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile

675 680 685

Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu

690 695 700

Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu

705 710 715 720

Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys

725 730 735

Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys

740 745 750

Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp

755 760 765

Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile

770 775 780

His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val

785 790 795 800

Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly

805 810 815

Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp

820 825 830

Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile

835 840 845

Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser

850 855 860

Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser

865 870 875 880

Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu

885 890 895

Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp

900 905 910

Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp Ala Ile

915 920 925

Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu

930 935 940

Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu

945 950 955 960

Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala

965 970 975

Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg

980 985 990

Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu

995 1000 1005

Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp

1010 1015 1020

Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg

1025 1030 1035

Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe

1040 1045 1050

Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr

1055 1060 1065

His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala

1070 1075 1080

Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly

1085 1090 1095

Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu

1100 1105 1110

Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn

1115 1120 1125

Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu

1130 1135 1140

Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu

1145 1150 1155

Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val

1160 1165 1170

Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln

1175 1180 1185

Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser

1190 1195 1200

Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr

1205 1210 1215

Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val

1220 1225 1230

Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys

1235 1240 1245

Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys

1250 1255 1260

Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys

1265 1270 1275

Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu Leu

1280 1285 1290

Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu Leu Gln

1295 1300 1305

Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn Phe Leu

1310 1315 1320

Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro Glu Asp

1325 1330 1335

Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His Tyr Leu

1340 1345 1350

Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg Val Ile

1355 1360 1365

Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys

1370 1375 1380

His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile Ile His

1385 1390 1395

Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys Tyr

1400 1405 1410

Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu

1415 1420 1425

Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr

1430 1435 1440

Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp Ser Arg Ala

1445 1450 1455

Asp Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Gly Ser Pro Lys

1460 1465 1470

Lys Lys Arg Lys Val Glu Ala Ser Gly Ser Gly Arg Ala Ser Pro

1475 1480 1485

Gly Ile Pro Gly Ser Thr Arg Asn His Asp Gln Glu Phe Asp Pro

1490 1495 1500

Pro Lys Val Tyr Pro Pro Val Pro Ala Glu Lys Arg Lys Pro Ile

1505 1510 1515

Arg Val Leu Ser Leu Phe Asp Gly Ile Ala Thr Gly Leu Leu Val

1520 1525 1530

Leu Lys Asp Leu Gly Ile Gln Val Asp Arg Tyr Ile Ala Ser Glu

1535 1540 1545

Val Cys Glu Asp Ser Ile Thr Val Gly Met Val Arg His Gln Gly

1550 1555 1560

Lys Ile Met Tyr Val Gly Asp Val Arg Ser Val Thr Gln Lys His

1565 1570 1575

Ile Gln Glu Trp Gly Pro Phe Asp Leu Val Ile Gly Gly Ser Pro

1580 1585 1590

Cys Asn Asp Leu Ser Ile Val Asn Pro Ala Arg Lys Gly Leu Tyr

1595 1600 1605

Glu Gly Thr Gly Arg Leu Phe Phe Glu Phe Tyr Arg Leu Leu His

1610 1615 1620

Asp Ala Arg Pro Lys Glu Gly Asp Asp Arg Pro Phe Phe Trp Leu

1625 1630 1635

Phe Glu Asn Val Val Ala Met Gly Val Ser Asp Lys Arg Asp Ile

1640 1645 1650

Ser Arg Phe Leu Glu Ser Asn Pro Val Met Ile Asp Ala Lys Glu

1655 1660 1665

Val Ser Ala Ala His Arg Ala Arg Tyr Phe Trp Gly Asn Leu Pro

1670 1675 1680

Gly Met Asn Arg Pro Leu Ala Ser Thr Val Asn Asp Lys Leu Glu

1685 1690 1695

Leu Gln Glu Cys Leu Glu His Gly Arg Ile Ala Lys Phe Ser Lys

1700 1705 1710

Val Arg Thr Ile Thr Thr Arg Ser Asn Ser Ile Lys Gln Gly Lys

1715 1720 1725

Asp Gln His Phe Pro Val Phe Met Asn Glu Lys Glu Asp Ile Leu

1730 1735 1740

Trp Cys Thr Glu Met Glu Arg Val Phe Gly Phe Pro Val His Tyr

1745 1750 1755

Thr Asp Val Ser Asn Met Ser Arg Leu Ala Arg Gln Arg Leu Leu

1760 1765 1770

Gly Arg Ser Trp Ser Val Pro Val Ile Arg His Leu Phe Ala Pro

1775 1780 1785

Leu Lys Glu Tyr Phe Ala Cys Val Ser Ser Gly Asn Ser Asn Ala

1790 1795 1800

Asn Ser Arg Gly Pro Ser Phe Ser Ser Gly Leu Val Pro Leu Ser

1805 1810 1815

Leu Arg Gly Ser His Met Gly Pro Met Glu Ile Tyr Lys Thr Val

1820 1825 1830

Ser Ala Trp Lys Arg Gln Pro Val Arg Val Leu Ser Leu Phe Arg

1835 1840 1845

Asn Ile Asp Lys Val Leu Lys Ser Leu Gly Phe Leu Glu Ser Gly

1850 1855 1860

Ser Gly Ser Gly Gly Gly Thr Leu Lys Tyr Val Glu Asp Val Thr

1865 1870 1875

Asn Val Val Arg Arg Asp Val Glu Lys Trp Gly Pro Phe Asp Leu

1880 1885 1890

Val Tyr Gly Ser Thr Gln Pro Leu Gly Ser Ser Cys Asp Arg Cys

1895 1900 1905

Pro Gly Trp Tyr Met Phe Gln Phe His Arg Ile Leu Gln Tyr Ala

1910 1915 1920

Leu Pro Arg Gln Glu Ser Gln Arg Pro Phe Phe Trp Ile Phe Met

1925 1930 1935

Asp Asn Leu Leu Leu Thr Glu Asp Asp Gln Glu Thr Thr Thr Arg

1940 1945 1950

Phe Leu Gln Thr Glu Ala Val Thr Leu Gln Asp Val Arg Gly Arg

1955 1960 1965

Asp Tyr Gln Asn Ala Met Arg Val Trp Ser Asn Ile Pro Gly Leu

1970 1975 1980

Lys Ser Lys His Ala Pro Leu Thr Pro Lys Glu Glu Glu Tyr Leu

1985 1990 1995

Gln Ala Gln Val Arg Ser Arg Ser Lys Leu Asp Ala Pro Lys Val

2000 2005 2010

Asp Leu Leu Val Lys Asn Cys Leu Leu Pro Leu Arg Glu Tyr Phe

2015 2020 2025

Lys Tyr Phe Ser Gln Asn Ser Leu Pro Leu Ser Arg Ala Asp Pro

2030 2035 2040

Lys Lys Lys Arg Lys Val Gly Ser Gly Ala Thr Asn Phe Ser Leu

2045 2050 2055

Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Ser Glu

2060 2065 2070

Leu Ile Lys Glu Asn Met His Met Lys Leu Tyr Met Glu Gly Thr

2075 2080 2085

Val Asp Asn His His Phe Lys Cys Thr Ser Glu Gly Glu Gly Lys

2090 2095 2100

Pro Tyr Glu Gly Thr Gln Thr Met Arg Ile Lys Val Val Glu Gly

2105 2110 2115

Gly Pro Leu Pro Phe Ala Phe Asp Ile Leu Ala Thr Ser Phe Leu

2120 2125 2130

Tyr Gly Ser Lys Thr Phe Ile Asn His Thr Gln Gly Ile Pro Asp

2135 2140 2145

Phe Phe Lys Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg Val

2150 2155 2160

Thr Thr Tyr Glu Asp Gly Gly Val Leu Thr Ala Thr Gln Asp Thr

2165 2170 2175

Ser Leu Gln Asp Gly Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly

2180 2185 2190

Val Asn Phe Thr Ser Asn Gly Pro Val Met Gln Lys Lys Thr Leu

2195 2200 2205

Gly Trp Glu Ala Phe Thr Glu Thr Leu Tyr Pro Ala Asp Gly Gly

2210 2215 2220

Leu Glu Gly Arg Asn Asp Met Ala Leu Lys Leu Val Gly Gly Ser

2225 2230 2235

His Leu Ile Ala Asn Ile Lys Thr Thr Tyr Arg Ser Lys Lys Pro

2240 2245 2250

Ala Lys Asn Leu Lys Met Pro Gly Val Tyr Tyr Val Asp Tyr Arg

2255 2260 2265

Leu Glu Arg Ile Lys Glu Ala Asn Asn Glu Thr Tyr Val Glu Gln

2270 2275 2280

His Glu Val Ala Val Ala Arg Tyr Cys Asp Leu Pro Ser Lys Leu

2285 2290 2295

Gly His Lys Leu Asn

2300

<210> 8

<211> 2325

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 8

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Ser Gly Ser Glu Thr Pro Gly Thr Ser

65 70 75 80

Glu Ser Ala Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu

85 90 95

Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr

100 105 110

Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His

115 120 125

Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu

130 135 140

Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr

145 150 155 160

Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu

165 170 175

Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe

180 185 190

Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn

195 200 205

Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His

210 215 220

Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu

225 230 235 240

Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu

245 250 255

Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe

260 265 270

Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile

275 280 285

Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser

290 295 300

Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys

305 310 315 320

Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr

325 330 335

Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln

340 345 350

Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln

355 360 365

Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser

370 375 380

Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr

385 390 395 400

Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His

405 410 415

Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu

420 425 430

Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly

435 440 445

Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys

450 455 460

Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu

465 470 475 480

Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser

485 490 495

Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg

500 505 510

Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu

515 520 525

Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg

530 535 540

Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile

545 550 555 560

Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln

565 570 575

Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu

580 585 590

Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr

595 600 605

Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro

610 615 620

Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe

625 630 635 640

Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe

645 650 655

Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp

660 665 670

Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile

675 680 685

Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu

690 695 700

Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu

705 710 715 720

Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys

725 730 735

Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys

740 745 750

Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp

755 760 765

Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile

770 775 780

His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val

785 790 795 800

Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly

805 810 815

Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp

820 825 830

Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile

835 840 845

Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser

850 855 860

Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser

865 870 875 880

Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu

885 890 895

Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp

900 905 910

Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp Ala Ile

915 920 925

Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu

930 935 940

Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu

945 950 955 960

Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala

965 970 975

Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg

980 985 990

Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu

995 1000 1005

Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp

1010 1015 1020

Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg

1025 1030 1035

Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe

1040 1045 1050

Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr

1055 1060 1065

His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala

1070 1075 1080

Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly

1085 1090 1095

Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu

1100 1105 1110

Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn

1115 1120 1125

Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu

1130 1135 1140

Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu

1145 1150 1155

Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val

1160 1165 1170

Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln

1175 1180 1185

Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser

1190 1195 1200

Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr

1205 1210 1215

Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val

1220 1225 1230

Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys

1235 1240 1245

Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys

1250 1255 1260

Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys

1265 1270 1275

Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu Leu

1280 1285 1290

Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu Leu Gln

1295 1300 1305

Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn Phe Leu

1310 1315 1320

Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro Glu Asp

1325 1330 1335

Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His Tyr Leu

1340 1345 1350

Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg Val Ile

1355 1360 1365

Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys

1370 1375 1380

His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile Ile His

1385 1390 1395

Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys Tyr

1400 1405 1410

Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu

1415 1420 1425

Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr

1430 1435 1440

Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp Ser Arg Ala

1445 1450 1455

Asp Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Gly Ser Pro Lys

1460 1465 1470

Lys Lys Arg Lys Val Glu Ala Ser Gly Ser Gly Arg Ala Ser Pro

1475 1480 1485

Gly Ile Pro Gly Ser Thr Arg Asn His Asp Gln Glu Phe Asp Pro

1490 1495 1500

Pro Lys Val Tyr Pro Pro Val Pro Ala Glu Lys Arg Lys Pro Ile

1505 1510 1515

Arg Val Leu Ser Leu Phe Asp Gly Ile Ala Thr Gly Leu Leu Val

1520 1525 1530

Leu Lys Asp Leu Gly Ile Gln Val Asp Arg Tyr Ile Ala Ser Glu

1535 1540 1545

Val Cys Glu Asp Ser Ile Thr Val Gly Met Val Arg His Gln Gly

1550 1555 1560

Lys Ile Met Tyr Val Gly Asp Val Arg Ser Val Thr Gln Lys His

1565 1570 1575

Ile Gln Glu Trp Gly Pro Phe Asp Leu Val Ile Gly Gly Ser Pro

1580 1585 1590

Cys Asn Asp Leu Ser Ile Val Asn Pro Ala Arg Lys Gly Leu Tyr

1595 1600 1605

Glu Gly Thr Gly Arg Leu Phe Phe Glu Phe Tyr Arg Leu Leu His

1610 1615 1620

Asp Ala Arg Pro Lys Glu Gly Asp Asp Arg Pro Phe Phe Trp Leu

1625 1630 1635

Phe Glu Asn Val Val Ala Met Gly Val Ser Asp Lys Arg Asp Ile

1640 1645 1650

Ser Arg Phe Leu Glu Ser Asn Pro Val Met Ile Asp Ala Lys Glu

1655 1660 1665

Val Ser Ala Ala His Arg Ala Arg Tyr Phe Trp Gly Asn Leu Pro

1670 1675 1680

Gly Met Asn Arg Pro Leu Ala Ser Thr Val Asn Asp Lys Leu Glu

1685 1690 1695

Leu Gln Glu Cys Leu Glu His Gly Arg Ile Ala Lys Phe Ser Lys

1700 1705 1710

Val Arg Thr Ile Thr Thr Arg Ser Asn Ser Ile Lys Gln Gly Lys

1715 1720 1725

Asp Gln His Phe Pro Val Phe Met Asn Glu Lys Glu Asp Ile Leu

1730 1735 1740

Trp Cys Thr Glu Met Glu Arg Val Phe Gly Phe Pro Val His Tyr

1745 1750 1755

Thr Asp Val Ser Asn Met Ser Arg Leu Ala Arg Gln Arg Leu Leu

1760 1765 1770

Gly Arg Ser Trp Ser Val Pro Val Ile Arg His Leu Phe Ala Pro

1775 1780 1785

Leu Lys Glu Tyr Phe Ala Cys Val Ser Ser Gly Asn Ser Asn Ala

1790 1795 1800

Asn Ser Arg Gly Pro Ser Phe Ser Ser Gly Leu Val Pro Leu Ser

1805 1810 1815

Leu Arg Gly Ser His Met Gly Pro Met Glu Ile Tyr Lys Thr Val

1820 1825 1830

Ser Ala Trp Lys Arg Gln Pro Val Arg Val Leu Ser Leu Phe Arg

1835 1840 1845

Asn Ile Asp Lys Val Leu Lys Ser Leu Gly Phe Leu Glu Ser Gly

1850 1855 1860

Ser Gly Ser Gly Gly Gly Thr Leu Lys Tyr Val Glu Asp Val Thr

1865 1870 1875

Asn Val Val Arg Arg Asp Val Glu Lys Trp Gly Pro Phe Asp Leu

1880 1885 1890

Val Tyr Gly Ser Thr Gln Pro Leu Gly Ser Ser Cys Asp Arg Cys

1895 1900 1905

Pro Gly Trp Tyr Met Phe Gln Phe His Arg Ile Leu Gln Tyr Ala

1910 1915 1920

Leu Pro Arg Gln Glu Ser Gln Arg Pro Phe Phe Trp Ile Phe Met

1925 1930 1935

Asp Asn Leu Leu Leu Thr Glu Asp Asp Gln Glu Thr Thr Thr Arg

1940 1945 1950

Phe Leu Gln Thr Glu Ala Val Thr Leu Gln Asp Val Arg Gly Arg

1955 1960 1965

Asp Tyr Gln Asn Ala Met Arg Val Trp Ser Asn Ile Pro Gly Leu

1970 1975 1980

Lys Ser Lys His Ala Pro Leu Thr Pro Lys Glu Glu Glu Tyr Leu

1985 1990 1995

Gln Ala Gln Val Arg Ser Arg Ser Lys Leu Asp Ala Pro Lys Val

2000 2005 2010

Asp Leu Leu Val Lys Asn Cys Leu Leu Pro Leu Arg Glu Tyr Phe

2015 2020 2025

Lys Tyr Phe Ser Gln Asn Ser Leu Pro Leu Ser Arg Ala Asp Pro

2030 2035 2040

Lys Lys Lys Arg Lys Val Gly Ser Gly Ala Thr Asn Phe Ser Leu

2045 2050 2055

Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Gly Ser

2060 2065 2070

Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu

2075 2080 2085

Glu Asn Pro Gly Pro Ser Glu Leu Ile Lys Glu Asn Met His Met

2090 2095 2100

Lys Leu Tyr Met Glu Gly Thr Val Asp Asn His His Phe Lys Cys

2105 2110 2115

Thr Ser Glu Gly Glu Gly Lys Pro Tyr Glu Gly Thr Gln Thr Met

2120 2125 2130

Arg Ile Lys Val Val Glu Gly Gly Pro Leu Pro Phe Ala Phe Asp

2135 2140 2145

Ile Leu Ala Thr Ser Phe Leu Tyr Gly Ser Lys Thr Phe Ile Asn

2150 2155 2160

His Thr Gln Gly Ile Pro Asp Phe Phe Lys Gln Ser Phe Pro Glu

2165 2170 2175

Gly Phe Thr Trp Glu Arg Val Thr Thr Tyr Glu Asp Gly Gly Val

2180 2185 2190

Leu Thr Ala Thr Gln Asp Thr Ser Leu Gln Asp Gly Cys Leu Ile

2195 2200 2205

Tyr Asn Val Lys Ile Arg Gly Val Asn Phe Thr Ser Asn Gly Pro

2210 2215 2220

Val Met Gln Lys Lys Thr Leu Gly Trp Glu Ala Phe Thr Glu Thr

2225 2230 2235

Leu Tyr Pro Ala Asp Gly Gly Leu Glu Gly Arg Asn Asp Met Ala

2240 2245 2250

Leu Lys Leu Val Gly Gly Ser His Leu Ile Ala Asn Ile Lys Thr

2255 2260 2265

Thr Tyr Arg Ser Lys Lys Pro Ala Lys Asn Leu Lys Met Pro Gly

2270 2275 2280

Val Tyr Tyr Val Asp Tyr Arg Leu Glu Arg Ile Lys Glu Ala Asn

2285 2290 2295

Asn Glu Thr Tyr Val Glu Gln His Glu Val Ala Val Ala Arg Tyr

2300 2305 2310

Cys Asp Leu Pro Ser Lys Leu Gly His Lys Leu Asn

2315 2320 2325

<210> 9

<211> 2367

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 9

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Gly Gly Pro Ser Ser Gly Ala Pro Pro

65 70 75 80

Pro Ser Gly Gly Ser Pro Ala Gly Ser Pro Thr Ser Thr Glu Glu Gly

85 90 95

Thr Ser Glu Ser Ala Thr Pro Glu Ser Gly Pro Gly Thr Ser Thr Glu

100 105 110

Pro Ser Glu Gly Ser Ala Pro Gly Ser Pro Ala Gly Ser Pro Thr Ser

115 120 125

Thr Glu Glu Gly Thr Ser Thr Glu Pro Ser Glu Gly Ser Ala Pro Gly

130 135 140

Thr Ser Thr Glu Pro Ser Glu Met Asp Lys Lys Tyr Ser Ile Gly Leu

145 150 155 160

Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr

165 170 175

Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His

180 185 190

Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu

195 200 205

Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr

210 215 220

Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu

225 230 235 240

Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe

245 250 255

Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn

260 265 270

Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His

275 280 285

Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu

290 295 300

Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu

305 310 315 320

Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe

325 330 335

Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile

340 345 350

Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser

355 360 365

Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys

370 375 380

Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr

385 390 395 400

Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln

405 410 415

Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln

420 425 430

Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser

435 440 445

Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr

450 455 460

Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His

465 470 475 480

Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu

485 490 495

Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly

500 505 510

Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys

515 520 525

Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu

530 535 540

Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser

545 550 555 560

Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg

565 570 575

Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu

580 585 590

Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg

595 600 605

Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile

610 615 620

Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln

625 630 635 640

Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu

645 650 655

Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr

660 665 670

Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro

675 680 685

Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe

690 695 700

Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe

705 710 715 720

Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp

725 730 735

Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile

740 745 750

Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu

755 760 765

Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu

770 775 780

Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys

785 790 795 800

Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys

805 810 815

Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp

820 825 830

Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile

835 840 845

His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val

850 855 860

Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly

865 870 875 880

Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp

885 890 895

Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile

900 905 910

Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser

915 920 925

Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser

930 935 940

Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu

945 950 955 960

Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp

965 970 975

Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp Ala Ile

980 985 990

Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu

995 1000 1005

Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser

1010 1015 1020

Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu

1025 1030 1035

Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys

1040 1045 1050

Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile

1055 1060 1065

Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala

1070 1075 1080

Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp

1085 1090 1095

Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu

1100 1105 1110

Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu

1115 1120 1125

Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val

1130 1135 1140

Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu

1145 1150 1155

Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile

1160 1165 1170

Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe

1175 1180 1185

Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu

1190 1195 1200

Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly

1205 1210 1215

Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr

1220 1225 1230

Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys

1235 1240 1245

Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro

1250 1255 1260

Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp

1265 1270 1275

Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser

1280 1285 1290

Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu

1295 1300 1305

Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser

1310 1315 1320

Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr

1325 1330 1335

Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser

1340 1345 1350

Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala

1355 1360 1365

Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr

1370 1375 1380

Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly

1385 1390 1395

Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His

1400 1405 1410

Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser

1415 1420 1425

Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser

1430 1435 1440

Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu

1445 1450 1455

Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala

1460 1465 1470

Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr

1475 1480 1485

Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile

1490 1495 1500

Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly

1505 1510 1515

Asp Ser Arg Ala Asp Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser

1520 1525 1530

Gly Ser Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly Ser Gly

1535 1540 1545

Arg Ala Ser Pro Gly Ile Pro Gly Ser Thr Arg Asn His Asp Gln

1550 1555 1560

Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro Ala Glu Lys

1565 1570 1575

Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile Ala Thr

1580 1585 1590

Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp Arg Tyr

1595 1600 1605

Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly Met Val

1610 1615 1620

Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg Ser Val

1625 1630 1635

Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu Val Ile

1640 1645 1650

Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro Ala Arg

1655 1660 1665

Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu Phe Tyr

1670 1675 1680

Arg Leu Leu His Asp Ala Arg Pro Lys Glu Gly Asp Asp Arg Pro

1685 1690 1695

Phe Phe Trp Leu Phe Glu Asn Val Val Ala Met Gly Val Ser Asp

1700 1705 1710

Lys Arg Asp Ile Ser Arg Phe Leu Glu Ser Asn Pro Val Met Ile

1715 1720 1725

Asp Ala Lys Glu Val Ser Ala Ala His Arg Ala Arg Tyr Phe Trp

1730 1735 1740

Gly Asn Leu Pro Gly Met Asn Arg Pro Leu Ala Ser Thr Val Asn

1745 1750 1755

Asp Lys Leu Glu Leu Gln Glu Cys Leu Glu His Gly Arg Ile Ala

1760 1765 1770

Lys Phe Ser Lys Val Arg Thr Ile Thr Thr Arg Ser Asn Ser Ile

1775 1780 1785

Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe Met Asn Glu Lys

1790 1795 1800

Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val Phe Gly Phe

1805 1810 1815

Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu Ala Arg

1820 1825 1830

Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile Arg His

1835 1840 1845

Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val Ser Ser Gly

1850 1855 1860

Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser Gly Leu

1865 1870 1875

Val Pro Leu Ser Leu Arg Gly Ser His Met Gly Pro Met Glu Ile

1880 1885 1890

Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln Pro Val Arg Val Leu

1895 1900 1905

Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys Ser Leu Gly Phe

1910 1915 1920

Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr Leu Lys Tyr Val

1925 1930 1935

Glu Asp Val Thr Asn Val Val Arg Arg Asp Val Glu Lys Trp Gly

1940 1945 1950

Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro Leu Gly Ser Ser

1955 1960 1965

Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln Phe His Arg Ile

1970 1975 1980

Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln Arg Pro Phe Phe

1985 1990 1995

Trp Ile Phe Met Asp Asn Leu Leu Leu Thr Glu Asp Asp Gln Glu

2000 2005 2010

Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala Val Thr Leu Gln Asp

2015 2020 2025

Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg Val Trp Ser Asn

2030 2035 2040

Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu Thr Pro Lys Glu

2045 2050 2055

Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg Ser Lys Leu Asp

2060 2065 2070

Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys Leu Leu Pro Leu

2075 2080 2085

Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser Leu Pro Leu Ser

2090 2095 2100

Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Gly Ser Gly Ala Thr

2105 2110 2115

Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro

2120 2125 2130

Gly Pro Ser Glu Leu Ile Lys Glu Asn Met His Met Lys Leu Tyr

2135 2140 2145

Met Glu Gly Thr Val Asp Asn His His Phe Lys Cys Thr Ser Glu

2150 2155 2160

Gly Glu Gly Lys Pro Tyr Glu Gly Thr Gln Thr Met Arg Ile Lys

2165 2170 2175

Val Val Glu Gly Gly Pro Leu Pro Phe Ala Phe Asp Ile Leu Ala

2180 2185 2190

Thr Ser Phe Leu Tyr Gly Ser Lys Thr Phe Ile Asn His Thr Gln

2195 2200 2205

Gly Ile Pro Asp Phe Phe Lys Gln Ser Phe Pro Glu Gly Phe Thr

2210 2215 2220

Trp Glu Arg Val Thr Thr Tyr Glu Asp Gly Gly Val Leu Thr Ala

2225 2230 2235

Thr Gln Asp Thr Ser Leu Gln Asp Gly Cys Leu Ile Tyr Asn Val

2240 2245 2250

Lys Ile Arg Gly Val Asn Phe Thr Ser Asn Gly Pro Val Met Gln

2255 2260 2265

Lys Lys Thr Leu Gly Trp Glu Ala Phe Thr Glu Thr Leu Tyr Pro

2270 2275 2280

Ala Asp Gly Gly Leu Glu Gly Arg Asn Asp Met Ala Leu Lys Leu

2285 2290 2295

Val Gly Gly Ser His Leu Ile Ala Asn Ile Lys Thr Thr Tyr Arg

2300 2305 2310

Ser Lys Lys Pro Ala Lys Asn Leu Lys Met Pro Gly Val Tyr Tyr

2315 2320 2325

Val Asp Tyr Arg Leu Glu Arg Ile Lys Glu Ala Asn Asn Glu Thr

2330 2335 2340

Tyr Val Glu Gln His Glu Val Ala Val Ala Arg Tyr Cys Asp Leu

2345 2350 2355

Pro Ser Lys Leu Gly His Lys Leu Asn

2360 2365

<210> 10

<211> 2389

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 10

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Gly Gly Pro Ser Ser Gly Ala Pro Pro

65 70 75 80

Pro Ser Gly Gly Ser Pro Ala Gly Ser Pro Thr Ser Thr Glu Glu Gly

85 90 95

Thr Ser Glu Ser Ala Thr Pro Glu Ser Gly Pro Gly Thr Ser Thr Glu

100 105 110

Pro Ser Glu Gly Ser Ala Pro Gly Ser Pro Ala Gly Ser Pro Thr Ser

115 120 125

Thr Glu Glu Gly Thr Ser Thr Glu Pro Ser Glu Gly Ser Ala Pro Gly

130 135 140

Thr Ser Thr Glu Pro Ser Glu Met Asp Lys Lys Tyr Ser Ile Gly Leu

145 150 155 160

Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr

165 170 175

Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His

180 185 190

Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu

195 200 205

Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr

210 215 220

Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu

225 230 235 240

Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe

245 250 255

Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn

260 265 270

Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His

275 280 285

Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu

290 295 300

Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu

305 310 315 320

Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe

325 330 335

Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile

340 345 350

Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser

355 360 365

Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys

370 375 380

Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr

385 390 395 400

Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln

405 410 415

Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln

420 425 430

Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser

435 440 445

Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr

450 455 460

Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His

465 470 475 480

Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu

485 490 495

Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly

500 505 510

Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys

515 520 525

Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu

530 535 540

Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser

545 550 555 560

Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg

565 570 575

Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu

580 585 590

Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg

595 600 605

Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile

610 615 620

Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln

625 630 635 640

Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu

645 650 655

Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr

660 665 670

Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro

675 680 685

Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe

690 695 700

Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe

705 710 715 720

Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp

725 730 735

Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile

740 745 750

Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu

755 760 765

Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu

770 775 780

Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys

785 790 795 800

Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys

805 810 815

Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp

820 825 830

Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile

835 840 845

His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val

850 855 860

Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly

865 870 875 880

Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp

885 890 895

Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile

900 905 910

Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser

915 920 925

Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser

930 935 940

Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu

945 950 955 960

Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp

965 970 975

Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp Ala Ile

980 985 990

Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu

995 1000 1005

Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser

1010 1015 1020

Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu

1025 1030 1035

Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys

1040 1045 1050

Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile

1055 1060 1065

Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala

1070 1075 1080

Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp

1085 1090 1095

Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu

1100 1105 1110

Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu

1115 1120 1125

Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val

1130 1135 1140

Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu

1145 1150 1155

Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile

1160 1165 1170

Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe

1175 1180 1185

Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu

1190 1195 1200

Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly

1205 1210 1215

Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr

1220 1225 1230

Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys

1235 1240 1245

Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro

1250 1255 1260

Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp

1265 1270 1275

Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser

1280 1285 1290

Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu

1295 1300 1305

Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser

1310 1315 1320

Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr

1325 1330 1335

Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser

1340 1345 1350

Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala

1355 1360 1365

Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr

1370 1375 1380

Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly

1385 1390 1395

Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His

1400 1405 1410

Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser

1415 1420 1425

Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser

1430 1435 1440

Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu

1445 1450 1455

Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala

1460 1465 1470

Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr

1475 1480 1485

Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile

1490 1495 1500

Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly

1505 1510 1515

Asp Ser Arg Ala Asp Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser

1520 1525 1530

Gly Ser Pro Lys Lys Lys Arg Lys Val Glu Ala Ser Gly Ser Gly

1535 1540 1545

Arg Ala Ser Pro Gly Ile Pro Gly Ser Thr Arg Asn His Asp Gln

1550 1555 1560

Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro Ala Glu Lys

1565 1570 1575

Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile Ala Thr

1580 1585 1590

Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp Arg Tyr

1595 1600 1605

Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly Met Val

1610 1615 1620

Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg Ser Val

1625 1630 1635

Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu Val Ile

1640 1645 1650

Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro Ala Arg

1655 1660 1665

Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu Phe Tyr

1670 1675 1680

Arg Leu Leu His Asp Ala Arg Pro Lys Glu Gly Asp Asp Arg Pro

1685 1690 1695

Phe Phe Trp Leu Phe Glu Asn Val Val Ala Met Gly Val Ser Asp

1700 1705 1710

Lys Arg Asp Ile Ser Arg Phe Leu Glu Ser Asn Pro Val Met Ile

1715 1720 1725

Asp Ala Lys Glu Val Ser Ala Ala His Arg Ala Arg Tyr Phe Trp

1730 1735 1740

Gly Asn Leu Pro Gly Met Asn Arg Pro Leu Ala Ser Thr Val Asn

1745 1750 1755

Asp Lys Leu Glu Leu Gln Glu Cys Leu Glu His Gly Arg Ile Ala

1760 1765 1770

Lys Phe Ser Lys Val Arg Thr Ile Thr Thr Arg Ser Asn Ser Ile

1775 1780 1785

Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe Met Asn Glu Lys

1790 1795 1800

Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val Phe Gly Phe

1805 1810 1815

Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu Ala Arg

1820 1825 1830

Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile Arg His

1835 1840 1845

Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val Ser Ser Gly

1850 1855 1860

Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser Gly Leu

1865 1870 1875

Val Pro Leu Ser Leu Arg Gly Ser His Met Gly Pro Met Glu Ile

1880 1885 1890

Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln Pro Val Arg Val Leu

1895 1900 1905

Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys Ser Leu Gly Phe

1910 1915 1920

Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr Leu Lys Tyr Val

1925 1930 1935

Glu Asp Val Thr Asn Val Val Arg Arg Asp Val Glu Lys Trp Gly

1940 1945 1950

Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro Leu Gly Ser Ser

1955 1960 1965

Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln Phe His Arg Ile

1970 1975 1980

Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln Arg Pro Phe Phe

1985 1990 1995

Trp Ile Phe Met Asp Asn Leu Leu Leu Thr Glu Asp Asp Gln Glu

2000 2005 2010

Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala Val Thr Leu Gln Asp

2015 2020 2025

Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg Val Trp Ser Asn

2030 2035 2040

Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu Thr Pro Lys Glu

2045 2050 2055

Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg Ser Lys Leu Asp

2060 2065 2070

Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys Leu Leu Pro Leu

2075 2080 2085

Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser Leu Pro Leu Ser

2090 2095 2100

Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Gly Ser Gly Ala Thr

2105 2110 2115

Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn Pro

2120 2125 2130

Gly Pro Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala

2135 2140 2145

Gly Asp Val Glu Glu Asn Pro Gly Pro Ser Glu Leu Ile Lys Glu

2150 2155 2160

Asn Met His Met Lys Leu Tyr Met Glu Gly Thr Val Asp Asn His

2165 2170 2175

His Phe Lys Cys Thr Ser Glu Gly Glu Gly Lys Pro Tyr Glu Gly

2180 2185 2190

Thr Gln Thr Met Arg Ile Lys Val Val Glu Gly Gly Pro Leu Pro

2195 2200 2205

Phe Ala Phe Asp Ile Leu Ala Thr Ser Phe Leu Tyr Gly Ser Lys

2210 2215 2220

Thr Phe Ile Asn His Thr Gln Gly Ile Pro Asp Phe Phe Lys Gln

2225 2230 2235

Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg Val Thr Thr Tyr Glu

2240 2245 2250

Asp Gly Gly Val Leu Thr Ala Thr Gln Asp Thr Ser Leu Gln Asp

2255 2260 2265

Gly Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly Val Asn Phe Thr

2270 2275 2280

Ser Asn Gly Pro Val Met Gln Lys Lys Thr Leu Gly Trp Glu Ala

2285 2290 2295

Phe Thr Glu Thr Leu Tyr Pro Ala Asp Gly Gly Leu Glu Gly Arg

2300 2305 2310

Asn Asp Met Ala Leu Lys Leu Val Gly Gly Ser His Leu Ile Ala

2315 2320 2325

Asn Ile Lys Thr Thr Tyr Arg Ser Lys Lys Pro Ala Lys Asn Leu

2330 2335 2340

Lys Met Pro Gly Val Tyr Tyr Val Asp Tyr Arg Leu Glu Arg Ile

2345 2350 2355

Lys Glu Ala Asn Asn Glu Thr Tyr Val Glu Gln His Glu Val Ala

2360 2365 2370

Val Ala Arg Tyr Cys Asp Leu Pro Ser Lys Leu Gly His Lys Leu

2375 2380 2385

Asn

<210> 11

<211> 2369

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 11

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Ser Gly Ser Glu Thr Pro Gly Thr Ser

65 70 75 80

Glu Ser Ala Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu

85 90 95

Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr

100 105 110

Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His

115 120 125

Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu

130 135 140

Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr

145 150 155 160

Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu

165 170 175

Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe

180 185 190

Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn

195 200 205

Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His

210 215 220

Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu

225 230 235 240

Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu

245 250 255

Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe

260 265 270

Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile

275 280 285

Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser

290 295 300

Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys

305 310 315 320

Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr

325 330 335

Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln

340 345 350

Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln

355 360 365

Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser

370 375 380

Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr

385 390 395 400

Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His

405 410 415

Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu

420 425 430

Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly

435 440 445

Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys

450 455 460

Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu

465 470 475 480

Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser

485 490 495

Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg

500 505 510

Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu

515 520 525

Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg

530 535 540

Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile

545 550 555 560

Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln

565 570 575

Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu

580 585 590

Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr

595 600 605

Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro

610 615 620

Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe

625 630 635 640

Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe

645 650 655

Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp

660 665 670

Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile

675 680 685

Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu

690 695 700

Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu

705 710 715 720

Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys

725 730 735

Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys

740 745 750

Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp

755 760 765

Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile

770 775 780

His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val

785 790 795 800

Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly

805 810 815

Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp

820 825 830

Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile

835 840 845

Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser

850 855 860

Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser

865 870 875 880

Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu

885 890 895

Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp

900 905 910

Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp Ala Ile

915 920 925

Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu

930 935 940

Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu

945 950 955 960

Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala

965 970 975

Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg

980 985 990

Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu

995 1000 1005

Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp

1010 1015 1020

Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg

1025 1030 1035

Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe

1040 1045 1050

Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr

1055 1060 1065

His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala

1070 1075 1080

Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly

1085 1090 1095

Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu

1100 1105 1110

Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn

1115 1120 1125

Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu

1130 1135 1140

Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu

1145 1150 1155

Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val

1160 1165 1170

Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln

1175 1180 1185

Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser

1190 1195 1200

Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr

1205 1210 1215

Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val

1220 1225 1230

Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys

1235 1240 1245

Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys

1250 1255 1260

Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys

1265 1270 1275

Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu Leu

1280 1285 1290

Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu Leu Gln

1295 1300 1305

Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn Phe Leu

1310 1315 1320

Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro Glu Asp

1325 1330 1335

Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His Tyr Leu

1340 1345 1350

Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg Val Ile

1355 1360 1365

Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys

1370 1375 1380

His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile Ile His

1385 1390 1395

Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys Tyr

1400 1405 1410

Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu

1415 1420 1425

Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr

1430 1435 1440

Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp Ser Arg Ala

1445 1450 1455

Asp Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Gly Ser Pro Lys

1460 1465 1470

Lys Lys Arg Lys Val Ser Pro Gly Gly Gly Pro Ser Ser Gly Ala

1475 1480 1485

Pro Pro Pro Ser Gly Gly Ser Pro Ala Gly Ser Pro Thr Ser Thr

1490 1495 1500

Glu Glu Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Gly Pro Gly

1505 1510 1515

Thr Ser Thr Glu Pro Ser Glu Gly Ser Ala Pro Gly Ser Pro Ala

1520 1525 1530

Gly Ser Pro Thr Ser Thr Glu Glu Gly Thr Ser Thr Glu Pro Ser

1535 1540 1545

Glu Gly Ser Ala Pro Gly Thr Ser Thr Glu Pro Ser Glu Asn His

1550 1555 1560

Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro Ala

1565 1570 1575

Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile

1580 1585 1590

Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp

1595 1600 1605

Arg Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly

1610 1615 1620

Met Val Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg

1625 1630 1635

Ser Val Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu

1640 1645 1650

Val Ile Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro

1655 1660 1665

Ala Arg Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu

1670 1675 1680

Phe Tyr Arg Leu Leu His Asp Ala Arg Pro Lys Glu Gly Asp Asp

1685 1690 1695

Arg Pro Phe Phe Trp Leu Phe Glu Asn Val Val Ala Met Gly Val

1700 1705 1710

Ser Asp Lys Arg Asp Ile Ser Arg Phe Leu Glu Ser Asn Pro Val

1715 1720 1725

Met Ile Asp Ala Lys Glu Val Ser Ala Ala His Arg Ala Arg Tyr

1730 1735 1740

Phe Trp Gly Asn Leu Pro Gly Met Asn Arg Pro Leu Ala Ser Thr

1745 1750 1755

Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu Glu His Gly Arg

1760 1765 1770

Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr Arg Ser Asn

1775 1780 1785

Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe Met Asn

1790 1795 1800

Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val Phe

1805 1810 1815

Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu

1820 1825 1830

Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile

1835 1840 1845

Arg His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val Ser

1850 1855 1860

Ser Gly Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser

1865 1870 1875

Gly Leu Val Pro Leu Ser Leu Arg Gly Ser His Met Gly Pro Met

1880 1885 1890

Glu Ile Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln Pro Val Arg

1895 1900 1905

Val Leu Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys Ser Leu

1910 1915 1920

Gly Phe Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr Leu Lys

1925 1930 1935

Tyr Val Glu Asp Val Thr Asn Val Val Arg Arg Asp Val Glu Lys

1940 1945 1950

Trp Gly Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro Leu Gly

1955 1960 1965

Ser Ser Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln Phe His

1970 1975 1980

Arg Ile Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln Arg Pro

1985 1990 1995

Phe Phe Trp Ile Phe Met Asp Asn Leu Leu Leu Thr Glu Asp Asp

2000 2005 2010

Gln Glu Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala Val Thr Leu

2015 2020 2025

Gln Asp Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg Val Trp

2030 2035 2040

Ser Asn Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu Thr Pro

2045 2050 2055

Lys Glu Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg Ser Lys

2060 2065 2070

Leu Asp Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys Leu Leu

2075 2080 2085

Pro Leu Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser Leu Pro

2090 2095 2100

Leu Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Gly Ser Gly

2105 2110 2115

Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu

2120 2125 2130

Asn Pro Gly Pro Ser Glu Leu Ile Lys Glu Asn Met His Met Lys

2135 2140 2145

Leu Tyr Met Glu Gly Thr Val Asp Asn His His Phe Lys Cys Thr

2150 2155 2160

Ser Glu Gly Glu Gly Lys Pro Tyr Glu Gly Thr Gln Thr Met Arg

2165 2170 2175

Ile Lys Val Val Glu Gly Gly Pro Leu Pro Phe Ala Phe Asp Ile

2180 2185 2190

Leu Ala Thr Ser Phe Leu Tyr Gly Ser Lys Thr Phe Ile Asn His

2195 2200 2205

Thr Gln Gly Ile Pro Asp Phe Phe Lys Gln Ser Phe Pro Glu Gly

2210 2215 2220

Phe Thr Trp Glu Arg Val Thr Thr Tyr Glu Asp Gly Gly Val Leu

2225 2230 2235

Thr Ala Thr Gln Asp Thr Ser Leu Gln Asp Gly Cys Leu Ile Tyr

2240 2245 2250

Asn Val Lys Ile Arg Gly Val Asn Phe Thr Ser Asn Gly Pro Val

2255 2260 2265

Met Gln Lys Lys Thr Leu Gly Trp Glu Ala Phe Thr Glu Thr Leu

2270 2275 2280

Tyr Pro Ala Asp Gly Gly Leu Glu Gly Arg Asn Asp Met Ala Leu

2285 2290 2295

Lys Leu Val Gly Gly Ser His Leu Ile Ala Asn Ile Lys Thr Thr

2300 2305 2310

Tyr Arg Ser Lys Lys Pro Ala Lys Asn Leu Lys Met Pro Gly Val

2315 2320 2325

Tyr Tyr Val Asp Tyr Arg Leu Glu Arg Ile Lys Glu Ala Asn Asn

2330 2335 2340

Glu Thr Tyr Val Glu Gln His Glu Val Ala Val Ala Arg Tyr Cys

2345 2350 2355

Asp Leu Pro Ser Lys Leu Gly His Lys Leu Asn

2360 2365

<210> 12

<211> 2391

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 12

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Ser Gly Ser Glu Thr Pro Gly Thr Ser

65 70 75 80

Glu Ser Ala Thr Pro Glu Ser Met Asp Lys Lys Tyr Ser Ile Gly Leu

85 90 95

Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr

100 105 110

Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His

115 120 125

Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu

130 135 140

Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr

145 150 155 160

Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu

165 170 175

Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe

180 185 190

Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn

195 200 205

Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His

210 215 220

Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu

225 230 235 240

Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu

245 250 255

Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe

260 265 270

Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile

275 280 285

Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser

290 295 300

Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys

305 310 315 320

Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr

325 330 335

Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln

340 345 350

Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln

355 360 365

Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser

370 375 380

Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr

385 390 395 400

Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His

405 410 415

Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu

420 425 430

Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly

435 440 445

Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys

450 455 460

Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu

465 470 475 480

Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser

485 490 495

Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg

500 505 510

Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu

515 520 525

Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg

530 535 540

Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile

545 550 555 560

Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln

565 570 575

Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu

580 585 590

Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr

595 600 605

Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro

610 615 620

Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe

625 630 635 640

Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe

645 650 655

Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp

660 665 670

Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile

675 680 685

Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu

690 695 700

Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu

705 710 715 720

Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys

725 730 735

Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys

740 745 750

Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp

755 760 765

Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile

770 775 780

His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val

785 790 795 800

Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly

805 810 815

Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp

820 825 830

Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile

835 840 845

Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser

850 855 860

Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser

865 870 875 880

Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu

885 890 895

Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp

900 905 910

Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp Ala Ile

915 920 925

Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu

930 935 940

Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu

945 950 955 960

Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala

965 970 975

Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg

980 985 990

Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu

995 1000 1005

Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp

1010 1015 1020

Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg

1025 1030 1035

Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe

1040 1045 1050

Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr

1055 1060 1065

His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala

1070 1075 1080

Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly

1085 1090 1095

Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu

1100 1105 1110

Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn

1115 1120 1125

Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu

1130 1135 1140

Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu

1145 1150 1155

Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val

1160 1165 1170

Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln

1175 1180 1185

Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser

1190 1195 1200

Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr

1205 1210 1215

Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val

1220 1225 1230

Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys

1235 1240 1245

Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys

1250 1255 1260

Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys

1265 1270 1275

Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu Leu

1280 1285 1290

Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu Leu Gln

1295 1300 1305

Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn Phe Leu

1310 1315 1320

Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro Glu Asp

1325 1330 1335

Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His Tyr Leu

1340 1345 1350

Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg Val Ile

1355 1360 1365

Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys

1370 1375 1380

His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile Ile His

1385 1390 1395

Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys Tyr

1400 1405 1410

Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu

1415 1420 1425

Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr

1430 1435 1440

Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp Ser Arg Ala

1445 1450 1455

Asp Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Gly Ser Pro Lys

1460 1465 1470

Lys Lys Arg Lys Val Ser Pro Gly Gly Gly Pro Ser Ser Gly Ala

1475 1480 1485

Pro Pro Pro Ser Gly Gly Ser Pro Ala Gly Ser Pro Thr Ser Thr

1490 1495 1500

Glu Glu Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Gly Pro Gly

1505 1510 1515

Thr Ser Thr Glu Pro Ser Glu Gly Ser Ala Pro Gly Ser Pro Ala

1520 1525 1530

Gly Ser Pro Thr Ser Thr Glu Glu Gly Thr Ser Thr Glu Pro Ser

1535 1540 1545

Glu Gly Ser Ala Pro Gly Thr Ser Thr Glu Pro Ser Glu Asn His

1550 1555 1560

Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro Ala

1565 1570 1575

Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile

1580 1585 1590

Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp

1595 1600 1605

Arg Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly

1610 1615 1620

Met Val Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg

1625 1630 1635

Ser Val Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu

1640 1645 1650

Val Ile Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro

1655 1660 1665

Ala Arg Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu

1670 1675 1680

Phe Tyr Arg Leu Leu His Asp Ala Arg Pro Lys Glu Gly Asp Asp

1685 1690 1695

Arg Pro Phe Phe Trp Leu Phe Glu Asn Val Val Ala Met Gly Val

1700 1705 1710

Ser Asp Lys Arg Asp Ile Ser Arg Phe Leu Glu Ser Asn Pro Val

1715 1720 1725

Met Ile Asp Ala Lys Glu Val Ser Ala Ala His Arg Ala Arg Tyr

1730 1735 1740

Phe Trp Gly Asn Leu Pro Gly Met Asn Arg Pro Leu Ala Ser Thr

1745 1750 1755

Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu Glu His Gly Arg

1760 1765 1770

Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr Arg Ser Asn

1775 1780 1785

Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe Met Asn

1790 1795 1800

Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val Phe

1805 1810 1815

Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu

1820 1825 1830

Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile

1835 1840 1845

Arg His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val Ser

1850 1855 1860

Ser Gly Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser

1865 1870 1875

Gly Leu Val Pro Leu Ser Leu Arg Gly Ser His Met Gly Pro Met

1880 1885 1890

Glu Ile Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln Pro Val Arg

1895 1900 1905

Val Leu Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys Ser Leu

1910 1915 1920

Gly Phe Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr Leu Lys

1925 1930 1935

Tyr Val Glu Asp Val Thr Asn Val Val Arg Arg Asp Val Glu Lys

1940 1945 1950

Trp Gly Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro Leu Gly

1955 1960 1965

Ser Ser Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln Phe His

1970 1975 1980

Arg Ile Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln Arg Pro

1985 1990 1995

Phe Phe Trp Ile Phe Met Asp Asn Leu Leu Leu Thr Glu Asp Asp

2000 2005 2010

Gln Glu Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala Val Thr Leu

2015 2020 2025

Gln Asp Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg Val Trp

2030 2035 2040

Ser Asn Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu Thr Pro

2045 2050 2055

Lys Glu Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg Ser Lys

2060 2065 2070

Leu Asp Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys Leu Leu

2075 2080 2085

Pro Leu Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser Leu Pro

2090 2095 2100

Leu Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Gly Ser Gly

2105 2110 2115

Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu

2120 2125 2130

Asn Pro Gly Pro Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys

2135 2140 2145

Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Ser Glu Leu Ile

2150 2155 2160

Lys Glu Asn Met His Met Lys Leu Tyr Met Glu Gly Thr Val Asp

2165 2170 2175

Asn His His Phe Lys Cys Thr Ser Glu Gly Glu Gly Lys Pro Tyr

2180 2185 2190

Glu Gly Thr Gln Thr Met Arg Ile Lys Val Val Glu Gly Gly Pro

2195 2200 2205

Leu Pro Phe Ala Phe Asp Ile Leu Ala Thr Ser Phe Leu Tyr Gly

2210 2215 2220

Ser Lys Thr Phe Ile Asn His Thr Gln Gly Ile Pro Asp Phe Phe

2225 2230 2235

Lys Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg Val Thr Thr

2240 2245 2250

Tyr Glu Asp Gly Gly Val Leu Thr Ala Thr Gln Asp Thr Ser Leu

2255 2260 2265

Gln Asp Gly Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly Val Asn

2270 2275 2280

Phe Thr Ser Asn Gly Pro Val Met Gln Lys Lys Thr Leu Gly Trp

2285 2290 2295

Glu Ala Phe Thr Glu Thr Leu Tyr Pro Ala Asp Gly Gly Leu Glu

2300 2305 2310

Gly Arg Asn Asp Met Ala Leu Lys Leu Val Gly Gly Ser His Leu

2315 2320 2325

Ile Ala Asn Ile Lys Thr Thr Tyr Arg Ser Lys Lys Pro Ala Lys

2330 2335 2340

Asn Leu Lys Met Pro Gly Val Tyr Tyr Val Asp Tyr Arg Leu Glu

2345 2350 2355

Arg Ile Lys Glu Ala Asn Asn Glu Thr Tyr Val Glu Gln His Glu

2360 2365 2370

Val Ala Val Ala Arg Tyr Cys Asp Leu Pro Ser Lys Leu Gly His

2375 2380 2385

Lys Leu Asn

2390

<210> 13

<211> 2369

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 13

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Gly Gly Pro Ser Ser Gly Ala Pro Pro

65 70 75 80

Pro Ser Gly Gly Ser Pro Ala Gly Ser Pro Thr Ser Thr Glu Glu Gly

85 90 95

Thr Ser Glu Ser Ala Thr Pro Glu Ser Gly Pro Gly Thr Ser Thr Glu

100 105 110

Pro Ser Glu Gly Ser Ala Pro Gly Ser Pro Ala Gly Ser Pro Thr Ser

115 120 125

Thr Glu Glu Gly Thr Ser Thr Glu Pro Ser Glu Gly Ser Ala Pro Gly

130 135 140

Thr Ser Thr Glu Pro Ser Glu Met Asp Lys Lys Tyr Ser Ile Gly Leu

145 150 155 160

Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr

165 170 175

Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His

180 185 190

Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu

195 200 205

Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr

210 215 220

Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu

225 230 235 240

Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe

245 250 255

Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn

260 265 270

Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His

275 280 285

Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu

290 295 300

Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu

305 310 315 320

Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe

325 330 335

Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile

340 345 350

Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser

355 360 365

Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys

370 375 380

Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr

385 390 395 400

Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln

405 410 415

Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln

420 425 430

Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser

435 440 445

Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr

450 455 460

Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His

465 470 475 480

Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu

485 490 495

Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly

500 505 510

Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys

515 520 525

Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu

530 535 540

Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser

545 550 555 560

Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg

565 570 575

Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu

580 585 590

Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg

595 600 605

Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile

610 615 620

Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln

625 630 635 640

Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu

645 650 655

Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr

660 665 670

Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro

675 680 685

Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe

690 695 700

Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe

705 710 715 720

Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp

725 730 735

Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile

740 745 750

Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu

755 760 765

Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu

770 775 780

Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys

785 790 795 800

Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys

805 810 815

Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp

820 825 830

Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile

835 840 845

His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val

850 855 860

Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly

865 870 875 880

Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp

885 890 895

Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile

900 905 910

Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser

915 920 925

Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser

930 935 940

Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu

945 950 955 960

Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp

965 970 975

Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp Ala Ile

980 985 990

Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu

995 1000 1005

Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser

1010 1015 1020

Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu

1025 1030 1035

Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys

1040 1045 1050

Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile

1055 1060 1065

Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala

1070 1075 1080

Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp

1085 1090 1095

Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu

1100 1105 1110

Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu

1115 1120 1125

Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val

1130 1135 1140

Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu

1145 1150 1155

Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile

1160 1165 1170

Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe

1175 1180 1185

Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu

1190 1195 1200

Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly

1205 1210 1215

Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr

1220 1225 1230

Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys

1235 1240 1245

Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro

1250 1255 1260

Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp

1265 1270 1275

Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser

1280 1285 1290

Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu

1295 1300 1305

Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser

1310 1315 1320

Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr

1325 1330 1335

Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser

1340 1345 1350

Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala

1355 1360 1365

Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr

1370 1375 1380

Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly

1385 1390 1395

Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His

1400 1405 1410

Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser

1415 1420 1425

Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser

1430 1435 1440

Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu

1445 1450 1455

Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala

1460 1465 1470

Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr

1475 1480 1485

Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile

1490 1495 1500

Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly

1505 1510 1515

Asp Ser Arg Ala Asp Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser

1520 1525 1530

Gly Ser Pro Lys Lys Lys Arg Lys Val Ser Pro Gly Ser Gly Ser

1535 1540 1545

Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Asn His

1550 1555 1560

Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro Ala

1565 1570 1575

Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile

1580 1585 1590

Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp

1595 1600 1605

Arg Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly

1610 1615 1620

Met Val Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg

1625 1630 1635

Ser Val Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu

1640 1645 1650

Val Ile Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro

1655 1660 1665

Ala Arg Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu

1670 1675 1680

Phe Tyr Arg Leu Leu His Asp Ala Arg Pro Lys Glu Gly Asp Asp

1685 1690 1695

Arg Pro Phe Phe Trp Leu Phe Glu Asn Val Val Ala Met Gly Val

1700 1705 1710

Ser Asp Lys Arg Asp Ile Ser Arg Phe Leu Glu Ser Asn Pro Val

1715 1720 1725

Met Ile Asp Ala Lys Glu Val Ser Ala Ala His Arg Ala Arg Tyr

1730 1735 1740

Phe Trp Gly Asn Leu Pro Gly Met Asn Arg Pro Leu Ala Ser Thr

1745 1750 1755

Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu Glu His Gly Arg

1760 1765 1770

Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr Arg Ser Asn

1775 1780 1785

Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe Met Asn

1790 1795 1800

Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val Phe

1805 1810 1815

Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu

1820 1825 1830

Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile

1835 1840 1845

Arg His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val Ser

1850 1855 1860

Ser Gly Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser

1865 1870 1875

Gly Leu Val Pro Leu Ser Leu Arg Gly Ser His Met Gly Pro Met

1880 1885 1890

Glu Ile Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln Pro Val Arg

1895 1900 1905

Val Leu Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys Ser Leu

1910 1915 1920

Gly Phe Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr Leu Lys

1925 1930 1935

Tyr Val Glu Asp Val Thr Asn Val Val Arg Arg Asp Val Glu Lys

1940 1945 1950

Trp Gly Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro Leu Gly

1955 1960 1965

Ser Ser Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln Phe His

1970 1975 1980

Arg Ile Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln Arg Pro

1985 1990 1995

Phe Phe Trp Ile Phe Met Asp Asn Leu Leu Leu Thr Glu Asp Asp

2000 2005 2010

Gln Glu Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala Val Thr Leu

2015 2020 2025

Gln Asp Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg Val Trp

2030 2035 2040

Ser Asn Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu Thr Pro

2045 2050 2055

Lys Glu Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg Ser Lys

2060 2065 2070

Leu Asp Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys Leu Leu

2075 2080 2085

Pro Leu Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser Leu Pro

2090 2095 2100

Leu Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Gly Ser Gly

2105 2110 2115

Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu

2120 2125 2130

Asn Pro Gly Pro Ser Glu Leu Ile Lys Glu Asn Met His Met Lys

2135 2140 2145

Leu Tyr Met Glu Gly Thr Val Asp Asn His His Phe Lys Cys Thr

2150 2155 2160

Ser Glu Gly Glu Gly Lys Pro Tyr Glu Gly Thr Gln Thr Met Arg

2165 2170 2175

Ile Lys Val Val Glu Gly Gly Pro Leu Pro Phe Ala Phe Asp Ile

2180 2185 2190

Leu Ala Thr Ser Phe Leu Tyr Gly Ser Lys Thr Phe Ile Asn His

2195 2200 2205

Thr Gln Gly Ile Pro Asp Phe Phe Lys Gln Ser Phe Pro Glu Gly

2210 2215 2220

Phe Thr Trp Glu Arg Val Thr Thr Tyr Glu Asp Gly Gly Val Leu

2225 2230 2235

Thr Ala Thr Gln Asp Thr Ser Leu Gln Asp Gly Cys Leu Ile Tyr

2240 2245 2250

Asn Val Lys Ile Arg Gly Val Asn Phe Thr Ser Asn Gly Pro Val

2255 2260 2265

Met Gln Lys Lys Thr Leu Gly Trp Glu Ala Phe Thr Glu Thr Leu

2270 2275 2280

Tyr Pro Ala Asp Gly Gly Leu Glu Gly Arg Asn Asp Met Ala Leu

2285 2290 2295

Lys Leu Val Gly Gly Ser His Leu Ile Ala Asn Ile Lys Thr Thr

2300 2305 2310

Tyr Arg Ser Lys Lys Pro Ala Lys Asn Leu Lys Met Pro Gly Val

2315 2320 2325

Tyr Tyr Val Asp Tyr Arg Leu Glu Arg Ile Lys Glu Ala Asn Asn

2330 2335 2340

Glu Thr Tyr Val Glu Gln His Glu Val Ala Val Ala Arg Tyr Cys

2345 2350 2355

Asp Leu Pro Ser Lys Leu Gly His Lys Leu Asn

2360 2365

<210> 14

<211> 2391

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 14

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro Gly Gly Pro Ser Ser Gly Ala Pro Pro

65 70 75 80

Pro Ser Gly Gly Ser Pro Ala Gly Ser Pro Thr Ser Thr Glu Glu Gly

85 90 95

Thr Ser Glu Ser Ala Thr Pro Glu Ser Gly Pro Gly Thr Ser Thr Glu

100 105 110

Pro Ser Glu Gly Ser Ala Pro Gly Ser Pro Ala Gly Ser Pro Thr Ser

115 120 125

Thr Glu Glu Gly Thr Ser Thr Glu Pro Ser Glu Gly Ser Ala Pro Gly

130 135 140

Thr Ser Thr Glu Pro Ser Glu Met Asp Lys Lys Tyr Ser Ile Gly Leu

145 150 155 160

Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr

165 170 175

Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His

180 185 190

Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu

195 200 205

Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr

210 215 220

Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu

225 230 235 240

Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe

245 250 255

Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn

260 265 270

Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His

275 280 285

Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu

290 295 300

Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu

305 310 315 320

Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe

325 330 335

Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile

340 345 350

Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser

355 360 365

Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys

370 375 380

Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr

385 390 395 400

Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln

405 410 415

Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln

420 425 430

Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser

435 440 445

Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr

450 455 460

Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His

465 470 475 480

Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu

485 490 495

Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly

500 505 510

Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys

515 520 525

Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu

530 535 540

Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser

545 550 555 560

Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg

565 570 575

Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu

580 585 590

Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg

595 600 605

Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile

610 615 620

Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln

625 630 635 640

Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu

645 650 655

Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr

660 665 670

Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro

675 680 685

Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe

690 695 700

Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe

705 710 715 720

Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp

725 730 735

Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile

740 745 750

Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu

755 760 765

Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu

770 775 780

Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys

785 790 795 800

Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys

805 810 815

Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp

820 825 830

Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile

835 840 845

His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val

850 855 860

Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly

865 870 875 880

Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp

885 890 895

Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile

900 905 910

Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser

915 920 925

Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser

930 935 940

Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu

945 950 955 960

Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp

965 970 975

Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp Ala Ile

980 985 990

Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu

995 1000 1005

Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser

1010 1015 1020

Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu

1025 1030 1035

Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys

1040 1045 1050

Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile

1055 1060 1065

Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala

1070 1075 1080

Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp

1085 1090 1095

Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu

1100 1105 1110

Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu

1115 1120 1125

Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val

1130 1135 1140

Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu

1145 1150 1155

Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile

1160 1165 1170

Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe

1175 1180 1185

Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu

1190 1195 1200

Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly

1205 1210 1215

Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr

1220 1225 1230

Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys

1235 1240 1245

Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro

1250 1255 1260

Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp

1265 1270 1275

Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser

1280 1285 1290

Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu

1295 1300 1305

Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser

1310 1315 1320

Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr

1325 1330 1335

Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser

1340 1345 1350

Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala

1355 1360 1365

Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr

1370 1375 1380

Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly

1385 1390 1395

Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His

1400 1405 1410

Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser

1415 1420 1425

Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser

1430 1435 1440

Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu

1445 1450 1455

Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala

1460 1465 1470

Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr

1475 1480 1485

Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile

1490 1495 1500

Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly

1505 1510 1515

Asp Ser Arg Ala Asp Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser

1520 1525 1530

Gly Ser Pro Lys Lys Lys Arg Lys Val Ser Pro Gly Ser Gly Ser

1535 1540 1545

Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Asn His

1550 1555 1560

Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro Ala

1565 1570 1575

Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile

1580 1585 1590

Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp

1595 1600 1605

Arg Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly

1610 1615 1620

Met Val Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg

1625 1630 1635

Ser Val Thr Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu

1640 1645 1650

Val Ile Gly Gly Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro

1655 1660 1665

Ala Arg Lys Gly Leu Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu

1670 1675 1680

Phe Tyr Arg Leu Leu His Asp Ala Arg Pro Lys Glu Gly Asp Asp

1685 1690 1695

Arg Pro Phe Phe Trp Leu Phe Glu Asn Val Val Ala Met Gly Val

1700 1705 1710

Ser Asp Lys Arg Asp Ile Ser Arg Phe Leu Glu Ser Asn Pro Val

1715 1720 1725

Met Ile Asp Ala Lys Glu Val Ser Ala Ala His Arg Ala Arg Tyr

1730 1735 1740

Phe Trp Gly Asn Leu Pro Gly Met Asn Arg Pro Leu Ala Ser Thr

1745 1750 1755

Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu Glu His Gly Arg

1760 1765 1770

Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr Arg Ser Asn

1775 1780 1785

Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe Met Asn

1790 1795 1800

Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val Phe

1805 1810 1815

Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu

1820 1825 1830

Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile

1835 1840 1845

Arg His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val Ser

1850 1855 1860

Ser Gly Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser

1865 1870 1875

Gly Leu Val Pro Leu Ser Leu Arg Gly Ser His Met Gly Pro Met

1880 1885 1890

Glu Ile Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln Pro Val Arg

1895 1900 1905

Val Leu Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys Ser Leu

1910 1915 1920

Gly Phe Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr Leu Lys

1925 1930 1935

Tyr Val Glu Asp Val Thr Asn Val Val Arg Arg Asp Val Glu Lys

1940 1945 1950

Trp Gly Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro Leu Gly

1955 1960 1965

Ser Ser Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln Phe His

1970 1975 1980

Arg Ile Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln Arg Pro

1985 1990 1995

Phe Phe Trp Ile Phe Met Asp Asn Leu Leu Leu Thr Glu Asp Asp

2000 2005 2010

Gln Glu Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala Val Thr Leu

2015 2020 2025

Gln Asp Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg Val Trp

2030 2035 2040

Ser Asn Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu Thr Pro

2045 2050 2055

Lys Glu Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg Ser Lys

2060 2065 2070

Leu Asp Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys Leu Leu

2075 2080 2085

Pro Leu Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser Leu Pro

2090 2095 2100

Leu Ser Arg Ala Asp Pro Lys Lys Lys Arg Lys Val Gly Ser Gly

2105 2110 2115

Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu

2120 2125 2130

Asn Pro Gly Pro Gly Ser Gly Ala Thr Asn Phe Ser Leu Leu Lys

2135 2140 2145

Gln Ala Gly Asp Val Glu Glu Asn Pro Gly Pro Ser Glu Leu Ile

2150 2155 2160

Lys Glu Asn Met His Met Lys Leu Tyr Met Glu Gly Thr Val Asp

2165 2170 2175

Asn His His Phe Lys Cys Thr Ser Glu Gly Glu Gly Lys Pro Tyr

2180 2185 2190

Glu Gly Thr Gln Thr Met Arg Ile Lys Val Val Glu Gly Gly Pro

2195 2200 2205

Leu Pro Phe Ala Phe Asp Ile Leu Ala Thr Ser Phe Leu Tyr Gly

2210 2215 2220

Ser Lys Thr Phe Ile Asn His Thr Gln Gly Ile Pro Asp Phe Phe

2225 2230 2235

Lys Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg Val Thr Thr

2240 2245 2250

Tyr Glu Asp Gly Gly Val Leu Thr Ala Thr Gln Asp Thr Ser Leu

2255 2260 2265

Gln Asp Gly Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly Val Asn

2270 2275 2280

Phe Thr Ser Asn Gly Pro Val Met Gln Lys Lys Thr Leu Gly Trp

2285 2290 2295

Glu Ala Phe Thr Glu Thr Leu Tyr Pro Ala Asp Gly Gly Leu Glu

2300 2305 2310

Gly Arg Asn Asp Met Ala Leu Lys Leu Val Gly Gly Ser His Leu

2315 2320 2325

Ile Ala Asn Ile Lys Thr Thr Tyr Arg Ser Lys Lys Pro Ala Lys

2330 2335 2340

Asn Leu Lys Met Pro Gly Val Tyr Tyr Val Asp Tyr Arg Leu Glu

2345 2350 2355

Arg Ile Lys Glu Ala Asn Asn Glu Thr Tyr Val Glu Gln His Glu

2360 2365 2370

Val Ala Val Ala Arg Tyr Cys Asp Leu Pro Ser Lys Leu Gly His

2375 2380 2385

Lys Leu Asn

2390

<210> 15

<211> 2345

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 15

Asn His Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro

1. 5 10 15

Ala Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile

20 25 30

Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp Arg

35 40 45

Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly Met Val

50 55 60

Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg Ser Val Thr

65 70 75 80

Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu Val Ile Gly Gly

85 90 95

Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro Ala Arg Lys Gly Leu

100 105 110

Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu Phe Tyr Arg Leu Leu His

115 120 125

Asp Ala Arg Pro Lys Glu Gly Asp Asp Arg Pro Phe Phe Trp Leu Phe

130 135 140

Glu Asn Val Val Ala Met Gly Val Ser Asp Lys Arg Asp Ile Ser Arg

145 150 155 160

Phe Leu Glu Ser Asn Pro Val Met Ile Asp Ala Lys Glu Val Ser Ala

165 170 175

Ala His Arg Ala Arg Tyr Phe Trp Gly Asn Leu Pro Gly Met Asn Arg

180 185 190

Pro Leu Ala Ser Thr Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu

195 200 205

Glu His Gly Arg Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr

210 215 220

Arg Ser Asn Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe

225 230 235 240

Met Asn Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val

245 250 255

Phe Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu

260 265 270

Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile Arg

275 280 285

His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val Ser Ser Gly

290 295 300

Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser Gly Leu Val

305 310 315 320

Pro Leu Ser Leu Arg Gly Ser His Met Gly Pro Met Glu Ile Tyr Lys

325 330 335

Thr Val Ser Ala Trp Lys Arg Gln Pro Val Arg Val Leu Ser Leu Phe

340 345 350

Arg Asn Ile Asp Lys Val Leu Lys Ser Leu Gly Phe Leu Glu Ser Gly

355 360 365

Ser Gly Ser Gly Gly Gly Thr Leu Lys Tyr Val Glu Asp Val Thr Asn

370 375 380

Val Val Arg Arg Asp Val Glu Lys Trp Gly Pro Phe Asp Leu Val Tyr

385 390 395 400

Gly Ser Thr Gln Pro Leu Gly Ser Ser Cys Asp Arg Cys Pro Gly Trp

405 410 415

Tyr Met Phe Gln Phe His Arg Ile Leu Gln Tyr Ala Leu Pro Arg Gln

420 425 430

Glu Ser Gln Arg Pro Phe Phe Trp Ile Phe Met Asp Asn Leu Leu Leu

435 440 445

Thr Glu Asp Asp Gln Glu Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala

450 455 460

Val Thr Leu Gln Asp Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg

465 470 475 480

Val Trp Ser Asn Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu Thr

485 490 495

Pro Lys Glu Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg Ser Lys

500 505 510

Leu Asp Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys Leu Leu Pro

515 520 525

Leu Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser Leu Pro Leu Gly

530 535 540

Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser Gly Gly Ser Pro Ala Gly

545 550 555 560

Ser Pro Thr Ser Thr Glu Glu Gly Thr Ser Glu Ser Ala Thr Pro Glu

565 570 575

Ser Gly Pro Gly Thr Ser Thr Glu Pro Ser Glu Gly Ser Ala Pro Gly

580 585 590

Ser Pro Ala Gly Ser Pro Thr Ser Thr Glu Glu Gly Thr Ser Thr Glu

595 600 605

Pro Ser Glu Gly Ser Ala Pro Gly Thr Ser Thr Glu Pro Ser Glu Met

610 615 620

Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly

625 630 635 640

Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys

645 650 655

Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly

660 665 670

Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys

675 680 685

Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr

690 695 700

Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe

705 710 715 720

Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His

725 730 735

Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His

740 745 750

Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser

755 760 765

Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met

770 775 780

Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp

785 790 795 800

Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn

805 810 815

Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys

820 825 830

Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu

835 840 845

Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu

850 855 860

Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp

865 870 875 880

Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp

885 890 895

Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu

900 905 910

Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile

915 920 925

Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met

930 935 940

Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala

945 950 955 960

Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp

965 970 975

Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln

980 985 990

Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly

995 1000 1005

Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg

1010 1015 1020

Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His

1025 1030 1035

Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr

1040 1045 1050

Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr

1055 1060 1065

Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser

1070 1075 1080

Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro

1085 1090 1095

Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser

1100 1105 1110

Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu

1115 1120 1125

Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val

1130 1135 1140

Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg

1145 1150 1155

Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp

1160 1165 1170

Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys

1175 1180 1185

Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile

1190 1195 1200

Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His

1205 1210 1215

Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu

1220 1225 1230

Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu

1235 1240 1245

Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala

1250 1255 1260

His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg

1265 1270 1275

Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile

1280 1285 1290

Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser

1295 1300 1305

Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp

1310 1315 1320

Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly

1325 1330 1335

Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser

1340 1345 1350

Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp

1355 1360 1365

Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val

1370 1375 1380

Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys

1385 1390 1395

Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu

1400 1405 1410

Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln

1415 1420 1425

Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg

1430 1435 1440

Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp

1445 1450 1455

Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys Asp Asp

1460 1465 1470

Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly

1475 1480 1485

Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys

1490 1495 1500

Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg

1505 1510 1515

Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu

1520 1525 1530

Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg

1535 1540 1545

Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn

1550 1555 1560

Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val

1565 1570 1575

Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe

1580 1585 1590

Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr His His Ala His

1595 1600 1605

Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys

1610 1615 1620

Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val

1625 1630 1635

Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu Gln Glu Ile Gly

1640 1645 1650

Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe

1655 1660 1665

Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg

1670 1675 1680

Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile Val Trp Asp

1685 1690 1695

Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser Met Pro

1700 1705 1710

Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe

1715 1720 1725

Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile

1730 1735 1740

Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp

1745 1750 1755

Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val Ala Lys Val Glu

1760 1765 1770

Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly

1775 1780 1785

Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp

1790 1795 1800

Phe Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile

1805 1810 1815

Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg

1820 1825 1830

Lys Arg Met Leu Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu

1835 1840 1845

Leu Ala Leu Pro Ser Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser

1850 1855 1860

His Tyr Glu Lys Leu Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys

1865 1870 1875

Gln Leu Phe Val Glu Gln His Lys His Tyr Leu Asp Glu Ile Ile

1880 1885 1890

Glu Gln Ile Ser Glu Phe Ser Lys Arg Val Ile Leu Ala Asp Ala

1895 1900 1905

Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys His Arg Asp Lys

1910 1915 1920

Pro Ile Arg Glu Gln Ala Glu Asn Ile Ile His Leu Phe Thr Leu

1925 1930 1935

Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr

1940 1945 1950

Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala

1955 1960 1965

Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile

1970 1975 1980

Asp Leu Ser Gln Leu Gly Gly Asp Ala Tyr Pro Tyr Asp Val Pro

1985 1990 1995

Asp Tyr Ala Ser Leu Gly Ser Gly Ser Pro Lys Lys Lys Arg Lys

2000 2005 2010

Val Glu Asp Pro Lys Lys Lys Arg Lys Val Asp Gly Ile Gly Ser

2015 2020 2025

Gly Ser Asn Gly Ser Ser Gly Ser Ser Glu Leu Ile Lys Glu Asn

2030 2035 2040

Met His Met Lys Leu Tyr Met Glu Gly Thr Val Asp Asn His His

2045 2050 2055

Phe Lys Cys Thr Ser Glu Gly Glu Gly Lys Pro Tyr Glu Gly Thr

2060 2065 2070

Gln Thr Met Arg Ile Lys Val Val Glu Gly Gly Pro Leu Pro Phe

2075 2080 2085

Ala Phe Asp Ile Leu Ala Thr Ser Phe Leu Tyr Gly Ser Lys Thr

2090 2095 2100

Phe Ile Asn His Thr Gln Gly Ile Pro Asp Phe Phe Lys Gln Ser

2105 2110 2115

Phe Pro Glu Gly Phe Thr Trp Glu Arg Val Thr Thr Tyr Glu Asp

2120 2125 2130

Gly Gly Val Leu Thr Ala Thr Gln Asp Thr Ser Leu Gln Asp Gly

2135 2140 2145

Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly Val Asn Phe Thr Ser

2150 2155 2160

Asn Gly Pro Val Met Gln Lys Lys Thr Leu Gly Trp Glu Ala Phe

2165 2170 2175

Thr Glu Thr Leu Tyr Pro Ala Asp Gly Gly Leu Glu Gly Arg Asn

2180 2185 2190

Asp Met Ala Leu Lys Leu Val Gly Gly Ser His Leu Ile Ala Asn

2195 2200 2205

Ile Lys Thr Thr Tyr Arg Ser Lys Lys Pro Ala Lys Asn Leu Lys

2210 2215 2220

Met Pro Gly Val Tyr Tyr Val Asp Tyr Arg Leu Glu Arg Ile Lys

2225 2230 2235

Glu Ala Asn Asn Glu Thr Tyr Val Glu Gln His Glu Val Ala Val

2240 2245 2250

Ala Arg Tyr Cys Asp Leu Pro Ser Lys Leu Gly His Lys Leu Asn

2255 2260 2265

Gly Gly Gly Gly Gly Met Asp Ala Lys Ser Leu Thr Ala Trp Ser

2270 2275 2280

Arg Thr Leu Val Thr Phe Lys Asp Val Phe Val Asp Phe Thr Arg

2285 2290 2295

Glu Glu Trp Lys Leu Leu Asp Thr Ala Gln Gln Ile Val Tyr Arg

2300 2305 2310

Asn Val Met Leu Glu Asn Tyr Lys Asn Leu Val Ser Leu Gly Tyr

2315 2320 2325

Gln Leu Thr Lys Pro Asp Val Ile Leu Arg Leu Glu Lys Gly Glu

2330 2335 2340

Glu Pro

2345

<210> 16

<211> 71

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 16

Asp Ala Lys Ser Leu Thr Ala Trp Ser Arg Thr Leu Val Thr Phe Lys

1. 5 10 15

Asp Val Phe Val Asp Phe Thr Arg Glu Glu Trp Lys Leu Leu Asp Thr

20 25 30

Ala Gln Gln Ile Val Tyr Arg Asn Val Met Leu Glu Asn Tyr Lys Asn

35 40 45

Leu Val Ser Leu Gly Tyr Gln Leu Thr Lys Pro Asp Val Ile Leu Arg

50 55 60

Leu Glu Lys Gly Glu Glu Pro

65 70

<210> 17

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 17

Gly Gly Ser Gly Gly Gly Ser

15

<210> 18

<211> 3

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 18

Ser Gly Ser

1

<210> 19

<211> 17

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 19

Glu Ala Ser Gly Ser Gly Arg Ala Ser Pro Gly Ile Pro Gly Ser Thr

1. 5 10 15

Arg

<210> 20

<211> 4

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 20

Ser Arg Ala Asp

1

<210> 21

<211> 3

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 21

Gly Ser Gly

1

<210> 22

<211> 3

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 22

Ser Pro Gly

1

<210> 23

<211> 1368

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 23

Met Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val

1. 5 10 15

Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe

20 25 30

Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile

35 40 45

Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu

50 55 60

Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys

65 70 75 80

Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser

85 90 95

Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys

100 105 110

His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr

115 120 125

His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp

130 135 140

Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His

145 150 155 160

Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro

165 170 175

Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr

180 185 190

Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala

195 200 205

Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn

210 215 220

Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn

225 230 235 240

Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe

245 250 255

Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp

260 265 270

Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp

275 280 285

Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp

290 295 300

Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser

305 310 315 320

Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys

325 330 335

Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe

340 345 350

Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser

355 360 365

Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp

370 375 380

Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg

385 390 395 400

Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu

405 410 415

Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe

420 425 430

Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile

435 440 445

Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp

450 455 460

Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu

465 470 475 480

Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr

485 490 495

Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser

500 505 510

Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys

515 520 525

Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln

530 535 540

Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr

545 550 555 560

Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp

565 570 575

Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly

580 585 590

Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp

595 600 605

Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr

610 615 620

Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala

625 630 635 640

His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr

645 650 655

Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp

660 665 670

Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe

675 680 685

Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe

690 695 700

Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu

705 710 715 720

His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly

725 730 735

Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly

740 745 750

Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln

755 760 765

Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile

770 775 780

Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro

785 790 795 800

Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu

805 810 815

Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg

820 825 830

Leu Ser Asp Tyr Asp Val Asp Ala Ile Val Pro Gln Ser Phe Leu Lys

835 840 845

Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg

850 855 860

Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys

865 870 875 880

Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys

885 890 895

Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp

900 905 910

Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr

915 920 925

Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp

930 935 940

Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser

945 950 955 960

Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg

965 970 975

Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val

980 985 990

Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe

995 1000 1005

Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala

1010 1015 1020

Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe

1025 1030 1035

Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala

1040 1045 1050

Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu

1055 1060 1065

Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val

1070 1075 1080

Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr

1085 1090 1095

Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys

1100 1105 1110

Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro

1115 1120 1125

Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val

1130 1135 1140

Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys

1145 1150 1155

Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser

1160 1165 1170

Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys

1175 1180 1185

Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu

1190 1195 1200

Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly

1205 1210 1215

Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val

1220 1225 1230

Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser

1235 1240 1245

Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys

1250 1255 1260

His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys

1265 1270 1275

Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala

1280 1285 1290

Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn

1295 1300 1305

Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala

1310 1315 1320

Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser

1325 1330 1335

Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr

1340 1345 1350

Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp

1355 1360 1365

<210> 24

<211> 9

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 24

Tyr Pro Tyr Asp Val Pro Asp Tyr Ala

15

<210> 25

<211> 7

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 25

Pro Lys Lys Lys Arg Lys Val

15

<210> 26

<211> 301

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 26

Asn His Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro

1. 5 10 15

Ala Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile

20 25 30

Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp Arg

35 40 45

Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly Met Val

50 55 60

Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg Ser Val Thr

65 70 75 80

Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu Val Ile Gly Gly

85 90 95

Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro Ala Arg Lys Gly Leu

100 105 110

Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu Phe Tyr Arg Leu Leu His

115 120 125

Asp Ala Arg Pro Lys Glu Gly Asp Asp Arg Pro Phe Phe Trp Leu Phe

130 135 140

Glu Asn Val Val Ala Met Gly Val Ser Asp Lys Arg Asp Ile Ser Arg

145 150 155 160

Phe Leu Glu Ser Asn Pro Val Met Ile Asp Ala Lys Glu Val Ser Ala

165 170 175

Ala His Arg Ala Arg Tyr Phe Trp Gly Asn Leu Pro Gly Met Asn Arg

180 185 190

Pro Leu Ala Ser Thr Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu

195 200 205

Glu His Gly Arg Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr

210 215 220

Arg Ser Asn Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe

225 230 235 240

Met Asn Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val

245 250 255

Phe Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu

260 265 270

Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile Arg

275 280 285

His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val

290 295 300

<210> 27

<211> 27

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 27

Ser Ser Gly Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser

1. 5 10 15

Gly Leu Val Pro Leu Ser Leu Arg Gly Ser His

20 25

<210> 28

<211> 215

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 28

Met Gly Pro Met Glu Ile Tyr Lys Thr Val Ser Ala Trp Lys Arg Gln

1. 5 10 15

Pro Val Arg Val Leu Ser Leu Phe Arg Asn Ile Asp Lys Val Leu Lys

20 25 30

Ser Leu Gly Phe Leu Glu Ser Gly Ser Gly Ser Gly Gly Gly Thr Leu

35 40 45

Lys Tyr Val Glu Asp Val Thr Asn Val Val Arg Arg Asp Val Glu Lys

50 55 60

Trp Gly Pro Phe Asp Leu Val Tyr Gly Ser Thr Gln Pro Leu Gly Ser

65 70 75 80

Ser Cys Asp Arg Cys Pro Gly Trp Tyr Met Phe Gln Phe His Arg Ile

85 90 95

Leu Gln Tyr Ala Leu Pro Arg Gln Glu Ser Gln Arg Pro Phe Phe Trp

100 105 110

Ile Phe Met Asp Asn Leu Leu Leu Thr Glu Asp Asp Gln Glu Thr Thr

115 120 125

Thr Arg Phe Leu Gln Thr Glu Ala Val Thr Leu Gln Asp Val Arg Gly

130 135 140

Arg Asp Tyr Gln Asn Ala Met Arg Val Trp Ser Asn Ile Pro Gly Leu

145 150 155 160

Lys Ser Lys His Ala Pro Leu Thr Pro Lys Glu Glu Glu Tyr Leu Gln

165 170 175

Ala Gln Val Arg Ser Arg Ser Lys Leu Asp Ala Pro Lys Val Asp Leu

180 185 190

Leu Val Lys Asn Cys Leu Leu Pro Leu Arg Glu Tyr Phe Lys Tyr Phe

195 200 205

Ser Gln Asn Ser Leu Pro Leu

210 215

<210> 29

<211> 19

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 29

Ala Thr Asn Phe Ser Leu Leu Lys Gln Ala Gly Asp Val Glu Glu Asn

1. 5 10 15

Pro Gly Pro

<210> 30

<211> 232

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 30

Ser Glu Leu Ile Lys Glu Asn Met His Met Lys Leu Tyr Met Glu Gly

1. 5 10 15

Thr Val Asp Asn His His Phe Lys Cys Thr Ser Glu Gly Glu Gly Lys

20 25 30

Pro Tyr Glu Gly Thr Gln Thr Met Arg Ile Lys Val Val Glu Gly Gly

35 40 45

Pro Leu Pro Phe Ala Phe Asp Ile Leu Ala Thr Ser Phe Leu Tyr Gly

50 55 60

Ser Lys Thr Phe Ile Asn His Thr Gln Gly Ile Pro Asp Phe Phe Lys

65 70 75 80

Gln Ser Phe Pro Glu Gly Phe Thr Trp Glu Arg Val Thr Thr Tyr Glu

85 90 95

Asp Gly Gly Val Leu Thr Ala Thr Gln Asp Thr Ser Leu Gln Asp Gly

100 105 110

Cys Leu Ile Tyr Asn Val Lys Ile Arg Gly Val Asn Phe Thr Ser Asn

115 120 125

Gly Pro Val Met Gln Lys Lys Thr Leu Gly Trp Glu Ala Phe Thr Glu

130 135 140

Thr Leu Tyr Pro Ala Asp Gly Gly Leu Glu Gly Arg Asn Asp Met Ala

145 150 155 160

Leu Lys Leu Val Gly Gly Ser His Leu Ile Ala Asn Ile Lys Thr Thr

165 170 175

Tyr Arg Ser Lys Lys Pro Ala Lys Asn Leu Lys Met Pro Gly Val Tyr

180 185 190

Tyr Val Asp Tyr Arg Leu Glu Arg Ile Lys Glu Ala Asn Asn Glu Thr

195 200 205

Tyr Val Glu Gln His Glu Val Ala Val Ala Arg Tyr Cys Asp Leu Pro

210 215 220

Ser Lys Leu Gly His Lys Leu Asn

225 230

<210> 31

<211> 16

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 31

Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser

1. 5 10 15

<210> 32

<211> 80

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 32

Gly Gly Pro Ser Ser Gly Ala Pro Pro Pro Ser Gly Gly Ser Pro Ala

1. 5 10 15

Gly Ser Pro Thr Ser Thr Glu Glu Gly Thr Ser Glu Ser Ala Thr Pro

20 25 30

Glu Ser Gly Pro Gly Thr Ser Thr Glu Pro Ser Glu Gly Ser Ala Pro

35 40 45

Gly Ser Pro Ala Gly Ser Pro Thr Ser Thr Glu Glu Gly Thr Ser Thr

50 55 60

Glu Pro Ser Glu Gly Ser Ala Pro Gly Thr Ser Thr Glu Pro Ser Glu

65 70 75 80

<210> 33

<211> 543

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 33

Asn His Asp Gln Glu Phe Asp Pro Pro Lys Val Tyr Pro Pro Val Pro

1. 5 10 15

Ala Glu Lys Arg Lys Pro Ile Arg Val Leu Ser Leu Phe Asp Gly Ile

20 25 30

Ala Thr Gly Leu Leu Val Leu Lys Asp Leu Gly Ile Gln Val Asp Arg

35 40 45

Tyr Ile Ala Ser Glu Val Cys Glu Asp Ser Ile Thr Val Gly Met Val

50 55 60

Arg His Gln Gly Lys Ile Met Tyr Val Gly Asp Val Arg Ser Val Thr

65 70 75 80

Gln Lys His Ile Gln Glu Trp Gly Pro Phe Asp Leu Val Ile Gly Gly

85 90 95

Ser Pro Cys Asn Asp Leu Ser Ile Val Asn Pro Ala Arg Lys Gly Leu

100 105 110

Tyr Glu Gly Thr Gly Arg Leu Phe Phe Glu Phe Tyr Arg Leu Leu His

115 120 125

Asp Ala Arg Pro Lys Glu Gly Asp Asp Arg Pro Phe Phe Trp Leu Phe

130 135 140

Glu Asn Val Val Ala Met Gly Val Ser Asp Lys Arg Asp Ile Ser Arg

145 150 155 160

Phe Leu Glu Ser Asn Pro Val Met Ile Asp Ala Lys Glu Val Ser Ala

165 170 175

Ala His Arg Ala Arg Tyr Phe Trp Gly Asn Leu Pro Gly Met Asn Arg

180 185 190

Pro Leu Ala Ser Thr Val Asn Asp Lys Leu Glu Leu Gln Glu Cys Leu

195 200 205

Glu His Gly Arg Ile Ala Lys Phe Ser Lys Val Arg Thr Ile Thr Thr

210 215 220

Arg Ser Asn Ser Ile Lys Gln Gly Lys Asp Gln His Phe Pro Val Phe

225 230 235 240

Met Asn Glu Lys Glu Asp Ile Leu Trp Cys Thr Glu Met Glu Arg Val

245 250 255

Phe Gly Phe Pro Val His Tyr Thr Asp Val Ser Asn Met Ser Arg Leu

260 265 270

Ala Arg Gln Arg Leu Leu Gly Arg Ser Trp Ser Val Pro Val Ile Arg

275 280 285

His Leu Phe Ala Pro Leu Lys Glu Tyr Phe Ala Cys Val Ser Ser Gly

290 295 300

Asn Ser Asn Ala Asn Ser Arg Gly Pro Ser Phe Ser Ser Gly Leu Val

305 310 315 320

Pro Leu Ser Leu Arg Gly Ser His Met Gly Pro Met Glu Ile Tyr Lys

325 330 335

Thr Val Ser Ala Trp Lys Arg Gln Pro Val Arg Val Leu Ser Leu Phe

340 345 350

Arg Asn Ile Asp Lys Val Leu Lys Ser Leu Gly Phe Leu Glu Ser Gly

355 360 365

Ser Gly Ser Gly Gly Gly Thr Leu Lys Tyr Val Glu Asp Val Thr Asn

370 375 380

Val Val Arg Arg Asp Val Glu Lys Trp Gly Pro Phe Asp Leu Val Tyr

385 390 395 400

Gly Ser Thr Gln Pro Leu Gly Ser Ser Cys Asp Arg Cys Pro Gly Trp

405 410 415

Tyr Met Phe Gln Phe His Arg Ile Leu Gln Tyr Ala Leu Pro Arg Gln

420 425 430

Glu Ser Gln Arg Pro Phe Phe Trp Ile Phe Met Asp Asn Leu Leu Leu

435 440 445

Thr Glu Asp Asp Gln Glu Thr Thr Thr Arg Phe Leu Gln Thr Glu Ala

450 455 460

Val Thr Leu Gln Asp Val Arg Gly Arg Asp Tyr Gln Asn Ala Met Arg

465 470 475 480

Val Trp Ser Asn Ile Pro Gly Leu Lys Ser Lys His Ala Pro Leu Thr

485 490 495

Pro Lys Glu Glu Glu Tyr Leu Gln Ala Gln Val Arg Ser Arg Ser Lys

500 505 510

Leu Asp Ala Pro Lys Val Asp Leu Leu Val Lys Asn Cys Leu Leu Pro

515 520 525

Leu Arg Glu Tyr Phe Lys Tyr Phe Ser Gln Asn Ser Leu Pro Leu

530 535 540

<210> 34

<211> 1307

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 34

Met Thr Gln Phe Glu Gly Phe Thr Asn Leu Tyr Gln Val Ser Lys Thr

1. 5 10 15

Leu Arg Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Lys His Ile Gln

20 25 30

Glu Gln Gly Phe Ile Glu Glu Asp Lys Ala Arg Asn Asp His Tyr Lys

35 40 45

Glu Leu Lys Pro Ile Ile Asp Arg Ile Tyr Lys Thr Tyr Ala Asp Gln

50 55 60

Cys Leu Gln Leu Val Gln Leu Asp Trp Glu Asn Leu Ser Ala Ala Ile

65 70 75 80

Asp Ser Tyr Arg Lys Glu Lys Thr Glu Glu Thr Arg Asn Ala Leu Ile

85 90 95

Glu Glu Gln Ala Thr Tyr Arg Asn Ala Ile His Asp Tyr Phe Ile Gly

100 105 110

Arg Thr Asp Asn Leu Thr Asp Ala Ile Asn Lys Arg His Ala Glu Ile

115 120 125

Tyr Lys Gly Leu Phe Lys Ala Glu Leu Phe Asn Gly Lys Val Leu Lys

130 135 140

Gln Leu Gly Thr Val Thr Thr Thr Glu His Glu Asn Ala Leu Leu Arg

145 150 155 160

Ser Phe Asp Lys Phe Thr Thr Tyr Phe Ser Gly Phe Tyr Glu Asn Arg

165 170 175

Lys Asn Val Phe Ser Ala Glu Asp Ile Ser Thr Ala Ile Pro His Arg

180 185 190

Ile Val Gln Asp Asn Phe Pro Lys Phe Lys Glu Asn Cys His Ile Phe

195 200 205

Thr Arg Leu Ile Thr Ala Val Pro Ser Leu Arg Glu His Phe Glu Asn

210 215 220

Val Lys Lys Ala Ile Gly Ile Phe Val Ser Thr Ser Ile Glu Glu Val

225 230 235 240

Phe Ser Phe Pro Phe Tyr Asn Gln Leu Leu Thr Gln Thr Gln Ile Asp

245 250 255

Leu Tyr Asn Gln Leu Leu Gly Gly Ile Ser Arg Glu Ala Gly Thr Glu

260 265 270

Lys Ile Lys Gly Leu Asn Glu Val Leu Asn Leu Ala Ile Gln Lys Asn

275 280 285

Asp Glu Thr Ala His Ile Ile Ala Ser Leu Pro His Arg Phe Ile Pro

290 295 300

Leu Phe Lys Gln Ile Leu Ser Asp Arg Asn Thr Leu Ser Phe Ile Leu

305 310 315 320

Glu Glu Phe Lys Ser Asp Glu Glu Val Ile Gln Ser Phe Cys Lys Tyr

325 330 335

Lys Thr Leu Leu Arg Asn Glu Asn Val Leu Glu Thr Ala Glu Ala Leu

340 345 350

Phe Asn Glu Leu Asn Ser Ile Asp Leu Thr His Ile Phe Ile Ser His

355 360 365

Lys Lys Leu Glu Thr Ile Ser Ser Ala Leu Cys Asp His Trp Asp Thr

370 375 380

Leu Arg Asn Ala Leu Tyr Glu Arg Arg Ile Ser Glu Leu Thr Gly Lys

385 390 395 400

Ile Thr Lys Ser Ala Lys Glu Lys Val Gln Arg Ser Leu Lys His Glu

405 410 415

Asp Ile Asn Leu Gln Glu Ile Ile Ser Ala Ala Gly Lys Glu Leu Ser

420 425 430

Glu Ala Phe Lys Gln Lys Thr Ser Glu Ile Leu Ser His Ala His Ala

435 440 445

Ala Leu Asp Gln Pro Leu Pro Thr Thr Leu Lys Lys Gln Glu Glu Lys

450 455 460

Glu Ile Leu Lys Ser Gln Leu Asp Ser Leu Leu Gly Leu Tyr His Leu

465 470 475 480

Leu Asp Trp Phe Ala Val Asp Glu Ser Asn Glu Val Asp Pro Glu Phe

485 490 495

Ser Ala Arg Leu Thr Gly Ile Lys Leu Glu Met Glu Pro Ser Leu Ser

500 505 510

Phe Tyr Asn Lys Ala Arg Asn Tyr Ala Thr Lys Lys Pro Tyr Ser Val

515 520 525

Glu Lys Phe Lys Leu Asn Phe Gln Met Pro Thr Leu Ala Ser Gly Trp

530 535 540

Asp Val Asn Lys Glu Lys Asn Asn Gly Ala Ile Leu Phe Val Lys Asn

545 550 555 560

Gly Leu Tyr Tyr Leu Gly Ile Met Pro Lys Gln Lys Gly Arg Tyr Lys

565 570 575

Ala Leu Ser Phe Glu Pro Thr Glu Lys Thr Ser Glu Gly Phe Asp Lys

580 585 590

Met Tyr Tyr Asp Tyr Phe Pro Asp Ala Ala Lys Met Ile Pro Lys Cys

595 600 605

Ser Thr Gln Leu Lys Ala Val Thr Ala His Phe Gln Thr His Thr Thr

610 615 620

Pro Ile Leu Leu Ser Asn Asn Phe Ile Glu Pro Leu Glu Ile Thr Lys

625 630 635 640

Glu Ile Tyr Asp Leu Asn Asn Pro Glu Lys Glu Pro Lys Lys Phe Gln

645 650 655

Thr Ala Tyr Ala Lys Lys Thr Gly Asp Gln Lys Gly Tyr Arg Glu Ala

660 665 670

Leu Cys Lys Trp Ile Asp Phe Thr Arg Asp Phe Leu Ser Lys Tyr Thr

675 680 685

Lys Thr Thr Ser Ile Asp Leu Ser Ser Leu Arg Pro Ser Ser Gln Tyr

690 695 700

Lys Asp Leu Gly Glu Tyr Tyr Ala Glu Leu Asn Pro Leu Leu Tyr His

705 710 715 720

Ile Ser Phe Gln Arg Ile Ala Glu Lys Glu Ile Met Asp Ala Val Glu

725 730 735

Thr Gly Lys Leu Tyr Leu Phe Gln Ile Tyr Asn Lys Asp Phe Ala Lys

740 745 750

Gly His His Gly Lys Pro Asn Leu His Thr Leu Tyr Trp Thr Gly Leu

755 760 765

Phe Ser Pro Glu Asn Leu Ala Lys Thr Ser Ile Lys Leu Asn Gly Gln

770 775 780

Ala Glu Leu Phe Tyr Arg Pro Lys Ser Arg Met Lys Arg Met Ala His

785 790 795 800

Arg Leu Gly Glu Lys Met Leu Asn Lys Lys Leu Lys Asp Gln Lys Thr

805 810 815

Pro Ile Pro Asp Thr Leu Tyr Gln Glu Leu Tyr Asp Tyr Val Asn His

820 825 830

Arg Leu Ser His Asp Leu Ser Asp Glu Ala Arg Ala Leu Leu Pro Asn

835 840 845

Val Ile Thr Lys Glu Val Ser His Glu Ile Ile Lys Asp Arg Arg Phe

850 855 860

Thr Ser Asp Lys Phe Phe Phe His Val Pro Ile Thr Leu Asn Tyr Gln

865 870 875 880

Ala Ala Asn Ser Pro Ser Lys Phe Asn Gln Arg Val Asn Ala Tyr Leu

885 890 895

Lys Glu His Pro Glu Thr Pro Ile Ile Gly Ile Asp Arg Gly Glu Arg

900 905 910

Asn Leu Ile Tyr Ile Thr Val Ile Asp Ser Thr Gly Lys Ile Leu Glu

915 920 925

Gln Arg Ser Leu Asn Thr Ile Gln Gln Phe Asp Tyr Gln Lys Lys Leu

930 935 940

Asp Asn Arg Glu Lys Glu Arg Val Ala Ala Arg Gln Ala Trp Ser Val

945 950 955 960

Val Gly Thr Ile Lys Asp Leu Lys Gln Gly Tyr Leu Ser Gln Val Ile

965 970 975

His Glu Ile Val Asp Leu Met Ile His Tyr Gln Ala Val Val Val Leu

980 985 990

Ala Asn Leu Asn Phe Gly Phe Lys Ser Lys Arg Thr Gly Ile Ala Glu

995 1000 1005

Lys Ala Val Tyr Gln Gln Phe Glu Lys Met Leu Ile Asp Lys Leu

1010 1015 1020

Asn Cys Leu Val Leu Lys Asp Tyr Pro Ala Glu Lys Val Gly Gly

1025 1030 1035

Val Leu Asn Pro Tyr Gln Leu Thr Asp Gln Phe Thr Ser Phe Ala

1040 1045 1050

Lys Met Gly Thr Gln Ser Gly Phe Leu Phe Tyr Val Pro Ala Pro

1055 1060 1065

Tyr Thr Ser Lys Ile Asp Pro Leu Thr Gly Phe Val Asp Pro Phe

1070 1075 1080

Val Trp Lys Thr Ile Lys Asn His Glu Ser Arg Lys His Phe Leu

1085 1090 1095

Glu Gly Phe Asp Phe Leu His Tyr Asp Val Lys Thr Gly Asp Phe

1100 1105 1110

Ile Leu His Phe Lys Met Asn Arg Asn Leu Ser Phe Gln Arg Gly

1115 1120 1125

Leu Pro Gly Phe Met Pro Ala Trp Asp Ile Val Phe Glu Lys Asn

1130 1135 1140

Glu Thr Gln Phe Asp Ala Lys Gly Thr Pro Phe Ile Ala Gly Lys

1145 1150 1155

Arg Ile Val Pro Val Ile Glu Asn His Arg Phe Thr Gly Arg Tyr

1160 1165 1170

Arg Asp Leu Tyr Pro Ala Asn Glu Leu Ile Ala Leu Leu Glu Glu

1175 1180 1185

Lys Gly Ile Val Phe Arg Asp Gly Ser Asn Ile Leu Pro Lys Leu

1190 1195 1200

Leu Glu Asn Asp Asp Ser His Ala Ile Asp Thr Met Val Ala Leu

1205 1210 1215

Ile Arg Ser Val Leu Gln Met Arg Asn Ser Asn Ala Ala Thr Gly

1220 1225 1230

Glu Asp Tyr Ile Asn Ser Pro Val Arg Asp Leu Asn Gly Val Cys

1235 1240 1245

Phe Asp Ser Arg Phe Gln Asn Pro Glu Trp Pro Met Asp Ala Asp

1250 1255 1260

Ala Asn Gly Ala Tyr His Ile Ala Leu Lys Gly Gln Leu Leu Leu

1265 1270 1275

Asn His Leu Lys Glu Ser Lys Asp Leu Lys Leu Gln Asn Gly Ile

1280 1285 1290

Ser Asn Gln Asp Trp Leu Ala Tyr Ile Gln Glu Leu Arg Asn

1295 1300 1305

<210> 35

<211> 1228

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 35

Met Ser Lys Leu Glu Lys Phe Thr Asn Cys Tyr Ser Leu Ser Lys Thr

1.5 10 15

Leu Arg Phe Lys Ala Ile Pro Val Gly Lys Thr Gln Glu Asn Ile Asp

20 25 30

Asn Lys Arg Leu Leu Val Glu Asp Glu Lys Arg Ala Glu Asp Tyr Lys

35 40 45

Gly Val Lys Lys Leu Leu Asp Arg Tyr Tyr Leu Ser Phe Ile Asn Asp

50 55 60

Val Leu His Ser Ile Lys Leu Lys Asn Leu Asn Asn Tyr Ile Ser Leu

65 70 75 80

Phe Arg Lys Lys Thr Arg Thr Glu Lys Glu Asn Lys Glu Leu Glu Asn

85 90 95

Leu Glu Ile Asn Leu Arg Lys Glu Ile Ala Lys Ala Phe Lys Gly Asn

100 105 110

Glu Gly Tyr Lys Ser Leu Phe Lys Lys Asp Ile Ile Glu Thr Ile Leu

115 120 125

Pro Glu Phe Leu Asp Asp Lys Asp Glu Ile Ala Leu Val Asn Ser Phe

130 135 140

Asn Gly Phe Thr Thr Ala Phe Thr Gly Phe Phe Asp Asn Arg Glu Asn

145 150 155 160

Met Phe Ser Glu Glu Ala Lys Ser Thr Ser Ile Ala Phe Arg Cys Ile

165 170 175

Asn Glu Asn Leu Thr Arg Tyr Ile Ser Asn Met Asp Ile Phe Glu Lys

180 185 190

Val Asp Ala Ile Phe Asp Lys His Glu Val Gln Glu Ile Lys Glu Lys

195 200 205

Ile Leu Asn Ser Asp Tyr Asp Val Glu Asp Phe Phe Glu Gly Glu Phe

210 215 220

Phe Asn Phe Val Leu Thr Gln Glu Gly Ile Asp Val Tyr Asn Ala Ile

225 230 235 240

Ile Gly Gly Phe Val Thr Glu Ser Gly Glu Lys Ile Lys Gly Leu Asn

245 250 255

Glu Tyr Ile Asn Leu Tyr Asn Gln Lys Thr Lys Gln Lys Leu Pro Lys

260 265 270

Phe Lys Pro Leu Tyr Lys Gln Val Leu Ser Asp Arg Glu Ser Leu Ser

275 280 285

Phe Tyr Gly Glu Gly Tyr Thr Ser Asp Glu Glu Val Leu Glu Val Phe

290 295 300

Arg Asn Thr Leu Asn Lys Asn Ser Glu Ile Phe Ser Ser Ile Lys Lys

305 310 315 320

Leu Glu Lys Leu Phe Lys Asn Phe Asp Glu Tyr Ser Ser Ala Gly Ile

325 330 335

Phe Val Lys Asn Gly Pro Ala Ile Ser Thr Ile Ser Lys Asp Ile Phe

340 345 350

Gly Glu Trp Asn Val Ile Arg Asp Lys Trp Asn Ala Glu Tyr Asp Asp

355 360 365

Ile His Leu Lys Lys Lys Ala Val Val Thr Glu Lys Tyr Glu Asp Asp

370 375 380

Arg Arg Lys Ser Phe Lys Lys Ile Gly Ser Phe Ser Leu Glu Gln Leu

385 390 395 400

Gln Glu Tyr Ala Asp Ala Asp Leu Ser Val Val Glu Lys Leu Lys Glu

405 410 415

Ile Ile Ile Gln Lys Val Asp Glu Ile Tyr Lys Val Tyr Gly Ser Ser

420 425 430

Glu Lys Leu Phe Asp Ala Asp Phe Val Leu Glu Lys Ser Leu Lys Lys

435 440 445

Asn Asp Ala Val Val Ala Ile Met Lys Asp Leu Leu Asp Ser Val Lys

450 455 460

Ser Phe Glu Asn Tyr Ile Lys Ala Phe Phe Gly Glu Gly Lys Glu Thr

465 470 475 480

Asn Arg Asp Glu Ser Phe Tyr Gly Asp Phe Val Leu Ala Tyr Asp Ile

485 490 495

Leu Leu Lys Val Asp His Ile Tyr Asp Ala Ile Arg Asn Tyr Val Thr

500 505 510

Gln Lys Pro Tyr Ser Lys Asp Lys Phe Lys Leu Tyr Phe Gln Asn Pro

515 520 525

Gln Phe Met Gly Gly Trp Asp Lys Asp Lys Glu Thr Asp Tyr Arg Ala

530 535 540

Thr Ile Leu Arg Tyr Gly Ser Lys Tyr Tyr Leu Ala Ile Met Asp Lys

545 550 555 560

Lys Tyr Ala Lys Cys Leu Gln Lys Ile Asp Lys Asp Asp Val Asn Gly

565 570 575

Asn Tyr Glu Lys Ile Asn Tyr Lys Leu Leu Pro Gly Pro Asn Lys Met

580 585 590

Leu Pro Lys Val Phe Phe Ser Lys Lys Trp Met Ala Tyr Tyr Asn Pro

595 600 605

Ser Glu Asp Ile Gln Lys Ile Tyr Lys Asn Gly Thr Phe Lys Lys Gly

610 615 620

Asp Met Phe Asn Leu Asn Asp Cys His Lys Leu Ile Asp Phe Phe Lys

625 630 635 640

Asp Ser Ile Ser Arg Tyr Pro Lys Trp Ser Asn Ala Tyr Asp Phe Asn

645 650 655

Phe Ser Glu Thr Glu Lys Tyr Lys Asp Ile Ala Gly Phe Tyr Arg Glu

660 665 670

Val Glu Glu Gln Gly Tyr Lys Val Ser Phe Glu Ser Ala Ser Lys Lys

675 680 685

Glu Val Asp Lys Leu Val Glu Glu Gly Lys Leu Tyr Met Phe Gln Ile

690 695 700

Tyr Asn Lys Asp Phe Ser Asp Lys Ser His Gly Thr Pro Asn Leu His

705 710 715 720

Thr Met Tyr Phe Lys Leu Leu Phe Asp Glu Asn Asn His Gly Gln Ile

725 730 735

Arg Leu Ser Gly Gly Ala Glu Leu Phe Met Arg Arg Ala Ser Leu Lys

740 745 750

Lys Glu Glu Leu Val Val His Pro Ala Asn Ser Pro Ile Ala Asn Lys

755 760 765

Asn Pro Asp Asn Pro Lys Lys Thr Thr Thr Leu Ser Tyr Asp Val Tyr

770 775 780

Lys Asp Lys Arg Phe Ser Glu Asp Gln Tyr Glu Leu His Ile Pro Ile

785 790 795 800

Ala Ile Asn Lys Cys Pro Lys Asn Ile Phe Lys Ile Asn Thr Glu Val

805 810 815

Arg Val Leu Leu Lys His Asp Asp Asn Pro Tyr Val Ile Gly Ile Ala

820 825 830

Arg Gly Glu Arg Asn Leu Leu Tyr Ile Val Val Val Asp Gly Lys Gly

835 840 845

Asn Ile Val Glu Gln Tyr Ser Leu Asn Glu Ile Ile Asn Asn Phe Asn

850 855 860

Gly Ile Arg Ile Lys Thr Asp Tyr His Ser Leu Leu Asp Lys Lys Glu

865 870 875 880

Lys Glu Arg Phe Glu Ala Arg Gln Asn Trp Thr Ser Ile Glu Asn Ile

885 890 895

Lys Glu Leu Lys Ala Gly Tyr Ile Ser Gln Val Val His Lys Ile Cys

900 905 910

Glu Leu Val Glu Lys Tyr Asp Ala Val Ile Ala Leu Ala Asp Leu Asn

915 920 925

Ser Gly Phe Lys Asn Ser Arg Val Lys Val Glu Lys Gln Val Tyr Gln

930 935 940

Lys Phe Glu Lys Met Leu Ile Asp Lys Leu Asn Tyr Met Val Asp Lys

945 950 955 960

Lys Ser Asn Pro Cys Ala Thr Gly Gly Ala Leu Lys Gly Tyr Gln Ile

965 970 975

Thr Asn Lys Phe Glu Ser Phe Lys Ser Met Ser Thr Gln Asn Gly Phe

980 985 990

Ile Phe Tyr Ile Pro Ala Trp Leu Thr Ser Lys Ile Asp Pro Ser Thr

995 1000 1005

Gly Phe Val Asn Leu Leu Lys Thr Lys Tyr Thr Ser Ile Ala Asp

1010 1015 1020

Ser Lys Lys Phe Ile Ser Ser Phe Asp Arg Ile Met Tyr Val Pro

1025 1030 1035

Glu Glu Asp Leu Phe Glu Phe Ala Leu Asp Tyr Lys Asn Phe Ser

1040 1045 1050

Arg Thr Asp Ala Asp Tyr Ile Lys Lys Trp Lys Leu Tyr Ser Tyr

1055 1060 1065

Gly Asn Arg Ile Arg Ile Phe Arg Asn Pro Lys Lys Asn Asn Val

1070 1075 1080

Phe Asp Trp Glu Glu Val Cys Leu Thr Ser Ala Tyr Lys Glu Leu

1085 1090 1095

Phe Asn Lys Tyr Gly Ile Asn Tyr Gln Gln Gly Asp Ile Arg Ala

1100 1105 1110

Leu Leu Cys Glu Gln Ser Asp Lys Ala Phe Tyr Ser Ser Phe Met

1115 1120 1125

Ala Leu Met Ser Leu Met Leu Gln Met Arg Asn Ser Ile Thr Gly

1130 1135 1140

Arg Thr Asp Val Asp Phe Leu Ile Ser Pro Val Lys Asn Ser Asp

1145 1150 1155

Gly Ile Phe Tyr Asp Ser Arg Asn Tyr Glu Ala Gln Glu Asn Ala

1160 1165 1170

Ile Leu Pro Lys Asn Ala Asp Ala Asn Gly Ala Tyr Asn Ile Ala

1175 1180 1185

Arg Lys Val Leu Trp Ala Ile Gly Gln Phe Lys Lys Ala Glu Asp

1190 1195 1200

Glu Lys Leu Asp Lys Val Lys Ile Ala Ile Ser Asn Lys Glu Trp

1205 1210 1215

Leu Glu Tyr Ala Gln Thr Ser Val Lys His

1220 1225

<210> 36

<211> 1314

<212> Protein

<213> Artificial sequence

<220>

<223> Synthetic polypeptide

<400> 36

Met Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Gly Ser Gly Met Ser

1. 5 10 15

Ile Tyr Gln Glu Phe Val Asn Lys Tyr Ser Leu Ser Lys Thr Leu Arg

20 25 30

Phe Glu Leu Ile Pro Gln Gly Lys Thr Leu Glu Asn Ile Lys Ala Arg

35 40 45

Gly Leu Ile Leu Asp Asp Glu Lys Arg Ala Lys Asp Tyr Lys Lys Ala

50 55 60

Lys Gln Ile Ile Asp Lys Tyr His Gln Phe Phe Ile Glu Glu Ile Leu

65 70 75 80

Ser Ser Val Cys Ile Ser Glu Asp Leu Leu Gln Asn Tyr Ser Asp Val

85 90 95

Tyr Phe Lys Leu Lys Lys Ser Asp Asp Asp Asn Leu Gln Lys Asp Phe

100 105 110

Lys Ser Ala Lys Asp Thr Ile Lys Lys Gln Ile Ser Glu Tyr Ile Lys

115 120 125

Asp Ser Glu Lys Phe Lys Asn Leu Phe Asn Gln Asn Leu Ile Asp Ala

130 135 140

Lys Lys Gly Gln Glu Ser Asp Leu Ile Leu Trp Leu Lys Gln Ser Lys

145 150 155 160

Asp Asn Gly Ile Glu Leu Phe Lys Ala Asn Ser Asp Ile Thr Asp Ile

165 170 175

Asp Glu Ala Leu Glu Ile Ile Lys Ser Phe Lys Gly Trp Thr Thr Tyr

180 185 190

Phe Lys Gly Phe His Glu Asn Arg Lys Asn Val Tyr Ser Ser Asn Asp

195 200 205

Ile Pro Thr Ser Ile Ile Tyr Arg Ile Val Asp Asp Asn Leu Pro Lys

210 215 220

Phe Leu Glu Asn Lys Ala Lys Tyr Glu Ser Leu Lys Asp Lys Ala Pro

225 230 235 240

Glu Ala Ile Asn Tyr Glu Gln Ile Lys Lys Asp Leu Ala Glu Glu Leu

245 250 255

Thr Phe Asp Ile Asp Tyr Lys Thr Ser Glu Val Asn Gln Arg Val Phe

260 265 270

Ser Leu Asp Glu Val Phe Glu Ile Ala Asn Phe Asn Asn Tyr Leu Asn

275 280 285

Gln Ser Gly Ile Thr Lys Phe Asn Thr Ile Ile Gly Gly Lys Phe Val

290 295 300

Asn Gly Glu Asn Thr Lys Arg Lys Gly Ile Asn Glu Tyr Ile Asn Leu

305 310 315 320

Tyr Ser Gln Gln Ile Asn Asp Lys Thr Leu Lys Lys Tyr Lys Met Ser

325 330 335

Val Leu Phe Lys Gln Ile Leu Ser Asp Thr Glu Ser Lys Ser Phe Val

340 345 350

Ile Asp Lys Leu Glu Asp Asp Ser Asp Val Val Thr Thr Met Gln Ser

355 360 365

Phe Tyr Glu Gln Ile Ala Ala Phe Lys Thr Val Glu Glu Lys Ser Ile

370 375 380

Lys Glu Thr Leu Ser Leu Leu Phe Asp Asp Leu Lys Ala Gln Lys Leu

385 390 395 400

Asp Leu Ser Lys Ile Tyr Phe Lys Asn Asp Lys Ser Leu Thr Asp Leu

405 410 415

Ser Gln Gln Val Phe Asp Asp Tyr Ser Val Ile Gly Thr Ala Val Leu

420 425 430

Glu Tyr Ile Thr Gln Gln Ile Ala Pro Lys Asn Leu Asp Asn Pro Ser

435 440 445

Lys Lys Glu Gln Glu Leu Ile Ala Lys Lys Thr Glu Lys Ala Lys Tyr

450 455 460

Leu Ser Leu Glu Thr Ile Lys Leu Ala Leu Glu Glu Phe Asn Lys His

465 470 475 480

Arg Asp Ile Asp Lys Gln Cys Arg Phe Glu Glu Ile Leu Ala Asn Phe

485 490 495

Ala Ala Ile Pro Met Ile Phe Asp Glu Ile Ala Gln Asn Lys Asp Asn

500 505 510

Leu Ala Gln Ile Ser Ile Lys Tyr Gln Asn Gln Gly Lys Lys Asp Leu

515 520 525

Leu Gln Ala Ser Ala Glu Asp Asp Val Lys Ala Ile Lys Asp Leu Leu

530 535 540

Asp Gln Thr Asn Asn Leu Leu His Lys Leu Lys Ile Phe His Ile Ser

545 550 555 560

Gln Ser Glu Asp Lys Ala Asn Ile Leu Asp Lys Asp Glu His Phe Tyr

565 570 575

Leu Val Phe Glu Glu Cys Tyr Phe Glu Leu Ala Asn Ile Val Pro Leu

580 585 590

Tyr Asn Lys Ile Arg Asn Tyr Ile Thr Gln Lys Pro Tyr Ser Asp Glu

595 600 605

Lys Phe Lys Leu Asn Phe Glu Asn Ser Thr Leu Ala Asn Gly Trp Asp

610 615 620

Lys Asn Lys Glu Pro Asp Asn Thr Ala Ile Leu Phe Ile Lys Asp Asp

625 630 635 640

Lys Tyr Tyr Leu Gly Val Met Asn Lys Lys Asn Asn Lys Ile Phe Asp

645 650 655

Asp Lys Ala Ile Lys Glu Asn Lys Gly Glu Gly Tyr Lys Lys Ile Val

660 665 670

Tyr Lys Leu Leu Pro Gly Ala Asn Lys Met Leu Pro Lys Val Phe Phe

675 680 685

Ser Ala Lys Ser Ile Lys Phe Tyr Asn Pro Ser Glu Asp Ile Leu Arg

690 695 700

Ile Arg Asn His Ser Thr His Thr Lys Asn Gly Ser Pro Gln Lys Gly

705 710 715 720

Tyr Glu Lys Phe Glu Phe Asn Ile Glu Asp Cys Arg Lys Phe Ile Asp

725 730 735

Phe Tyr Lys Gln Ser Ile Ser Lys His Pro Glu Trp Lys Asp Phe Gly

740 745 750

Phe Arg Phe Ser Asp Thr Gln Arg Tyr Asn Ser Ile Asp Glu Phe Tyr

755 760 765

Arg Glu Val Glu Asn Gln Gly Tyr Lys Leu Thr Phe Glu Asn Ile Ser

770 775 780

Glu Ser Tyr Ile Asp Ser Val Val Asn Gln Gly Lys Leu Tyr Leu Phe

785 790 795 800

Gln Ile Tyr Asn Lys Asp Phe Ser Ala Tyr Ser Lys Gly Arg Pro Asn

805 810 815

Leu His Thr Leu Tyr Trp Lys Ala Leu Phe Asp Glu Arg Asn Leu Gln

820 825 830

Asp Val Val Tyr Lys Leu Asn Gly Glu Ala Glu Leu Phe Tyr Arg Lys

835 840 845

Gln Ser Ile Pro Lys Lys Ile Thr His Pro Ala Lys Glu Ala Ile Ala

850 855 860

Asn Lys Asn Lys Asp Asn Pro Lys Lys Glu Ser Val Phe Glu Tyr Asp

865 870 875 880

Leu Ile Lys Asp Lys Arg Phe Thr Glu Asp Lys Phe Phe Phe His Cys

885 890 895

Pro Ile Thr Ile Asn Phe Lys Ser Ser Gly Ala Asn Lys Phe Asn Asp

900 905 910

Glu Ile Asn Leu Leu Leu Lys Glu Lys Ala Asn Asp Val His Ile Leu

915 920 925

Ser Ile Ala Arg Gly Glu Arg His Leu Ala Tyr Tyr Thr Leu Val Asp

930 935 940

Gly Lys Gly Asn Ile Ile Lys Gln Asp Thr Phe Asn Ile Ile Gly Asn

945 950 955 960

Asp Arg Met Lys Thr Asn Tyr His Asp Lys Leu Ala Ala Ile Glu Lys

965 970 975

Asp Arg Asp Ser Ala Arg Lys Asp Trp Lys Lys Ile Asn Asn Ile Lys

980 985 990

Glu Met Lys Glu Gly Tyr Leu Ser Gln Val Val His Glu Ile Ala Lys

995 1000 1005

Leu Val Ile Glu Tyr Asn Ala Ile Val Val Phe Glu Asp Leu Asn

1010 1015 1020

Phe Gly Phe Lys Arg Gly Arg Phe Lys Val Glu Lys Gln Val Tyr

1025 1030 1035

Gln Lys Leu Glu Lys Met Leu Ile Glu Lys Leu Asn Tyr Leu Val

1040 1045 1050

Phe Lys Asp Asn Glu Phe Asp Lys Thr Gly Gly Val Leu Arg Ala

1055 1060 1065

Tyr Gln Leu Thr Ala Pro Phe Glu Thr Phe Lys Lys Met Gly Lys

1070 1075 1080

Gln Thr Gly Ile Ile Tyr Tyr Val Pro Ala Gly Phe Thr Ser Lys

1085 1090 1095

Ile Cys Pro Val Thr Gly Phe Val Asn Gln Leu Tyr Pro Lys Tyr

1100 1105 1110

Glu Ser Val Ser Lys Ser Gln Glu Phe Phe Ser Lys Phe Asp Lys

1115 1120 1125

Ile Cys Tyr Asn Leu Asp Lys Gly Tyr Phe Glu Phe Ser Phe Asp

1130 1135 1140

Tyr Lys Asn Phe Gly Asp Lys Ala Ala Lys Gly Lys Trp Thr Ile

1145 1150 1155

Ala Ser Phe Gly Ser Arg Leu Ile Asn Phe Arg Asn Ser Asp Lys

1160 1165 1170

Asn His Asn Trp Asp Thr Arg Glu Val Tyr Pro Thr Lys Glu Leu

1175 1180 1185

Glu Lys Leu Leu Lys Asp Tyr Ser Ile Glu Tyr Gly His Gly Glu

1190 1195 1200

Cys Ile Lys Ala Ala Ile Cys Gly Glu Ser Asp Lys Lys Phe Phe

1205 1210 1215

Ala Lys Leu Thr Ser Val Leu Asn Thr Ile Leu Gln Met Arg Asn

1220 1225 1230

Ser Lys Thr Gly Thr Glu Leu Asp Tyr Leu Ile Ser Pro Val Ala

1235 1240 1245

Asp Val Asn Gly Asn Phe Phe Asp Ser Arg Gln Ala Pro Lys Asn

1250 1255 1260

Met Pro Gln Asp Ala Asp Ala Asn Gly Ala Tyr His Ile Gly Leu

1265 1270 1275

Lys Gly Leu Met Leu Leu Gly Arg Ile Lys Asn Asn Gln Glu Gly

1280 1285 1290

Lys Lys Leu Asn Leu Val Ile Lys Asn Glu Glu Tyr Phe Glu Phe

1295 1300 1305

Val Gln Asn Arg Asn Asn

1310

<210> 37

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 37

actgcggaaa tttgagcgt 19

<210> 38

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 38

acgctcaaat ttccgcagt 19

<210> 39

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 39

aggcaatggc tgcacatgc 19

<210> 40

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 40

gcatgtgcag ccattgcct 19

<210> 41

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 41

gacgcttggt tctgaggag 19

<210> 42

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 42

ctcctcagaa ccaagcgtc 19

<210> 43

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 43

tccggaaacg cattcctct 19

<210> 44

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 44

agaggaatgc gtttccgga 19

<210> 45

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 45

ccgcgtcagc ccggcccgg 19

<210> 46

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 46

ccgggccggg ctgacgcgg 19

<210> 47

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 47

cgactcccgc tgggcctct 19

<210> 48

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 48

agaggcccag cgggagtcg 19

<210> 49

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 49

ccgttgcgcg ctcgctctc 19

<210> 50

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 50

gagagcgagc gcgcaacgg 19

<210> 51

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 51

ccgcgcatcc tgccaggcc 19

<210> 52

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 52

ggcctggcag gatgcgcgg 19

<210> 53

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 53

ccaacttggc gcgtttcgg 19

<210> 54

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 54

ccgaaacgcg ccaagttgg 19

<210> 55

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 55

accacgcgtc cgagtccgg 19

<210> 56

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 56

ccggactcgg acgcgtggt 19

<210> 57

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 57

tgctcattgt ccctggaca 19

<210> 58

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 58

tgtccaggga caatgagca 19

<210> 59

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 59

ggacaccctg ctcattgtc 19

<210> 60

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 60

gacaatgagc agggtgtcc 19

<210> 61

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 61

accggcagcc tgcgcgtcc 19

<210> 62

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 62

ggacgcgcag gctgccggt 19

<210> 63

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 63

cgatgggcac ccactgctc 19

<210> 64

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 64

gagcagtggg tgcccatcg 19

<210> 65

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 65

ccttcacgtg gacgcgcag 19

<210> 66

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 66

ctgcgcgtcc acgtgaagg 19

<210> 67

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 67

cgtgaaggtg gaagccttc 19

<210> 68

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 68

gaaggcttcc accttcacg 19

<210> 69

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 69

ctccttggtc aggcgccgg 19

<210> 70

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 70

ccggcgcctg accaaggag 19

<210> 71

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 71

tggtcaggcg ccggttccg 19

<210> 72

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 72

cggaaccggc gcctgacca 19

<210> 73

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 73

tagaggtcgc cttctcctc 19

<210> 74

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 74

gaggagaagg cgacctcta 19

<210> 75

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 75

cgacgctcgg gtcgcggtg 19

<210> 76

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 76

caccgcgacc cgagcgtcg 19

<210> 77

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 77

atgctgtcgc cgcgcgggg 19

<210> 78

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 78

ccccgcgcgg cgacagcat 19

<210> 79

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 79

ctcaccctca ccggagcca 19

<210> 80

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 80

tggctccggt gagggtgag 19

<210> 81

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 81

ccgcaaactt tactcctta 19

<210> 82

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 82

taaggagtaaagtttgcgg 19

<210> 83

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 83

ctcctaagat tggcttcac 19

<210> 84

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 84

gtgaagccaa tcttaggag 19

<210> 85

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 85

ccggagccac tcctaagat 19

<210> 86

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 86

atcttaggag tggctccgg 19

<210> 87

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 87

ttctctacct tacgtctca 19

<210> 88

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 88

tgagacgtag ggtagagaa 19

<210> 89

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 89

tacgtctcat tctccgcaa 19

<210> 90

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 90

ttgcggagaa tgagacgta 19

<210> 91

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 91

gctaggcctc cagcccttc 19

<210> 92

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 92

gaagggctgg aggcctagc 19

<210> 93

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 93

acaggtggcg ccgcaactt 19

<210> 94

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 94

aagttgcggc gccacctgt 19

<210> 95

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 95

agccggaggc gcgagagtc 19

<210> 96

<211> 19

<212> DNA

<213> Artificial sequence

<220>

<223> Synthetic polynucleotide

<400> 96

gactctcgcg cctccggct 19

<---

Claims (65)

1. A fusion protein for silencing a target nucleic acid sequence, containing, in the direction from the N-terminus to the C-terminus: (i) DNA methyltransferase domain, nuclease-deficient RNA-guided DNA endonuclease enzyme, and box-associated domain ; or (ii) the domain associated with the box , a nuclease-deficient RNA-guided DNA endonuclease enzyme and a DNA methyltransferase domain. 2. The fusion protein of claim 1, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is a class II nuclease-deficient CRISPR endonuclease. 3. The fusion protein according to claim 1, characterized in that said DNA methyltransferase domain contains a Dnmt3A protein and a Dnmt3L protein (Dnmt3A-3L domain). 4. The fusion protein of claim 1, wherein said fusion protein includes, from N-terminus to C-terminus, a DNA methyltransferase domain, an enzyme-deficient RNA-guided DNA endonuclease, and a box-linked domain . 5. The fusion protein according to claim 4, characterized in that said nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9, and said DNA methyltransferase domain contains a Dnmt3A protein and a Dnmt3L protein (Dnmt3A-3L domain); wherein said dCas9 is covalently linked to the Dnmt3A-3L domain via an XTEN linker. 6. The fusion protein of claim 5, wherein said dCas9 is covalently linked to said Dnmt3A-3L domain via an XTEN linker, and wherein said Dnmt3A-3L domain is covalently linked to said box-linked domain , via the XTEN linker. 7. The fusion protein according to claim 1, characterized in that said fusion protein includes, in the direction from the N-terminus to the C-terminus, a box-linked domain , a nuclease-deficient RNA-guided DNA endonuclease enzyme, an XTEN linker, and a DNA methyltransferase domain. 8. The fusion protein according to claim 7, characterized in that said nuclease-deficient RNA-guided DNA endonuclease enzyme is dCas9, and said DNA methyltransferase domain contains a Dnmt3A protein and a Dnmt3L protein (Dnmt3A-3L domain). 9. The fusion protein according to claim 8, characterized in that said dCas9 is covalently linked to said Dnmt3A-3L domain via an XTEN linker, and wherein said domain is associated with a box , is covalently linked to the indicated dCas9 via the XTEN linker. 10. The fusion protein of claim 1, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is covalently linked to said box-linked domain , via a peptide linker. 11. The fusion protein of claim 1, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is covalently linked to said DNA methyltransferase domain via a peptide linker. 12. The fusion protein according to claim 1, characterized in that said domain associated with the box , is covalently linked to the indicated DNA methyltransferase domain via the XTEN linker. 13. The fusion protein of claim 10, wherein said peptide linker is an XTEN linker. 14. The fusion protein of claim 13, wherein said XTEN linker contains from about 16 to about 80 amino acid residues. 15. The fusion protein according to claim 1, additionally containing a nuclear localization signal peptide. 16. The fusion protein according to claim 1, characterized in that said fusion protein contains an amino acid sequence with SEQ ID NO: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15. 17. Nucleic acid encoding the fusion protein according to claim 1. 18. Nucleic acid according to claim 17, characterized in that said nucleic acid sequence is messenger RNA. 19. A complex for silencing a target nucleic acid sequence, containing: (i) a fusion protein according to claim 1; and (ii) a polynucleotide comprising: (a) a DNA targeting sequence complementary to the target polynucleotide sequence; and (b) a binding sequence for said nuclease-deficient RNA-directed DNA endonuclease enzyme, wherein said nuclease-deficient RNA-directed DNA endonuclease enzyme is linked to said polynucleotide through a binding sequence. 20. The complex according to claim 19, characterized in that said target polynucleotide sequence is part of a gene. 21. The complex according to claim 19, characterized in that said target polynucleotide sequence is part of a transcription regulatory sequence. 22. The complex according to claim 19, characterized in that said target polynucleotide sequence is part of a promoter, enhancer or silencer. 23. The complex according to claim 19, characterized in that said target polynucleotide sequence is within about 3000 bp. and flanks the transcription start site. 24. An expression vector for silencing the target gene, containing a nucleic acid sequence encoding the fusion protein according to claim 1. 25. The expression vector of claim 24, further comprising a polynucleotide, characterized in that said polynucleotide contains: (a) a DNA targeting sequence that is complementary to the target polynucleotide sequence; and (b) a binding sequence for the enzyme nuclease-deficient RNA-guided DNA endonuclease. 26. Selected cell for targeted gene silencing, containing the fusion protein according to claim 1; nucleic acid according to claim 17; the complex according to claim 19 or the vector according to claim 24, where the specified cell contains the target gene that has undergone silencing. 27. The isolated cell according to claim 26, characterized in that said cell is a eukaryotic cell. 28. The isolated cell of claim 26, wherein said cell is a mammalian cell. 29. A method for silencing a target nucleic acid sequence in a cell, comprising: (i) delivering a first polynucleotide encoding the fusion protein of claim 1 into a cell containing said target nucleic acid; and (ii) delivering to said cell a second polynucleotide that includes (a) a DNA targeting sequence that is complementary to said target nucleic acid sequence, and (b) a binding sequence for said nuclease-deficient RNA-guided DNA endonuclease enzyme. 30. The method according to claim 29, characterized in that said fusion protein causes silencing of said target nucleic acid sequence in said cell by methylating chromatin containing said target nucleic acid sequence and/or by introducing repressive chromatin markers into chromatin containing said target nucleic acid sequence. 31. The method according to claim 29, characterized in that said first polynucleotide is contained in a first vector. 32. The method according to claim 29, characterized in that said first polynucleotide is contained in a second vector. 33. The method according to claim 32, characterized in that said first vector and second vector are the same. 34. The method according to claim 32, characterized in that said first vector is different from said second vector. 35. The method according to claim 29, characterized in that said cell is a mammalian cell. 36. The method according to claim 29, characterized in that the specificity of this method is 2 times higher than the specificity with respect to the sequence of the non-target nucleic acid. 37. A method for silencing a target nucleic acid sequence in a cell, comprising delivering the complex according to claim 19 into a cell containing said target nucleic acid. 38. The method according to claim 37, characterized in that the specified complex causes the silencing of the specified sequence of the target nucleic acid in the specified cell by methylation of the chromatin containing the specified sequence of the target nucleic acid, and (or) by introducing repressive chromatin markers into the chromatin containing the specified sequence target nucleic acid. 39. The method according to claim 37, characterized in that said cell is a mammalian cell. 40. The method according to claim 37, characterized in that the specificity of this method is 2 times higher than the specificity with respect to the sequence of the non-target nucleic acid. 41. The fusion protein according to claim 1, characterized in that said nuclease-deficient RNA-guided DNA endonuclease enzyme is a dCas9 enzyme that contains an amino acid sequence having at least 85% sequence identity with SEQ ID NO:23; ddCas12a, which contains an amino acid sequence at least 85% identical to SEQ ID NO:34; dLbCfp1, which contains an amino acid sequence at least 85% identical to SEQ ID NO:35; or dFnCfp1, which contains an amino acid sequence that is at least 85% identical to SEQ ID NO: 36. 42. The fusion protein according to claim 1, characterized in that said domain associated with the box , contains an amino acid sequence having at least 85% sequence identity with SEQ ID NO: 16. 43. The fusion protein of claim 3, wherein said Dnmt3A protein contains an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 26, and said Dnmt3L protein contains an amino acid sequence having at least 85% sequence identity sequences with SEQ ID NO: 28. 44. The fusion protein of claim 1, wherein said fusion protein contains an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 15. 45. The fusion protein according to claim 1, wherein said nuclease-deficient RNA-guided DNA endonuclease enzyme is selected from the group consisting of dCas9, ddCpf1, ddCas12a, ddLbCfp1 and ddFnCfp1. 46. A fusion protein for silencing a target nucleic acid sequence containing a DNA methyltransferase domain, a nuclease-deficient RNA-guided DNA endonuclease enzyme, and a box-linked domain ; Where (1) said nuclease-deficient RNA-guided DNA endonuclease enzyme is a dCas9 enzyme that contains an amino acid sequence having at least 85% sequence identity with SEQ ID NO:23; ddCas12a, which contains an amino acid sequence at least 85% identical to SEQ ID NO:34; dLbCfp1, which contains an amino acid sequence at least 85% identical to SEQ ID NO:35; or dFnCfp1, which contains an amino acid sequence at least 85% identical to SEQ ID NO: 36; (2) the specified domain associated with the box , contains an amino acid sequence having at least 85% sequence identity with SEQ ID NO: 16; And (3) the DNA methyltransferase domain contains a Dnmt3A protein, a Dnmt3L protein, or a Dnmt3A protein and a Dnmt3L protein; wherein said Dnmt3A protein contains an amino acid sequence having at least 85% sequence identity with SEQ ID NO: 26, and said Dnmt3L protein contains an amino acid sequence having at least 85% sequence identity with SEQ ID NO: 28. 47. The fusion protein according to claim 46, characterized in that said box-linked domain , contains an amino acid sequence having at least 90% sequence identity with SEQ ID NO: 16. 48. The fusion protein according to claim 47, characterized in that said box-linked domain , contains an amino acid sequence having at least 95% sequence identity with SEQ ID NO: 16. 49. The fusion protein according to claim 48, characterized in that said box-linked domain , contains an amino acid sequence having at least 99% sequence identity with SEQ ID NO: 16. 50. The fusion protein of claim 46, wherein said Dnmt3A protein contains an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 26. 51. The fusion protein of claim 50, wherein said Dnmt3A protein contains an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 26. 52. The fusion protein of claim 51, wherein said Dnmt3A protein contains an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 26. 53. The fusion protein of claim 46, wherein said Dnmt3L protein contains an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 28. 54. The fusion protein of claim 53, wherein said Dnmt3L protein contains an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 28. 55. The fusion protein of claim 54, wherein said Dnmt3L protein contains an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 28. 56. The fusion protein of claim 46, wherein said dCas9 contains an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 23. 57. The fusion protein of claim 56, wherein said dCas9 contains an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 23. 58. The fusion protein of claim 57, wherein said dCas9 contains an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 23. 59. The fusion protein according to claim 46, characterized in that said box-linked domain , contains the amino acid sequence of SEQ ID NO: 16, the specified Dnmt3A protein contains the amino acid sequence of SEQ ID NO: 26, the specified Dnmt3L protein contains the amino acid sequence of SEQ ID NO: 28, and the specified dCas9 contains the amino acid sequence of SEQ ID NO: 23. 60. The fusion protein of claim 59, wherein said fusion protein comprises, from N-terminus to C-terminus, a DNA methyltransferase domain, a nuclease-deficient RNA-guided DNA endonuclease enzyme, and a box-linked domain .