阅读说明：本技术 Cas蛋白抑制剂的组合物及应用 (Cas protein inhibitor composition and application ) 是由 刘佳 姜标 马培翔 杨光 于 2019-08-29 设计创作，主要内容包括：本发明提供了能够结合并抑制Cas蛋白与核酸分子结合的能力的氨基酸序列,从而抑制所述Cas蛋白在基因组编辑中的功能。此类Cas蛋白抑制剂可由主要外壳蛋白(G8P)、所述G8P的胞外区(G8P-(EX))或生物等效物组成,可用于改善基于Cas蛋白的基因组编辑过程的特异性。(The present invention provides amino acid sequences capable of binding to and inhibiting the ability of a Cas protein to bind to a nucleic acid molecule, thereby inhibiting the function of the Cas protein in genome editing. Such Cas protein inhibitors may consist of a major coat protein (G8P), the extracellular region of said G8P (G8P) EX ) Or bioequivalent compositions, useful for improving the specificity of Cas protein-based genome editing processes.)

1. A method of improving the specificity of a Cas protein-based genome editing process, the method comprising contacting a sample subjected to the Cas protein-based genome editing process with a polypeptide or a polynucleotide encoding the polypeptide, wherein the polypeptide comprises a major coat protein (G8P) capable of binding to the Cas protein, an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

2. The method of claim 1, wherein the Cas protein is selected from the group consisting of SpCas9, SaCas9, NmeCas9, StCas9, CjCas9, AsCpf1, FnCpf1, SsCpf1, PcCpf1, BpCpf1, CmtCpf1, LiCpf1, PmCpf1, Pb3310Cpf1, Pb4417Cpf1, BsCpf1, EeCpf1, RfCas13d, lwascas 13 Cas13a, PspCas13b, PguCas13b, RanCas13b, variants thereof, and chemically modified forms thereof, said Cas protein being capable of interacting with said chemically modified polypeptide.

3. The method of claim 1 or 2, wherein the Cas protein and the polypeptide or polynucleotide are provided simultaneously to a sample.

4. The method of claim 3, wherein said polynucleotide further comprises an inducible promoter.

5. The method of claim 1 or 2, wherein the polypeptide or polynucleotide is provided after the sample has been contacted with the Cas protein.

6. The method of any one of claims 1-5, wherein the polypeptide is chemically modified.

7. The method of any one of claims 1-6, wherein the Cas protein-based genome editing process is performed in vitro.

8. The method of any one of claims 1-6, wherein the Cas protein-based genome editing process is performed in a living subject.

9. The method of claim 8, wherein the living subject is a human subject.

10. The method of any one of claims 1-9, wherein the polypeptide is delivered as part of a viral particle or nanoparticle.

11. The method of claim 10, wherein the viral particle comprises M13 bacteriophage.

12. The method of any one of claims 1-11, wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs 1-22 and 37-436, or a sequence having at least 70% sequence identity to any one of SEQ ID NOs 1-22 and 37-436, and is capable of inhibiting binding to a Cas protein.

13. The method of claim 12, wherein said polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs 1-10 and 21.

14. A method of genome editing in a subject, the method comprising:

administering a Cas protein-based genome editing system to a subject; and thereafter

Administering to a subject a polypeptide or a polynucleotide encoding the polypeptide, wherein the polypeptide comprises a major coat protein capable of binding the Cas protein (G8P), an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

15. The method of claim 14, wherein the polypeptide or polynucleotide is administered after initiation of genome editing of the Cas protein-based genome editing system.

16. The method of claim 14, wherein the polypeptide or polynucleotide is administered at least 12 hours after administration of the Cas protein-based genome editing system.

17. The method of any one of claims 14-16, wherein the administration is intravenous, intramuscular, nasal spray, or topical.

18. A method of genome editing in a subject, the method comprising administering to the subject a Cas protein-based genome editing system and a polynucleotide encoding a polypeptide operably linked to an inducible promoter, wherein the polypeptide comprises a major coat protein (G8P) capable of binding the Cas protein, an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

19. The method of claim 18, further comprising inducing expression of the polypeptide by activating the inducible promoter after initiation of genome editing of the Cas protein-based genome editing system.

20. The method of claim 18 or 19, wherein the Cas protein and the polypeptide are encoded by the same nucleic acid construct.

21. A recombinant expression vector comprising a first polynucleotide fragment and a second polynucleotide fragment, the first polynucleotide fragment encoding a Cas protein; the second polynucleotide fragment encodes a polypeptide comprising a major coat protein capable of binding the Cas protein (G8P), an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA polypeptide of the bioequivalence of (1).

22. The vector of claim 21, wherein said second polynucleotide segment is operably linked to an inducible promoter for expression of said polypeptide in a cell.

23. The vector of claim 21, wherein the Cas protein is selected from the group consisting of SpCas9, SaCas9, NmeCas9, StCas9, CjCas9, AsCpf1, FnCpf1, SsCpf1, PcCpf1, BpCpf1, CmtCpf1, LiCpf1, PmCpf1, Pb3310Cpf1, Pb4417Cpf1, BsCpf1, EeCpf1, RfCas13d, lwascas 13 Cas13a, PspCas13b, PguCas13b, RanCas13b, and variants thereof.

24. The vector of any one of claims 21-23, further comprising a coding sequence for one or more proteins selected from the group consisting of cytidine deaminase and Uracil Glycosylase Inhibitor (UGI).

25. A recombinant expression vector comprising a nucleotide fragment encoding a polypeptide and an operably linked promoter for expression of an amino acid sequence in a eukaryotic cell, wherein the polypeptide comprises a major coat protein capable of binding to the Cas protein (G8P), an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

26. The vector of claim 25, wherein said polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NOs 1-22 and 37-436.

27. The vector of claim 25 or 26, wherein said promoter promotes transcription of said nucleotide fragment in a mammalian cell.

28. The vector of any one of claims 25 to 27, wherein the promoter is inducible.

29. A composition, combination or kit comprising a Cas protein and a polypeptide comprising a major coat protein (G8P) capable of binding to the Cas protein, an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

30. A molecule comprising a Cas protein, a polypeptide comprising a cleavable linker capable of linking the Cas protein and the polypeptide, and a cleavable linker connecting the Cas protein and the polypeptideA major coat protein (G8P) that binds the Cas protein, an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

31. The molecule of claim 30, wherein said cleavable linker is a peptide comprising a proteolytic cleavage site.

32. The molecule of claim 31, wherein said proteolytic cleavage site is a self-cleavage site.

33. The molecule of claim 30, wherein said cleavable linker is light-activated or drug-activated.

34. The molecule of any one of claims 30-33, wherein said Cas protein is fused to a cytosine deaminase or an adenosine deaminase.

35. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a polypeptide comprising a major coat protein (G8P) capable of binding to the Cas protein, the extracellular region of G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

36. The pharmaceutical composition of claim 35, wherein the polypeptide is delivered as part of a viral particle or nanoparticle.

37. The pharmaceutical composition of claim 36, wherein the viral particle comprises M13 bacteriophage.

38. The pharmaceutical composition of any one of claims 35-37, wherein the composition is provided in the form of an injection, tablet, capsule, gel, cream, or spray.

39. The method of any one of claims 1 to 20, the vector of any one of claims 21 to 28, the composition of claim 29, the molecule of any one of claims 30 to 34, the composition of any one of claims 35 to 37, wherein G8P is selected from the group consisting of SEQ ID NOs 11 to 20.

40. The method of any one of claims 1 to 20, the vector of any one of claims 21 to 28, the composition of claim 29, the molecule of any one of claims 30 to 34, the composition of any one of claims 35 to 37, wherein G8P is present_EXSelected from the group consisting of SEQ ID NO 1-10.

41. The method of any one of claims 1 to 20, the vector of any one of claims 21 to 28, the composition of claim 29, the molecule of any one of claims 30 to 34, the composition of any one of claims 35 to 37, wherein said bioequivalent is said G8P or said G8P_EXHas at least 70% sequence identity.

42. A polypeptide comprising an amino acid sequence comprising an addition, deletion, substitution, or combination thereof of one, two, three, four, or five amino acids in a sequence selected from the group consisting of SEQ ID NOs 1-22 and 37-436, wherein the polypeptide is capable of binding to a Cas protein.

43. A method of genome editing in a subject, the method comprising:

administering a Cas protein-based genome editing system to a subject; and

administering to the subject the polypeptide of claim 42 or a polynucleotide encoding the polypeptide.

Background

Genome editing can be used to correct driver mutations that are hidden behind genetic diseases, thereby enabling a complete cure for these diseases in living organisms; genome editing can also be used to engineer the genome of a crop, thereby increasing the yield of the crop and conferring resistance to environmental contamination or pathogen infection to the crop; also, microbial genome transformation by precise genome editing has important implications for the development of renewable biological energy sources.

Due to its unrivaled editing efficiency, convenience and potential application in living organisms, the CRISPR/Cas (regularly interspaced clustered short palindromic repeats/CRISPR-associated proteins) system has become the most powerful genome editing tool. Under the guidance of guide RNA (gridea RNA, gRNA), Cas nucleases can generate DNA Double Strand Breaks (DSB) at target genomic sites of various cells (cell lines and cells of living organisms). These DSBs are then repaired by endogenous DNA repair systems and can be used to perform the desired genome editing.

Recently, a Base Editor (BE) integrating a CRISPR/Cas system with an APOBEC (apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like) cytosine deaminase family was developed, which greatly improves the efficiency of CRISPR/Cas 9-mediated gene modification. Cytosine (C) deamination activity of rat APOBEC1(rA1) can be directed to target bases in the genome by fusion with Cas9 nickase (nCas9) or catalytically inactive Cas9(dCas9) and catalyze C replacement to thymine (T) at these bases.

During the use of genome editing methods, non-specific and unintended ("off-target") genetic modifications can occur. For example, in a CRISPR/Cas system, if the complexes do not bind to the target sequence (usually as a result of homologous sequences and/or mismatch tolerance), they will cleave off-target DSBs and cause non-specific genetic modifications. Off-target effects include unwanted point mutations, deletions, insertions, inversions, and translocations. There is a need to develop methods to reduce such off-target gene modifications.

Disclosure of Invention

The present disclosure provides compositions of peptide inhibitors for Cas proteins and their use in genome editing. One embodiment of the present disclosure provides a method of improving the specificity of a Cas protein-based genome editing process, the method comprising contacting a sample that has undergone a Cas protein-based genome editing process with a polypeptide or a polynucleotide encoding the polypeptide, wherein the polypeptide comprises a major coat protein (G8P) capable of binding to the Cas protein, an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

In some embodiments, the Cas protein is selected from the group consisting of SpCas9, SaCas9, NmeCas9, StCas9, CjCas9, AsCpf1, FnCpf1, SsCpf1, PcCpf1, BpCpf1, CmtCpf1, LiCpf1, PmCpf1, Pb3310Cpf1, Pb4417Cpf1, BsCpf1, EeCpf1, RfCas13d, LwaCas13a, PspCas13b, pgcas 13b, RanCas13b, variants thereof, and chemically modified forms thereof, said Cas protein being capable of interacting with said polypeptide inhibitor or chemically modified forms thereof. In some embodiments, the Cas protein and the polypeptide or polynucleotide are provided simultaneously to a sample.

In some embodiments, the polynucleotide further comprises an inducible promoter. In some embodiments, the polypeptide or polynucleotide is provided after the sample has been contacted with the Cas protein. In some embodiments, the polypeptide is chemically modified.

In some embodiments, the Cas protein-based genome editing process is performed in vitro. In some embodiments, the Cas protein-based genome editing process is performed in a living subject. In some embodiments, the living subject is, but is not limited to, a human subject, an animal subject, a plant subject, a yeast subject, a bacterial subject, or a viral subject.

In one embodiment, there is also provided a method of genome editing in a subject, the method comprising: administering a Cas protein-based genome editing system to a subject; and thereafter administering to the subject a polypeptide or a polynucleotide encoding the polypeptide, wherein the polypeptide comprises a major coat protein capable of binding the Cas protein (G8P), an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

In some embodiments, the polypeptide or polynucleotide is administered after initiation of genome editing of the Cas protein-based genome editing system. In some embodiments, the polypeptide or polynucleotide is administered at least 12 hours after administration of the Cas protein-based genome editing system. In some embodiments, the administration is intravenous, intramuscular, nasal spray, or topical administration.

Another embodiment provides a method of genome editing in a subject, the method comprising administering to the subject a Cas protein-based genome editing system and a polynucleotide encoding a polypeptide operably linked to an inducible promoter, wherein the polypeptide comprises a major coat protein (G8P) capable of binding the Cas protein, an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

In some embodiments, the method further comprises inducing expression of the polypeptide by activating the inducible promoter after initiation of genome editing of the Cas protein-based genome editing system. In some embodiments, the Cas protein and the polypeptide are encoded by the same nucleic acid construct.

In one embodiment, furtherA recombinant expression vector is provided comprising a first polynucleotide fragment encoding a Cas protein and a second polynucleotide fragment; the second polynucleotide fragment encodes a polypeptide comprising a major coat protein capable of binding the Cas protein (G8P), an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA polypeptide of the bioequivalence of (1).

In some embodiments, the second polynucleotide fragment is operably linked to an inducible promoter for expression of the polypeptide in a cell. In some embodiments, the Cas protein is selected from the group consisting of SpCas9, SaCas9, NmeCas9, StCas9, CjCas9, AsCpf1, FnCpf1, SsCpf1, PcCpf1, BpCpf1, CmtCpf1, LiCpf1, PmCpf1, Pb3310Cpf1, Pb4417Cpf1, BsCpf1, EeCpf1, RfCas13d, LwaCas13a, PspCas13b, pgcas 13b, RanCas13b, and variants thereof.

In some embodiments, the vector further comprises a coding sequence for one or more proteins selected from the group consisting of cytidine deaminase and Uracil Glycosylase Inhibitor (UGI).

Another embodiment provides a recombinant expression vector comprising a nucleotide fragment encoding a polypeptide and an operably linked promoter for expression of an amino acid sequence in a eukaryotic cell, wherein the polypeptide comprises a major coat protein capable of binding to the Cas protein (G8P), an extracellular region of the G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

In some embodiments, the promoter initiates transcription of the nucleotide fragment in a mammalian cell. In some embodiments, the promoter is inducible.

Also provided is a composition, combination or kit comprising a Cas protein and a polypeptide comprising a major coat protein (G8P) capable of binding to the Cas protein, the extracellular region of G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

In some embodiments, the polypeptides of the disclosure are provided as part of a viral particle (e.g., one whole M13 bacteriophage) or nanoparticle.

In another embodiment, the disclosure also provides a molecule comprising a Cas protein, a polypeptide comprising a major coat protein capable of binding to the Cas protein (G8P), an extracellular region of the G8P (G8P), and a cleavable linker connecting the Cas protein and the polypeptide_EX) Or the G8P or the G8P_EXA bioequivalence of (1).

In some embodiments, the cleavable linker is a peptide comprising a proteolytic cleavage site. In some embodiments, the proteolytic cleavage site is a self-cleavage site. In some embodiments, the cleavable linker is light-activated or drug-activated. In some embodiments, the Cas protein is fused to a cytidine deaminase or an adenosine deaminase.

The invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a polypeptide comprising a major coat protein capable of binding to the Cas protein (G8P), the extracellular region of G8P (G8P)_EX) Or the G8P or the G8P_EXA bioequivalence of (1). In some embodiments, the composition may be provided in the form of an injection, tablet, capsule, gel, cream, or spray.

In any embodiment, the G8P can be selected from the group consisting of SEQ ID NOs 11-20. In any embodiment, the G8P_EXSelected from the group consisting of SEQ ID NO 1-10. In any embodiment, the bioequivalence to the G8P or the G8P_EXHas at least 70% sequence identity. In any embodiment, the bioequivalent is selected from the group consisting of SEQ ID NO 37-436.

The invention also provides a polypeptide comprising an amino acid sequence derived from a sequence selected from the group consisting of SEQ ID NOs 1-22 and 37-436 comprising additions, deletions, substitutions of one, two, three, four or five amino acids, or a combination thereof, wherein the polypeptide is capable of binding to a Cas protein. Also provided is a method of genome editing in a subject, the method comprising: administering a Cas protein-based genome editing system to a subject; and administering the polypeptide or a polynucleotide encoding the polypeptide to the subject.

Drawings

Figure 1 is an in vitro activity of the major coat protein G8P from bacteriophage M13 inhibiting SpCas 9. a-b, inhibition of in vitro activity of SpCas9 by the intact M13 phage either before (a) or after (b) Cas9/sgRNA ribonucleoprotein (Cas9RNP) assembly. c-d, either before (a) or after (b) Cas9RNP assembly, by G8P_EXInhibition of in vitro activity of SpCas 9. The arrow indicates the cleavage product. Results from two biological replicates are shown as mean ± SD. The drawings are not to scale.

FIG. 2 is a diagram of determination G8P_EXAnd SpCas 9. a, alanine mutation at positions 6-11 eliminates G8P as determined by in vitro cleavage_EXInhibitory activity against SpCas 9. (a) The sequences shown in (a) include, from top to bottom: AEGDDPAKAAFDSLQASATEY (SEQ ID NO:3), mutant 1: AAAAAPAKAAFDSLQASATEY (SEQ ID NO:30), mutant 2: AEGDDAAAAAFDSLQASATEY (SEQ ID NO:31), mutant 2: AEGDDPAKAAAAAAAASATEY (SEQ ID NO:32), and mutant 4: AEGDDPAKAAFDSLQAAAAAA (SEQ ID NO: 33). b, G8P_EXStructural analysis of (2). The upper diagram shows G8P_EXResidues 6-11 are shown as rods (PDB entry 2 MJZ). This figure was generated from PyMOL. The lower panels are Wild Type (WT) and mutant 2G8P_EXCircular Dichroism (CD) spectrum of peptide. c, identification of SpCas9 and crosslinked G8P by Mass Spectrometry_EXThe interface therebetween. PyMOL shows SpCas9 structure complexed with arrestin AcrIIA4(PDBentry5VW 1). AcrIIA4 appears blue. Candidate G8P on SpCas9_EXThe binding sites are shown in green and red, respectively. Alanine scanning determined the importance of the KSVKEL (SEQ ID NO:23) and EKNPID (SEQ ID NO:34) sites for the catalytic activity of SpCas9 nuclease. The arrow indicates the cleavage product.

FIG. 3 shows G8P_EXEffect on Cas9-sgRNA binding. a, EMSA display G8P_EXDisruption of Cas9-sgRNA binding. b, G8P_EXDose-dependent inhibitory activity on Cas9-sgRNA binding. The results are shown as mean ± SD (n ═ 2). c, before or after addition of sgRNA, G8P_EXDifferential inhibition of Cas9-sgRNA binding. d, G8P_EXMutant 2 had reduced inhibitory activity on Cas9-sgRNA binding.

FIG. 4 shows G8P in mammalian cells_EXInhibition of CRISPR/Cas genome editing activity. a-b by co-delivery of G8P_EXThe peptide inhibits the genome editing activity of Cas 9/sgrnanp nuclear transfected in K562(a) and hela (b) cells. c-d by overexpression of G8P_EXInhibits genome editing activity of transiently transfected CRISPR/Cas9 in HEK293(c) and hela (d) cells. e, overexpression of G8P_EXEffect on CRISPR/Cas9 specificity. The absence or presence of G8P can be verified by Student's st test_EXSignificant difference between the specificity of CRISPR/Cas9 in case (a). f-G by over-expression of G8P_EXInhibits genome editing activity of transiently transfected CRISPR/Cas12a (Cpf1) in K562(f) and mouse N2A (g) cells. The results are shown as mean ± SD (n ═ 2 or 3). Arrows indicate the cleavage product in the T7E1 assay.

FIG. 5 shows SpCas9 and phage M13G8P_EXMass Spectrometry (MS) analysis of the interface between. In SpCas9 and G8P_EXTwo crosslinking events were identified between the peptides, including one between KSVKEL (SEQ ID NO:23) and AEGDDPAKAAF (SEQ ID NO:24) and one between ENKPIDFLEAKGY (SEQ ID NO:25) and AEGDDPAKAAF (SEQ ID NO: 24).

FIG. 6 is the construction and purification of KS and EK mutants of SpCas 9. Schematic representation of the domain organization of s. BH is a bridging helix. PI is the PAM interaction domain (KGKSKKLKSVKELLGITIMERSSFEKNPIDLEAK; SEQ ID NO: 26). Alanine mutations were located in KS (KGKSKKLAAAAAALGITIMERSSFEKNPIDLEAK; SEQ ID NO:35) and EK (KGKSKKLKSVKELLGITIMERSSFAAAAAALEAK; SEQ ID NO:36) mutants. b, WT, KS mutants and EK mutants of purified SpCas9 protein.

FIG. 7 selected study G8P_EXAn ortholog. a, selected G8P_EXListing (last column from top to bottom SEQ ID NOS: 1-10).b, in vitro shear reaction shows a different G8P_EXInhibitory activity against SpCas 9. The results are shown as mean ± SD (n ═ 2). The arrow indicates the cleavage product. 10% DMSO was used as a solvent control. G8P of c, M13 and M7_EXThe amino acid sequences of the peptides are SEQ ID NO 1 and SEQ ID NO 21, respectively. d-e, M7G 8P_EXInhibition of sgRNA binding (d) and DNA cleavage activity (e) of SpCas 9. The results are shown as mean ± SD (n ═ 2).

FIG. 8 shows G8P_EXInhibits the C-T transformation activity of A3A cytidine base editor (CBE, hAPOCC 1-nCas9-UGI) in HEK293 cells. Multiple sgrnas and target sites were analyzed by next generation sequencing. Results from two or three biological replicates are shown as mean ± SD.

FIG. 9 shows G8P_EXThe targeting specificity of A3A CBE can be improved. G8P_EXThe inhibition of treatment at the extrawindow site (C1, C12-C17) was significantly greater than at the target site (C2-C11). Multiple sgrnas and target sites were analyzed by next generation sequencing. Results from two or three biological replicates are shown as mean ± SD.

Detailed Description

Definition of

It should be noted that the term "an" object refers to one or more of the object(s); for example, "a polypeptide" is interpreted to represent one or more polypeptides. Thus, the terms "a", "an" or "the" and "at least one" may be used interchangeably herein.

Major coat proteins and fragments, and methods of use

It is herein intended to find that the major coat protein from a phage virus (i.e. G8P) can bind and prevent binding of the Cas protein to the guide nucleotide. As shown in the experimental examples, the binding can occur in the extracellular region of G8P (also referred to as "G8P_EX"), particularly between the N-terminal portion of the alpha-helix structure and a site on the Cas protein distal to the RNA or DNA binding pocket. Thus, this binding allosteric inhibits the function of Cas proteins.

Initial findings were from InoviriG8P of dae inovirus phage (M13) (including phage itself), but further experiments showed other G8P_EXFor example, G8P, produced by phages Pf1, f1, I2-2 and L.monocytogenes (Listeria) phage M7_EXBinding of Cas to RNA is also effectively inhibited. In addition, these peptides not only inhibit the function of Cas9 protein, but also bind to and have inhibitory effect on another Cas protein Cas12a (Cpf 1). These results indicate that the major coat protein is a Cas inhibitor that is widely present in bacteriophages.

The G8P protein and fragments can provide a ready solution to the off-target editing problem in genome editing. After successful editing of the target genomic site by the Cas protein-based genome editing complex, the G8P protein and fragment can prevent unwanted damage to other parts of the genome.

Thus, according to one embodiment of the present disclosure, a method of improving the specificity of a Cas protein-based genome editing process in a cell is provided. The method entails contacting the cell with the major coat protein (G8P), its extracellular portion or a biological derivative of each. The contacting can be in vitro or in vivo (e.g., in a mammalian subject).

The G8P protein and its extracellular portion (G8P) are provided in Table 1 below_EX) Have been tested for their ability to bind and inhibit Cas proteins.

Table 1 exemplary G8P and G8P_EX

As shown in example 2, G8P_EXHomolog (bioequivalence) of (1)Substances) have also been identified by informatics methods (e.g., SEQ ID NOs: 37-436) and are expected to exhibit similar binding and inhibitory activities.

As shown in example 1, G8P_EXSome of the amino acid residues in (a) are important for binding, while others are less or not. These important amino acid residues may be located, for example, in the N-terminal part of the alpha-helical structure. Thus, if G8P is modified (substituted, deleted or added)_EXOne, two or more amino acid residues in the other part, such variants are expected to retain binding and inhibitory activity. Thus, G8P and G8P_EXSuch biological equivalents of (a) are also within the scope of the present disclosure.

The term "bioequivalent" to a reference amino acid sequence refers to an amino acid sequence that has a degree of sequence identity to the reference amino acid sequence, while retaining the desired structure, function, or activity of the reference amino acid sequence. In some aspects, the sequence identity is at least about 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%. In some aspects, the bioequivalent has an addition, deletion, substitution, and combination of 1, 2, 3, 4, or 5 amino acids as compared to a reference polypeptide. G8P or G8P_EXThe desired structure of the biological equivalent of (a) is an alpha helical structure. G8P or G8P_EXThe desired activity of the bioequivalence of (a) is the ability to bind to the Cas protein and/or inhibit binding of the Cas protein to a nucleic acid molecule (e.g., sgRNA).

In some embodiments, the amino acid substitution, addition, and/or deletion is not at G8P or G8P_EXWithin the N-terminal portion of the alpha-helical structure of the sequence (e.g., the N-terminal 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid residues). In some embodiments, the amino acid substitution, addition, and/or deletion is not at G8P or G8P_EXWithin the N-terminal portion of the alpha-helical structure of the sequence (e.g., the N-terminal 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acid residues). In some embodiments, at least some or all of the amino acid substitutions, additions, and/or deletions are not in G8P or G8P_EXAlpha-helical junction of sequenceIs arranged inside the structure. In some embodiments, the G8P or G8P_EXThe bioequivalence of (a) retains the alpha-helical structure.

In some embodiments, the one or more amino acid substitutions are conservative amino acid substitutions. A "conservative amino acid substitution" is a substitution of an amino acid residue with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine tryptophan, histidine). Thus, a non-essential amino acid residue in an immunoglobulin polypeptide is preferably replaced by another amino acid residue from the same side chain family. In another embodiment, one amino acid can be replaced by another that is structurally similar, both differing in the order and/or composition of the side chain family members.

The following table provides non-limiting examples of conservative amino acid substitutions, wherein a similarity score of 0 or greater indicates that a conservative substitution has occurred between two amino acids.

TABLE A amino acid similarity matrix

	C	G	P	S	A	T	D	E	N	Q	H	K	R	V	M	I	L	F	Y	W
																					W	-8	-7	-6	-2	-6	-5	-7	-7	-4	-5	-3	-3	2	-6	-4	-5	-2	0	0	17
Y	0	-5	-5	-3	-3	-3	-4	-4	-2	-4	0	-4	-5	-2	-2	-1	-1	7	10
																					F	-4	-5	-5	-3	-4	-3	-6	-5	-4	-5	-2	-5	-4	-1	0	1	2	9
L	-6	-4	-3	-3	-2	-2	-4	-3	-3	-2	-2	-3	-3	2	4	2	6
																					I	-2	-3	-2	-1	-1	0	-2	-2	-2	-2	-2	-2	-2	4	2	5
M	-5	-3	-2	-2	-1	-1	-3	-2	0	-1	-2	0	0	2	6
																					V	-2	-1	-1	-1	0	0	-2	-2	-2	-2	-2	-2	-2	4
R	-4	-3	0	0	-2	-1	-1	-1	0	1	2	3	6
																					K	-5	-2	-1	0	-1	0	0	0	1	1	0	5
H	-3	-2	0	-1	-1	-1	1	1	2	3	6
																					Q	-5	-1	0	-1	0	-1	2	2	1	4
N	-4	0	-1	1	0	0	2	1	2
																					E	-5	0	-1	0	0	0	3	4
D	-5	1	-1	0	0	0	4
																					T	-2	0	0	1	1	3
A	-2	1	1	1	2
																					S	0	1	1	1
P	-3	-1	6
																					G	-3	5
C	12

Table B conservative amino acid substitutions

In some embodiments, G8P or G8P_EXRetains the ability to bind to the Cas protein and/or inhibit binding of the Cas protein to nucleic acid molecules such as sgrnas.

The polypeptides of the disclosure comprise G8P or G8P_EXOr bioequivalent, can be delivered as part of a fusion protein, as a separate protein, or as part of a viral particle such as a bacteriophage. As shown in the experimental examples, the intact M13 phage, the isolated G8P protein and G8P_EXThe fragments all showed inhibitory effects. Thus, in some embodiments, the polypeptide is provided by a viral particle, including a virus comprising the G8P protein or bioequivalent.

The term "Cas protein" or "Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) -associated (Cas) protein" refers to an RNA-guided DNA endonuclease that is associated with the CRISPR (clustered regularly interspaced short palindromic repeats) adaptive immune system in Streptococcus pyogenes (Streptococcus pyogenes), as well as other bacteria. Non-limiting examples of Cas proteins include Streptococcus pyogenes Cas9(SpCas9), Staphylococcus aureus (Staphylococcus aureus) Cas9(SaCas9), aminoacidococcus sp Cas12a (aspcf 1), Lachnospiraceae (Lachnospiraceae bacterium) Cas12a (LbCpf1), and Francisella novarus (Francisella noviviida) Cas12a (FnCpf 1). In Komor et al, "CRISPR-Based Technologies for the management of Eukaryotic Genomes," cell.2017Jan 12; 168(1-2): 20-36.

Non-limiting examples include SpCas9, FnCas9, St1Cas9, St3Cas9, NmCas9, SaCas9, AsCpf1, LbCpf1, FnCpf1, VQR SpCas9, EQR SpCas9, VRER SpCas9, RHA FnCas9, and KKH SaCas 9. In some embodiments, the Cas protein is a mutant of a protein selected from the group consisting of: SpCas9, FnCas9, St1Cas9, St3Cas9, NmCas9, SaCas9, AsCpf1, LbCpf1, FnCpf1, VQR SpCas9, EQR SpCas9, VRER SpCas9, RHA FnCas9 and KKH SaCas9, wherein the mutant retains DNA binding ability but does not introduce double-stranded DNA breaks. Cas proteins also include chemically modified forms that can interact with Cas protein inhibitors or chemically modified forms thereof.

For example, it is known that in SpCas9, residues Asp10 and His840 are important for the catalytic (nuclease) activity of Cas 9. When both residues were mutated to Ala, the mutant lost nuclease activity. In another embodiment, only an Asp10Ala mutation is made, and such a mutein is not capable of generating a double strand break, but rather of nicking one of the strands. This mutant is also known as Cas9 nickase.

Such and further examples are shown in table C below.

Table C exemplary Cas proteins

As used herein, "Cas protein-based genome editing process" refers to a method by which a Cas protein can be used in vitro or in vivo, in prokaryotic or eukaryotic cells, tissues, organs, or in vivo, preferably with other proteins and nucleic acids, for the purpose of genetic engineering in the genome. Examples of such methods include, but are not limited to, CRISPR-Cas gene editing, and base editing using catalytically inactive Cas (dcas) proteins, cytidine deaminase, and adenosine deaminase.

As used herein, "Cas protein-based genome editing system" refers to a composition or combination of biomolecules as desired to perform a Cas protein-based genome editing process. Such biomolecules include a Cas protein as described herein, and optionally a guide nucleic acid, a cytidine deaminase, and/or a Uracil Glycosylase Inhibitor (UGI).

In one embodiment, aFor purposes of the present disclosure, "Cas protein inhibitors" (e.g., G8P, G8P)_EXOr a biological equivalent thereof) to a cell undergoing a Cas protein-based genome editing process, e.g., genome editing or base editing. In an in vitro system, the Cas protein inhibitor may be added to a solution that includes the cells. For procedures performed on an individual, such as a patient, the Cas protein inhibitor may be administered using routes known in the art.

As observed in the examples, G8P, G8P_EXOr a biological equivalent thereof, binds to the Cas protein at a site distal to the nucleic acid binding site of the Cas protein. Thus, such binding allosterically inhibits binding of the Cas protein to a nucleic acid molecule (e.g., sgRNA). Thus, G8P, G8P are added after the Cas protein has bound to the nucleic acid_EXOr a biological equivalent thereof, does not affect the existing binding. In other words, G8P, G8P were added_EXOr a biological equivalent thereof, will only prevent or inhibit new Cas protein-nucleic acid binding. This property of these peptides has advantages.

For example, when a Cas protein-based genome editing system is introduced into a cell, initial genome editing is more likely to be correct (desired). After the initial editing stage, off-target (unwanted) editing is more likely to occur. Thus, to reduce unwanted genomic changes caused by Cas protein-based systems, G8P, G8P may be added (or induced to express) after initiation of the required editing_EXOr a biological equivalent thereof. Thus, G8P and G8P_EXOr its bioequivalence, does not affect the desired editing but only prevents/suppresses the undesired editing.

Thus, in some embodiments, a genome editing method is provided in which a Cas protein-based genome editing system and a Cas protein inhibitor (e.g., G8P, G8P) are combined_EXOr a biological equivalent thereof) to a sample (or administered to a subject). In some embodiments, the Cas protein and the Cas protein inhibitor are provided simultaneously, e.g., in the same vector-encoded conjugated composition. In such an arrangement, the Cas protein inhibitor may be operably linked to an inducible promoter, which may be at the Cas proteinThe initial genome editing phase is initiated for induction of expression.

Inducible promoters are known in the art. Chemical agents, temperature and light are examples of factors that can cause promoter induction. Common examples of chemical agents are alcohol, tetracycline, steroids, isopropyl beta-D-1-thiogalactopyranoside (IPTG), arabinose, metals, and pathogen-associated pathogens (e.g., pathogen infection. salicylic acid, ethylene, and Benzothiadiazole (BTH)).

In some embodiments, the Cas protein inhibitor is provided/administered after the Cas protein. The time of such provision/administration can be determined for a specific situation. For example, for a mammal, after administration of a Cas protein-based genome editing system, the system may require, at most, 2, 4, 6, 8, 12, 18, 24 hours or 1, 2, 3 or 4 days to initiate appropriate genome editing. Thus, the Cas protein inhibitor may be administered to the mammalian subject after this period of time. This time period can be further adjusted depending on whether the Cas protein inhibitor is administered as a protein or a polynucleotide that requires transcription and translation.

Without limitation and as described in further detail below, the Cas protein inhibitor of the present disclosure may be provided in the form of a gel, cream, solution, or aerosolized particles. Delivery to a subject may be not limited to intravenous injection, intramuscular injection, or topical administration.

Composite molecules

It is envisaged that in certain cases, Cas protein-based genome editing systems should only function at certain specified times or locations. For example, when the system is administered to a subject (human, animal, plant, etc.), it may be desirable for the system to function only in the target tissue (e.g., liver, skin). In other words, there is no need for the system to function in other tissues. The present disclosure provides a ready solution with newly discovered Cas protein inhibitors.

In one embodiment, a composite molecule is provided that includes a Cas protein, comprising a major coat protein (G8P), an extracellular region of G8P (G8P)_EX) Or G8P or G8P_EXOf (2) a bioequivalent ofA polypeptide having the ability to bind to a Cas protein (collectively Cas protein inhibitors), and a cleavable linker connecting the Cas protein and the polypeptide.

The cleavable linker can be selected or designed such that it is cleaved only in the target tissue or at the target time. For example, the cleavable linker may be photoactivatable such that the linker is cleaved only when the complexed molecule is delivered to the skin, wherein the cleavage of the linker is activated by light on the skin. In tissues without light, the complexing molecule may be in a dormant state, given that the Cas protein inhibitor binds to the Cas protein in the complexing molecule, thereby preventing it from binding to the guide nucleic acid. Once the Cas protein inhibitor is cleaved (e.g., in a skin cell), the Cas protein has an opportunity to bind to the guide nucleic acid and begin to exert its genome editing function (the cleaved Cas protein inhibitor may not have sufficient concentration to inhibit the Cas protein).

The cleavable linker may be a peptide comprising a proteolytic cleavage site. When the cleavable linker is a peptide, the composite molecule may be a fusion protein. In some embodiments, the proteolytic cleavage site comprises a self-cleaving peptide, e.g., a 2A peptide. A "2A peptide" is a viral oligopeptide of 18-22 amino acids that mediates "splicing" of polypeptides during translation in eukaryotic cells. The designation "2A" refers to a specific region of the viral genome, and different viruses 2A are typically named according to their viral origin. The first 2A discovered was F2A (foot and mouth disease virus), after which E2A (equine rhinitis virus), P2A (porcine tetanus virus-12A) and T2A (thymic virus 2A) were also identified.

As noted above, the cleavable linker may be photoactivated, or drug activated, or pH dependent, without limitation. In some embodiments, the Cas protein is fused to a cytidine deaminase or an adenosine deaminase.

Variants, fusion proteins, vectors and combinations

1-20 are wild-type sequences tested for binding and inhibitory activity of Cas proteins. Variants and homologues thereof are also expected to have such activity. For example, the bioinformatic search identified SEQ ID NO 37-436. Note that bioequivalences of each of these sequences are also within the scope of the disclosure.

In any embodiment, the Cas protein of the present disclosure can be further fused to a second protein. The second protein may be selected from a fokl nuclease domain, a transcription activator-like effector (TALE), a zinc finger protein, a transposase, a kruppel's association box (KRAB) transcription repressor, or a transcription activator.

In some embodiments, polynucleotide sequences and vectors encoding one or more of the amino acid sequences described herein are also provided. In addition to the coding sequence for the Cas protein inhibitor, the vector may also include a coding sequence for a Cas protein. Additionally or alternatively, the Cas protein or coding sequence may be provided separately in the same composition, formulation, kit or kit to facilitate simultaneous delivery. In some embodiments, the Cas protein is fused to a cytidine deaminase, e.g., A3B (APOBEC3B), A3C (APOBEC3C), A3D (APOBEC3D), A3F (APOBEC3F), A3G (APOBEC3G), A3H (APOBEC3H), a1(APOBEC1), A3(APOBEC3), and aid (aicda). In some embodiments, the Cas protein is fused to an adenosine deaminase, such as adenosine deaminase 1(Ada1) and adenosine deaminase 2(Ada2) (see, e.g., gaudell et al, Nature,551,464-71 (2017)).

In some embodiments, the Cas protein is further fused to one, two, three, or more Uracil Glycosylase Inhibitors (UGIs).

Compositions and applications

Methods of administration include, but are not limited to, intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal, epidural, and oral routes. The Cas protein inhibitor or composition may be administered by any convenient route, e.g., by infusion or bolus injection, absorbed through epithelial or cutaneous mucosal linings (e.g., oral mucosa, rectal and gastrointestinal mucosa, etc.), and may be administered with other bioactive agents. Thus, a pharmaceutical composition comprising a polypeptide of the present disclosure may be administered orally, rectally, parenterally, intrathecally, intravaginally, intraperitoneally, topically (such as by powders, ointments, drops, or transdermal patches), buccally, or in the form of an oral or nasal spray.

The term "parenteral" as used herein refers to forms of administration, which include intravenous, intramuscular, intraperitoneal, intrasternal, subcutaneous and intraarticular injection and infusion.

Administration may be systemic or local. In addition, it may be desirable to introduce the Cas protein inhibitors of the present disclosure into the central nervous system by any suitable route, including intraventricular and intrathecal injection; intraventricular injection may be facilitated by an intraventricular catheter, for example attached to a container such as an Ommaya container. Pulmonary administration can also be carried out, for example, using an inhaler or nebulizer, and formulated with an aerosolizing agent.

The Cas protein inhibitor or the composition of the present disclosure may preferably be administered topically to the area in need of treatment; this may be achieved by, for example, but not limited to, local infusion during surgery, local administration, e.g. with a wound dressing after surgery, by catheter, suppository injection, or by an implant which is a porous, non-porous or gel-like material including membranes, e.g. salivary elastic membranes or fibers. Preferably, when administering the proteins of the present disclosure, care must be taken to use materials that do not absorb the protein.

The amount of Cas protein inhibitor of the present disclosure is effective in therapy. In addition, in vitro assays may optionally be employed to help determine optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration and the severity of the disease, disorder or condition, and should be decided according to the judgment of the practitioner and each patient's circumstances. Effective doses can be extrapolated from dose response curves of in vitro or animal model test systems.

The Cas protein inhibitors of the present disclosure may be provided in the form of microparticles or nanoparticles. Thus, the preparation of microparticles or nanoparticles is also provided. The microparticles or nanoparticles may be prepared by forming an oil-in-water emulsion and then evaporating the solvent. The oily phase may be chosen from those solvents immiscible with water having a low boiling point, such as esters (e.g. ethyl acetate, butyl acetate), halogenated hydrocarbons (e.g. dichloromethane, chloroform, carbon tetrachloride, ethyl chloride, dichloroethane, trichloroethane), ethers (e.g. ethyl ether, isopropyl ether), aromatic hydrocarbons (e.g. benzene, toluene, di-ethyl ether)Toluene), carbonates (e.g., diethyl carbonate), and the like or mixtures thereof. The oil phase may also comprise various proportions of water miscible solvents (e.g., acetone) and mixtures with water immiscible solvents (e.g., dichloromethane). Suitable emulsifiers may be used to prepare the microparticles or nanoparticles to enhance the coalescence resistance of the oil droplets, wherein the emulsifiers are selected from the group including, but not limited to, polyoxyethylene sorbitan fatty acid esters such as monolaurate and trilaurate, palmitate, stearate and oleate; sorbitol fatty acid esterPolysorbatePolyvinyl alcohol, polyvinyl pyrrolidone, gelatin, lecithin, polyoxyethylene castor oil derivativesParticularly suitable is polyoxyethylene 35 castor oilAnd polyoxyethylene 40 hydrogenated castor oilA tocopherol; tocopheryl polyethylene glycol succinate (vitamin ETPGS); palmitate tocopherols and acetate tocopherols; polyoxyethylene-polyoxypropylene copolymer (C)Or) Sodium CMC, etc. or mixtures thereof. Suitable channeling agents for formulating the microparticles or nanoparticles are optionally selected from, but not limited to, polyethylene glycol, ethyl vinyl alcohol, glycerol, pentaerythritol, polyvinyl alcohol, polyvinylpyrrolidone, vinylpyrrolidone, N-methylpyrrolidone, polysaccharides such as dextrin, and/or hydrolysisStarch, sugars, sugar alcohols, and the like, or mixtures thereof.

The present disclosure also provides pharmaceutical compositions. Such compositions comprise an effective amount of protein and/or nucleotide, and an acceptable carrier. In a particular embodiment, the term "pharmaceutically acceptable" refers to drugs approved by a regulatory agency of the federal or a state government or listed in the U.S. pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans. In addition, a "pharmaceutically acceptable carrier" is generally any type of non-toxic solid, semi-solid, or liquid filler, diluent, encapsulating material, or formulation aid.

The term "carrier" refers to a diluent, adjuvant, excipient, or carrier for administration to a subject in need of treatment. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions may also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. If desired, the compositions may also contain minor amounts of wetting or emulsifying agents, or pH buffering agents such as acetates, citrates or phosphates. Antimicrobial agents such as benzyl alcohol or methylparaben, antioxidants such as ascorbic acid or sodium bisulfite, chelating agents such as ethylenediaminetetraacetic acid, and tonicity adjusting agents such as sodium chloride or dextrose are also contemplated in the present invention. These compositions may take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained release formulations and the like. The composition can be formulated into suppository with conventional binder and carrier such as triglyceride. Oral formulations may include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, and the like. Examples of suitable Pharmaceutical carriers are described in Remington's Pharmaceutical Sciences of e.w. martin, which is incorporated herein by reference. Such compositions will contain a clinically effective dose of the antigen-binding polypeptide, preferably in purified form, together with an appropriate amount of carrier to provide a form of administration suitable for the patient. The formulation should be suitable for the mode of administration. Parenteral preparations can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

In one embodiment, the composition is formulated according to conventional procedures as a pharmaceutical composition suitable for intravenous injection into a human. Typically, compositions for intravenous administration are solutions in sterile isotonic aqueous buffer. If necessary, the composition may further comprise a solubilizing agent and a local anesthetic such as lidocaine, thereby relieving pain at the injection site. In general, the active ingredients are supplied separately or mixed together in unit dosage form, e.g., as a dry lyophilized powder or as an anhydrous concentrate in a sealed container (e.g., ampoule or sachet) which is indicative of the dosage of the active ingredient. In the case of administration of the composition by infusion, the composition may be dispensed with an infusion bottle containing sterile pharmaceutical grade water or saline. In the case of administering the composition by injection, an ampoule of sterile water or saline for injection may be used so that the effective ingredients may be mixed before administration.

Examples

Example 1: inhibition of CRISPR/Cas9 by phage major coat protein-derived peptides

This example describes the following findings: the major coat protein G8P, or even the entire M13 phage, from the inovirdae inovirus phage (M13) can inhibit the activity of Streptococcus pyogenes (Streptococcus pyogenes) Cas9 nuclease (SpCas 9). Mutation analysis and high resolution mass spectrometry identified candidate interfaces between the G8P peptide and the Cas9 protein. The binding site of G8P to SpCas9 was found distal to the sgRNA or DNA binding pocket. In addition, in vitro DNA cleavage and Cas9/sgRNA gel mobility shift analysis showed that G8P inhibited Cas9 activity by preventing the formation of Cas9/sgRNA Ribonucleoprotein (RNP) complex. These results indicate that G8P is mechanistically different from previously identified anti-CRISPR proteins (Acrs). The G8P allosterically inhibits the function of SpCas 9. This example also shows that G8P orthologs from other inovirdae phages and Listeria (Listeria monocytogenes) serotype 4a (M7 strain) phages can inhibit the activity of CRISPR/Cas9, suggesting that the major coat protein is a universal mechanism for phage invasion of the bacterial immune system. This example further shows that the G8P peptide can inhibit the genome editing activity of SpCas9 in human cells.

Phylogenetic analysis showed extensive presence of Acrs in the phage. However, only a small fraction of Acrs have been experimentally validated to date. Here, it was found that phage strain M13, widely used in the laboratory, can inhibit the in vitro activity of SpCas 9. SpCas9 treatment of the intact M13 phage particles prior to assembly with the sgRNAs prevented cleavage of the substrate DNA. The inhibitory effect was dependent on the concentration of M13 phage (FIG. 1 a). Interestingly, pre-formed treatment of the Cas9/sgRNA RNP complex with phage M13 did not inhibit the cleavage reaction (fig. 1 b).

These findings facilitate the investigation of the surface protein of bacteriophage M13. This example found the extracellular region (SEQ ID NO:1) (G8P) of the major coat protein G8P (SEQ ID NO:11)_EXSEQ ID NO:1) can inhibit the activity of SpCas9 in a manner similar to that of a whole phage. Addition of G8P to Cas9 prior to RNP formation_EXCan effectively inhibit DNA shearing, and half maximum Inhibitory Concentration (IC)₅₀) About 5 μ M (FIG. 1c), whereas the activity of the pre-assembled Cas9RNP is not impaired by G8P_EXInfluence of (2) (FIG. 1 d). Importantly, the intact phage or G8P_EXDoes not impair the activity of preformed Cas9 RNPs on DNA cleavage, which indicates that phages or G8P_EXIs not a direct competitor of the sgRNA or DNA.

To understand the mechanism of inhibition, this example was performed on a 21 amino acid G8P_EXThe peptides were subjected to alanine scanning. Four peptide mutants were designed, and they were identified at G8P_EXCarry consecutive alanines on different fragments of (a). Although mutants 1, 3 and 4 showed limited or no reduced inhibitory activity against Cas9, the alanine mutations at positions 6-11 in mutant 2 abolished G8P_EXInhibitory activity of (2 a) (FIG. 2). Since this fragment contains threeThe natural alanine residue, so the other three residues P6, K8 and F11 must be aligned with G8P_EXThe inhibitory activity of (A) is related to the bond action. Structural analysis shows that G8P_EXPositions 6-11 of (a) are located at the N-terminus of the alpha helix structure. Circular Dichroism (CD) spectra show that alanine mutations in this region prevent the formation of alpha helices (fig. 2 b). G8P_EXThis alpha-helical structure may be required for interaction with Cas 9.

Next, this example attempts to examine G8P on SpCas9_EXThe binding region of (1). SpCas9 and G8P_EXCrosslinking is performed using a collision-induced dissociation (CID) cleavable crosslinker disuccinimidyl sulfoxide (DSSO). The crosslinked product was digested with chymotrypsin. MS/MS and MS by Single peptide chain fragment ions³Integrated analysis of CID-induced cleavage of cross-linked peptides revealed [ K ] from SpCas9]Cross-linked residues K1158 and E [ K ] of the SVKEL peptide]NPIDFLEAKGY (fig. 5). These peptides occupy a continuous region in the spacer Precursor Adjacent Motif (PAM) -interacting (PI) domain, which is responsible for recognition of the PAM sequence on the non-complementary DNA strand (fig. 2c and fig. 6). Interestingly, this candidate G8P_EXThe binding site is distal to the sgRNA and DNA binding pocket of Cas 9. This indicates G8P_EXIt is possible to inhibit the activity of Cas9 as an allosteric inhibitor, a mechanism different from the previously identified Acrs, which binds as a DNA mimic to the DNA binding pocket of Cas 9. These two sites were mutated to alanine, respectively, and the mutants were purified (FIG. 6). In vitro cleavage reactions showed that alanine mutations in KSKVEL (K1158 mutant) abolished cleavage activity, indicating that G8P_EXImportance of binding site to SpCas 9.

SpCas9 catalyzes activation using RNA-induced structural conformational changes. K1158 and K1176 were located on opposite surfaces of the stem-loop 1 and 2 binding domains, and thus this example analyzed G8P_EXEffect on Cas9 nuclease binding to sgRNA. Gel Electrophoresis Mobility Shift Analysis (EMSA) was performed using a fixed concentration of sgRNA and increasing concentrations of Cas9 protein. In the absence of G8P_EXIn the case of (2), coagulation was observed from Cas9 to sgRNA molar ratio of 0.1The glue is displaced. At 300. mu. M G8P_EXIn the presence, gel shifts of sgrnas binding Cas9 were observed at higher Cas9: sgrnas molar ratio, indicating perturbed interactions between Cas9 and sgrnas (fig. 3 a). G8P_EXThe inhibitory effect on Cas9-sgRNA interaction was concentration dependent. With G8P_EXAn increase in concentration formed a sgRNA complex that bound Cas9 at higher Cas9 to sgRNA molar ratios (fig. 3 b).

Similar to the shear reaction, G8P_EXInhibition of Cas9-sgRNA interaction depends on the order of sgRNA addition. With a fixed Cas9: sgRNA ratio of 0.4, Cas9 and G8P were added before sgRNA addition_EXIncubation completely inhibited formation of Cas9/sgRNA complex, at which point G8P_EXConcentrations were 300 and 600. mu.M. In contrast, when G8P was supplemented after the addition of sgRNA_EXWhen, G8P_EXComplete inhibition was not achieved at concentrations of 600 μ M or lower (FIG. 3 c). These results show that G8P_EXDoes not directly compete with sgrnas for binding to Cas 9. In addition, G8P carrying the alanine mutation at residues P6 to F11_EXMutant 2 showed inhibitory activity against the elimination of Cas9/sgRNA complex formation (fig. 3 d). This result is consistent with in vitro shearing (FIG. 2a) and provides further evidence to suggest G8P_EXNuclease activity of Cas9 was inhibited by interfering with the interaction of Cas9 with the sgRNA. Overall, the above results indicate that G8P_EXCas9 activity was inhibited by disrupting Cas9-sgRNA interaction in an allosteric format.

To study G8P_EXWhether or not the phage were a general method for inhibition of CRISPR/Cas, this example analyzed several G8P from inovirdae phage_EXPeptides (SEQ ID NOS: 2-10) (FIG. 7 a). As a result, it was found that, in addition to M13, phage G8P_EX(SEQ ID NO:1) in addition to G8P from phages Pf1, f1 and I2-2_EX(SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:9, respectively) can effectively inhibit the cleavage activity of SpCas9 in vitro (FIG. 7 b). Importantly, G8P from l.monocytogenes (listeria) (M7 strain) phage M7_EXThe orthologous peptide (SEQ ID NO:21) (FIG. 7c) effectively inhibited SpCas9 binding to the sgRNA (FIG. 7d) or cleaved the DNA substrate (FIG. 7 e). These results indicate that the main shell eggWhite G8P is a CRISPR inhibitor that is widely present in bacteriophages. For G8P from Psedomonas phage Pf1_EXMS analysis of the crosslinked Cas9 protein identified and observed M13 phage G8P_EXThe same binding site (FIG. 8), supporting G8P_EXInhibit Cas9 activity in an allosteric format.

To explore G8P_EXThis example evaluated G8P_EXEffect on genome editing activity of CRISPR/Cas in mammalian cells. This example first looks at G8P_EXWhether the cellular activity of the nuclear transfected SpCas9 protein as a co-delivered peptide could be inhibited. Analysis of T7E1 showed G8P_EXPeptides can inactivate SpCas9 protein in different genes and cell types (fig. 4 a-b). Consistent with in vitro studies, G8P was supplemented before, but not after, Cas9/sgRNA RNP complex formation_EXPeptide, the genome editing activity of SpCas9 protein was abolished (fig. 4 a-b). These data further support G8P_EXAs an indirect competitor of sgRNA or DNA to inhibit the activity of Cas 9. Next, this example evaluated over-expressed G8P_EXEffect on transiently transfected CRISPR/Cas 9. Transient transfection of Cas9 and sgRNA-encoding plasmids into human cells by lipofection, 6h post-transfection G8P_EXAn overexpression plasmid. G8P was found in various genes and cell types_EXInhibition of the genome editing activity of transiently transfected SpCas9 by 50% or more (fig. 4 c-d). Importantly, as in the absence or presence of G8P_EXAs confirmed by the on/off shear activity in the case of (1), G8P_EXThe activity of Cas9 was inhibited without compromising its specificity (fig. 4 e). To study G8P_EXWhether it is a universal switch for different CRISPR/Cas systems, this example investigates its inhibitory effect on CRISPR/Cas12a (Cpf 1). It was found that G8P_EXThe genome editing activity of Cpf1 was inhibited in human and mouse cells (fig. 4 f-g).

In addition to inhibiting the cleavage activity of DNA of CRISPR/Cas9, G8P_EXCan also be used to modulate the activity of Cas 9-derived base editors. We show G8P_EXCan inhibit C-T transition induced by A3A cytidine base editor (CBE, hAPOCC 1-nCas9-UGI) in HEK293 cellsAnd (4) transforming. This inhibitory activity was observed at multiple genomic sites of different sgrnas (fig. 8). Furthermore, in the same target site, G8P_EXDifferent degrees of inhibition were shown at different cytidine sites.

A3A CBE can induce C-T transformation at the target site (on-target site, positions 2-11) among 20-bp target sites. In addition, A3A CBE can induce C-T transformation at the out-of-window sites (positions 1 and 12-20). Surprisingly, we found that G8P_EXA significantly stronger inhibition was shown at sites outside the window than at the target site (fig. 9). In other words, G8P_EXWhere present, the ratio between the target site and the site outside the window is increased. This indicates G8P_EXCan be used to improve target specificity at a target site.

Example 2 search G8P_EXHomologous sequences

G8P to be successfully tested in example 1_EXThe sequence was used as input sequence for searching for homologues. Using NCBI BLAST^TMProgram, and the following parameters (Expect threshold:10, Word size:6, Max matches in a query range:0, Matrix: blosum62, Gap codes: Existence:11Extension:1, Compositional adjustments: Conditional Compositional Matrix adjustment).

As shown in table 2 below, 400 important shoot head sequences were identified. These sequences and variants thereof are also expected to have the ability to bind Cas proteins and inhibit their function of binding nucleic acids.

TABLE 2G8P_EXHomologues

***

The scope of the invention is not limited by the specific examples described, which are intended as illustrations of individual aspects, and any compositions or methods that are functionally equivalent are within the scope of this disclosure. It will be apparent to those skilled in the art that various modifications and variations can be made in the methods and compositions of the present invention without departing from the spirit or scope of the invention. Thus, it is intended that the present disclosure cover the modifications and variations of this invention provided they come within the scope of the appended claims and their equivalents.

All publications and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.

SEQUENCE LISTING

<110> Shanghai science and technology university

Composition of <120> Cas protein inhibitor and application

<130> P21112018CP

<150> PCT/CN2018/102908

<151> 2018-08-29

<160> 436

<170> PatentIn version 3.5

<210> 1

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 1

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

1 5 10 15

Ser Ala Thr Glu Tyr

20

<210> 2

<211> 29

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 2

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

1 5 10 15

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

20 25

<210> 3

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 3

Ala Glu Gly Asp Asp Pro Ala Lys Ala Ala Phe Asp Ser Leu Gln Ala

1 5 10 15

Ser Ala Thr Glu Tyr

20

<210> 4

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 4

Met Gln Ser Val Ile Thr Asp Val Thr Gly Gln Leu Thr Ala Val Gln

1 5 10 15

Ala Asp

<210> 5

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 5

Ala Glu Pro Asn Ala Ala Thr Asn Tyr Ala Thr Glu Ala Met Asp Ser

1 5 10 15

Leu Lys Thr Gln Ala Ile Asp Leu Ile

20 25

<210> 6

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 6

Ala Asp Asp Ala Thr Ser Gln Ala Lys Ala Ala Phe Asp Ser Leu Thr

1 5 10 15

Ala Gln Ala Thr Glu Met

20

<210> 7

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 7

Ser Gly Val Gly Asp Gly Val Asp Val Val Ser Ala Ile Glu Gly Ala

1 5 10 15

Ala Gly Pro

<210> 8

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 8

Met Asp Phe Asn Pro Ser Glu Val Ala Ser Gln Val Thr Asn Tyr Ile

1 5 10 15

Gln

<210> 9

<211> 29

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 9

Ala Asp Asp Gly Thr Ser Thr Ala Thr Ser Tyr Ala Thr Glu Ala Met

1 5 10 15

Asn Ser Leu Lys Thr Gln Ala Thr Asp Leu Ile Asp Gln

20 25

<210> 10

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 10

Met Gly Asp Ile Leu Thr Gly Val Ser Gly Ala Glu

1 5 10

<210> 11

<211> 73

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 11

Met Lys Lys Ser Leu Val Leu Lys Ala Ser Val Ala Val Ala Thr Leu

1 5 10 15

Val Pro Met Leu Ser Phe Ala Ala Glu Gly Asp Asp Pro Ala Lys Ala

20 25 30

Ala Phe Asn Ser Leu Gln Ala Ser Ala Thr Glu Tyr Ile Gly Tyr Ala

35 40 45

Trp Ala Met Val Val Val Ile Val Gly Ala Thr Ile Gly Ile Lys Leu

50 55 60

Phe Lys Lys Phe Thr Ser Lys Ala Ser

65 70

<210> 12

<211> 82

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 12

Met Lys Ala Met Lys Gln Arg Ile Ala Lys Phe Ser Pro Val Ala Ser

1 5 10 15

Phe Arg Asn Leu Cys Ile Ala Gly Ser Val Thr Ala Ala Thr Ser Leu

20 25 30

Pro Ala Phe Ala Gly Val Ile Asp Thr Ser Ala Val Glu Ser Ala Ile

35 40 45

Thr Asp Gly Gln Gly Asp Met Lys Ala Ile Gly Gly Tyr Ile Val Gly

50 55 60

Ala Leu Val Ile Leu Ala Val Ala Gly Leu Ile Tyr Ser Met Leu Arg

65 70 75 80

Lys Ala

<210> 13

<211> 73

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 13

Met Lys Lys Ser Leu Val Leu Lys Ala Ser Val Ala Val Ala Thr Leu

1 5 10 15

Val Pro Met Leu Ser Phe Ala Ala Glu Gly Asp Asp Pro Ala Lys Ala

20 25 30

Ala Phe Asp Ser Leu Gln Ala Ser Ala Thr Glu Tyr Ile Gly Tyr Ala

35 40 45

Trp Ala Met Val Val Val Ile Val Gly Ala Thr Ile Gly Ile Lys Leu

50 55 60

Phe Lys Lys Phe Thr Ser Lys Ala Ser

65 70

<210> 14

<211> 44

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 14

Met Gln Ser Val Ile Thr Asp Val Thr Gly Gln Leu Thr Ala Val Gln

1 5 10 15

Ala Asp Ile Thr Thr Ile Gly Gly Ala Ile Ile Val Leu Ala Ala Val

20 25 30

Val Leu Gly Ile Arg Trp Ile Lys Ala Gln Phe Phe

35 40

<210> 15

<211> 82

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 15

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

1 5 10 15

Gly Ala Ala Thr Met Leu Val Ser Ala Gly Ser Phe Ala Ala Glu Pro

20 25 30

Asn Ala Ala Thr Asn Tyr Ala Thr Glu Ala Met Asp Ser Leu Lys Thr

35 40 45

Gln Ala Ile Asp Leu Ile Ser Gln Thr Trp Pro Val Val Thr Thr Val

50 55 60

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

65 70 75 80

Ala Val

<210> 16

<211> 74

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 16

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

1 5 10 15

Ile Gly Ser Ser Ala Phe Ala Ala Asp Asp Ala Thr Ser Gln Ala Lys

20 25 30

Ala Ala Phe Asp Ser Leu Thr Ala Gln Ala Thr Glu Met Ser Gly Tyr

35 40 45

Ala Trp Ala Leu Val Val Leu Val Val Gly Ala Thr Val Gly Ile Lys

50 55 60

Leu Phe Lys Lys Phe Val Ser Arg Ala Ser

65 70

<210> 17

<211> 44

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 17

Ser Gly Val Gly Asp Gly Val Asp Val Val Ser Ala Ile Glu Gly Ala

1 5 10 15

Ala Gly Pro Ile Ala Ala Ile Gly Gly Ala Val Leu Thr Val Met Val

20 25 30

Gly Ile Lys Val Tyr Lys Trp Val Arg Arg Ala Met

35 40

<210> 18

<211> 46

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 18

Met Asp Phe Asn Pro Ser Glu Val Ala Ser Gln Val Thr Asn Tyr Ile

1 5 10 15

Gln Ala Ile Ala Ala Ala Gly Val Gly Val Leu Ala Leu Ala Ile Gly

20 25 30

Leu Ser Ala Ala Trp Lys Tyr Ala Lys Arg Phe Leu Lys Gly

35 40 45

<210> 19

<211> 84

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 19

Met Ser Val Ile Thr Lys Val Ala Ala Ala Lys Asn Lys Ile Val Val

1 5 10 15

Gly Ala Gly Leu Leu Met Ala Ser Ala Gly Ala Phe Ala Ala Asp Asp

20 25 30

Gly Thr Ser Thr Ala Thr Ser Tyr Ala Thr Glu Ala Met Asn Ser Leu

35 40 45

Lys Thr Gln Ala Thr Asp Leu Ile Asp Gln Thr Trp Pro Val Val Thr

50 55 60

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

65 70 75 80

Ser Lys Ala Val

<210> 20

<211> 42

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 20

Met Gly Asp Ile Leu Thr Gly Val Ser Gly Ala Glu Ala Ala Thr Ala

1 5 10 15

Met Ile Ala Ala Ala Ala Ile Ile Ala Leu Val Gly Phe Thr Lys Trp

20 25 30

Gly Ala Lys Lys Val Ala Ser Phe Phe Gly

35 40

<210> 21

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 21

Ser Lys Gly Asn Asp Gln Val Phe Thr Arg Arg Ala Phe Thr Met His

1 5 10 15

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

20 25 30

Thr Pro Thr Asn Ala Asp Leu

35

<210> 22

<211> 333

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 22

Met Ala Asn Glu Ile Thr Lys Leu Leu Asp Val Val Thr Pro Glu Val

1 5 10 15

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

20 25 30

Gln Ser Gly Ile Ala Val Ala Asp Pro Ser Val Ala Gln Asn Ile Thr

35 40 45

Ala Gly Gly Leu Leu Val Asn Met Pro Phe Trp Asn Asp Leu Asp Gly

50 55 60

Glu Asp Glu Thr Leu Gly Asp Gly Glu Lys Gly Leu Glu Thr Gly Lys

65 70 75 80

Ile Thr Ala Ser Ala Asp Ile Ala Ala Val Met Tyr Arg Gly Arg Gly

85 90 95

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

100 105 110

Asp Ala Leu Met Gly Lys Ile Ala Ser Trp Trp Met Arg Arg Glu Gln

115 120 125

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

130 135 140

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

145 150 155 160

Asn Leu Val Leu Asp Ala Lys Gln Leu Leu Gly Asp Ser Ser Asp Arg

165 170 175

Leu Ser Leu Met Val Met His Ser Ala Val Tyr Thr Ala Leu Gln Lys

180 185 190

Gln Asn Leu Ile Ala Phe Ile Pro Asn Ala Arg Gly Glu Val Asn Ile

195 200 205

Pro Thr Tyr Leu Gly Tyr Arg Val Val Val Asp Asp Gly Val Pro Ser

210 215 220

Thr Gly Thr Gly Ala Ala Lys Val Tyr Thr Ser Tyr Leu Phe Ala Thr

225 230 235 240

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

245 250 255

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

260 265 270

Arg Ala Phe Thr Met His Pro Tyr Gly Val Lys Phe Lys Asn Ala Val

275 280 285

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

290 295 300

Gly Asn Trp Glu Lys Val Tyr Glu Asp Lys Gln Ile Gly Ile Val Gly

305 310 315 320

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

325 330

<210> 23

<211> 6

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 23

Lys Ser Val Lys Glu Leu

1 5

<210> 24

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 24

Ala Glu Gly Asp Asp Pro Ala Lys Ala Ala Phe

1 5 10

<210> 25

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 25

Glu Asn Lys Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr

1 5 10

<210> 26

<211> 34

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 26

Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile

1 5 10 15

Thr Ile Met Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Leu Glu

20 25 30

Ala Lys

<210> 27

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 27

Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr

1 5 10

<210> 28

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 28

Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr

1 5 10

<210> 29

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 29

Val Arg Gln Gln Leu Pro Glu Lys Tyr

1 5

<210> 30

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 30

Ala Ala Ala Ala Ala Pro Ala Lys Ala Ala Phe Asp Ser Leu Gln Ala

1 5 10 15

Ser Ala Thr Glu Tyr

20

<210> 31

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 31

Ala Glu Gly Asp Asp Ala Ala Ala Ala Ala Phe Asp Ser Leu Gln Ala

1 5 10 15

Ser Ala Thr Glu Tyr

20

<210> 32

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 32

Ala Glu Gly Asp Asp Pro Ala Lys Ala Ala Ala Ala Ala Ala Ala Ala

1 5 10 15

Ser Ala Thr Glu Tyr

20

<210> 33

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 33

Ala Glu Gly Asp Asp Pro Ala Lys Ala Ala Phe Asp Ser Leu Gln Ala

1 5 10 15

Ala Ala Ala Ala Ala

20

<210> 34

<211> 6

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 34

Glu Lys Asn Pro Ile Asp

1 5

<210> 35

<211> 34

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 35

Lys Gly Lys Ser Lys Lys Leu Ala Ala Ala Ala Ala Ala Leu Gly Ile

1 5 10 15

Thr Ile Met Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Leu Glu

20 25 30

Ala Lys

<210> 36

<211> 34

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 36

Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile

1 5 10 15

Thr Ile Met Glu Arg Ser Ser Phe Ala Ala Ala Ala Ala Ala Leu Glu

20 25 30

Ala Lys

<210> 37

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 37

Gly Thr Ala Thr Ala Thr Ser

1 5

<210> 38

<211> 7

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 38

Met Asp Phe Asp Pro Ser Glu

1 5

<210> 39

<211> 8

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 39

Met Asp Phe Asn Pro Ser Glu Val

1 5

<210> 40

<211> 8

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 40

Met Glu Phe Asn Pro Ser Asp Val

1 5

<210> 41

<211> 8

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 41

Pro Ser Glu Val Ala Ser Gln Val

1 5

<210> 42

<211> 8

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 42

Ser Gln Val Thr Asn Tyr Ile Gln

1 5

<210> 43

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 43

Ala Ser Gln Val Thr Asp Tyr Ile Gln

1 5

<210> 44

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 44

Asp Phe Asn Pro Ser Ala Val Ala Ser

1 5

<210> 45

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 45

Phe Asn Pro Ser Asp Val Ala Pro Gln

1 5

<210> 46

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 46

Phe Asp Pro Ser Glu Val Ala Pro Gln

1 5

<210> 47

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 47

Met Asp Phe Asp Pro Ser Glu Val Ala

1 5

<210> 48

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 48

Met Asp Phe Glu Pro Ser Glu Val Ala

1 5

<210> 49

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 49

Asn Pro Ala Glu Val Ala Ser Gln Val

1 5

<210> 50

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 50

Pro Ser Glu Val Ala Ser Gln Val Thr

1 5

<210> 51

<211> 9

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 51

Thr Gln Ala Thr Asp Leu Leu Asp Gln

1 5

<210> 52

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 52

Asp Phe Asn Pro Thr Asp Val Ala Ser Gln

1 5 10

<210> 53

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 53

Asp Phe Asn Tyr Pro Ser Glu Val Ala Ser

1 5 10

<210> 54

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 54

Asp Thr Ser Ala Ile Glu Ala Ala Ile Thr

1 5 10

<210> 55

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 55

Glu Ala Ala Ser Gln Val Thr Asn Tyr Ile

1 5 10

<210> 56

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 56

Met Asp Ile Asn Pro Ser Asp Ile Ala Ser

1 5 10

<210> 57

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 57

Met Asn Ala Leu Lys Thr Thr Asp Leu Ile

1 5 10

<210> 58

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 58

Asn Pro Ser Glu Val Ala Pro Gln Val Thr

1 5 10

<210> 59

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 59

Pro Ala Phe Ala Gly Val Ile Asp Thr Ser

1 5 10

<210> 60

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 60

Ser Ala Ala Ser Ser Tyr Ala Ala Glu Ala

1 5 10

<210> 61

<211> 10

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 61

Ser Ala Ala Asp Ser Tyr Asp Ala Asp Ala

1 5 10

<210> 62

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 62

Ala Leu Thr Glu Ala Gln Gly Asp Met Lys Ala

1 5 10

<210> 63

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 63

Asp Asp Gly Thr Ala Thr Ala Thr Ser Tyr Ala

1 5 10

<210> 64

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 64

Asp Ser Leu Lys Thr Gln Ala Ile Asp Leu Ile

1 5 10

<210> 65

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 65

Glu Asp Glu Ala Lys Ala Ala Phe Glu Ala Leu

1 5 10

<210> 66

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 66

Glu Val Ala Ala Gln Val Thr Asp Tyr Leu Gln

1 5 10

<210> 67

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 67

Glu Val Val Ser Asp Leu Thr Asn Tyr Ile Gln

1 5 10

<210> 68

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 68

Phe Asn Pro Ser Glu Ile Pro Ser Gln Ile Thr

1 5 10

<210> 69

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 69

Gly Thr Pro Thr Ala Thr Ser Tyr Ala Thr Glu

1 5 10

<210> 70

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 70

Gly Thr Ser Ala Ala Thr Ser Tyr Asp Thr Glu

1 5 10

<210> 71

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 71

Met Asp Phe Asp Pro Ser Glu Leu Ala Tyr Ile

1 5 10

<210> 72

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 72

Met Asp Phe Asn Pro Glu Met Ala Ser Ser Val

1 5 10

<210> 73

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 73

Met Asp Phe Asn Pro Ser Asp Ile Ala Tyr Ile

1 5 10

<210> 74

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 74

Met Asn Ser Leu Lys Asn His Thr Asp Leu Ile

1 5 10

<210> 75

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 75

Met Ser Phe Asn Pro Glu Val Ala Ser Gln Val

1 5 10

<210> 76

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 76

Asn Asp Gly Thr Ser Thr Ala Thr Ser Ala Thr

1 5 10

<210> 77

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 77

Asn Phe Asn Pro Gln Val Thr Asn Tyr Ile Glu

1 5 10

<210> 78

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 78

Asn Pro Ser Glu Val Ala Ser Gln Ile Ile Gln

1 5 10

<210> 79

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 79

Pro Ser Ala Val Gln Val Thr Asn Tyr Ile Gln

1 5 10

<210> 80

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 80

Ser Glu Val Gly Gly Asn Met Thr Ser Tyr Ile

1 5 10

<210> 81

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 81

Ser Gln Val Ala Ser Gln Val Thr Asp Tyr Ile

1 5 10

<210> 82

<211> 11

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 82

Thr Glu Ile Met Gly Ser Leu Lys Thr Gln Ala

1 5 10

<210> 83

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 83

Ala Ala Asp Ser Leu Gln Ala Ser Ala Thr Asp Tyr

1 5 10

<210> 84

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 84

Ala Met Asn Ser Leu Lys Glu Asn Asp Leu Ile Asp

1 5 10

<210> 85

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 85

Asp Asp Ala Ser Ser Asn Ala Thr Ser Tyr Ala Thr

1 5 10

<210> 86

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 86

Asp Asp Gly Thr Ala Asn Ala Thr Ser Tyr Ala Thr

1 5 10

<210> 87

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 87

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

1 5 10

<210> 88

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 88

Asp Phe Val Ala Ser Gln Val Thr Asn Tyr Lys Gln

1 5 10

<210> 89

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 89

Glu Val Ala Ser Gln Leu Leu Gly Thr Asn Tyr Ile

1 5 10

<210> 90

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 90

Phe Asn Pro Pro Thr Val Thr Ser Gln Val Thr Asn

1 5 10

<210> 91

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 91

Gly Thr Ser Thr Thr Thr Thr Tyr Thr Asp Ala Met

1 5 10

<210> 92

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 92

Ile Asp Thr Ser Leu Ile Glu Ala Ala Ile Thr Asp

1 5 10

<210> 93

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 93

Leu Asp Leu Ser Pro Ser Glu Val Ala Ser Gln Val

1 5 10

<210> 94

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 94

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

1 5 10

<210> 95

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 95

Met Asp Glu Ile Ser Gln Ile Thr Asp Tyr Ile Gln

1 5 10

<210> 96

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 96

Met Asp Phe Thr Pro Ser Glu Val Ala Arg Asn Val

1 5 10

<210> 97

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 97

Asn Pro Asn Glu Val Ser Arg Gln Val Asn Tyr Ile

1 5 10

<210> 98

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 98

Asn Pro Arg Glu Ile Ala Ser Gln Val Ser Asp Tyr

1 5 10

<210> 99

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 99

Asn Pro Ser Glu Val Ala Ser Gln Ile Ile Ile Gln

1 5 10

<210> 100

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 100

Asn Ser Ser Glu Val Ala Ser Gln Val Ala Asn Tyr

1 5 10

<210> 101

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 101

Pro Ala Lys Ala Ala Phe Asp Leu Gln Ala Thr Ala

1 5 10

<210> 102

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 102

Ser Glu Val Ala Ser Gln Val Gln Asn Phe Ile Gln

1 5 10

<210> 103

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 103

Ser Phe Ala Thr Glu Val Met Asn Ser Leu Pro Thr

1 5 10

<210> 104

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 104

Ser Tyr Ala Thr Ser Met Asn Asn Leu Lys Lys Gln

1 5 10

<210> 105

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 105

Thr Ala Ser Ser Tyr Ala Thr Gln Ala Gln Lys Ser

1 5 10

<210> 106

<211> 12

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 106

Met Gly Asp Ile Leu Thr Gly Val Ser Gly Ala Glu

1 5 10

<210> 107

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 107

Ala Gly Leu Ile Asp Thr Ala Ser Val Glu Ser Asp Ile

1 5 10

<210> 108

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 108

Ala Gly Leu Ile Asp Thr Ser Ser Val Asp Asn Ala Ile

1 5 10

<210> 109

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 109

Ala Gly Leu Ile Asp Thr Ser Ser Val Glu Lys Ser Ile

1 5 10

<210> 110

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 110

Ala Gly Leu Ile Glu Thr Ser Ser Val Glu Arg Ser Ile

1 5 10

<210> 111

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 111

Ala Gly Leu Ile Asp Thr Ser Ser Val Glu Ser Ser Ile

1 5 10

<210> 112

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 112

Ala Gly Leu Ile Asp Thr Ser Thr Val Asp Ser Thr Ile

1 5 10

<210> 113

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 113

Ala Gly Met Ile Asp Ser Ser Ser Val Asp Ser Ala Ile

1 5 10

<210> 114

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 114

Ala Gly Met Ile Asp Thr Ser Ser Ile Asp Ser Ser Ile

1 5 10

<210> 115

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 115

Asp Asp Gly Thr Thr Thr Ala Tyr Ala Thr Asp Glu Ala

1 5 10

<210> 116

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 116

Asp Phe Asp Pro Ser Glu Val Glu Gln Val Val Asn Tyr

1 5 10

<210> 117

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 117

Asp Phe Asn Pro Ala Glu Val Ala Lys Arg Asn Tyr Ile

1 5 10

<210> 118

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 118

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

1 5 10

<210> 119

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 119

Asp Phe Ser Pro Ala Glu Val Ala Lys Arg Asn Tyr Ile

1 5 10

<210> 120

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 120

Asp Phe Ser Gln Ser Glu Val Asp Gln Gln Val Thr Asn

1 5 10

<210> 121

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 121

Asp Ser Gly Thr Ser Ala Ala Thr Ser Tyr Asp Thr Glu

1 5 10

<210> 122

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 122

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

1 5 10

<210> 123

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 123

Phe Asn Pro Asn Glu Val Leu Ala Ser Gln Ser Thr Asn

1 5 10

<210> 124

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 124

Phe Asn Pro Ser Glu Val Ala Ala Leu Thr Ser Phe Ile

1 5 10

<210> 125

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 125

Phe Asn Pro Ser Glu Val Pro Ser Ile Gly Asn Tyr Ile

1 5 10

<210> 126

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 126

Met Asp Asp Phe Asn Pro Ala Glu Val Ala Arg Gln Ile

1 5 10

<210> 127

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 127

Met Asp Phe Tyr Gln Lys Asn Pro Ser Ala Val Ala Ser

1 5 10

<210> 128

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 128

Met Ser Phe Asn Ser Glu Ser Ala Ser Gln Val Thr Asn

1 5 10

<210> 129

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 129

Met Ser Phe Ser Pro Ser Glu Ile Ala Ser Glu Ile Gln

1 5 10

<210> 130

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 130

Pro Ser Glu Val Ala Ala Gln Ile Val Asn Tyr Tyr Gln

1 5 10

<210> 131

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 131

Ser Ala Val Glu Ser Ala Phe Asn Glu Gly Glu Gly Asp

1 5 10

<210> 132

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 132

Thr Ser Tyr Ala Thr Asn Ser Ile Asn Ser Leu Asn Thr

1 5 10

<210> 133

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 133

Thr Ser Tyr Ala Thr Ser Ser Ile Asn Ser Leu Asn Thr

1 5 10

<210> 134

<211> 13

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 134

Val Leu Asp Thr Ser Glu Val Glu Ser Pro Ile Thr Glu

1 5 10

<210> 135

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 135

Ala Asp Glu Gly Thr Thr Ala Thr Thr Tyr Ala Val Asp Ala

1 5 10

<210> 136

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 136

Ala Glu Asp Gly Thr Thr Ala Thr Thr Tyr Ala Val Glu Ala

1 5 10

<210> 137

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 137

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

1 5 10

<210> 138

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 138

Ala Phe Cys Gly Val Ile Asp Thr Ser Ala Val Ala Ser Ala

1 5 10

<210> 139

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 139

Ala Gly Asp Gly Thr Thr Thr Ala Thr Ile Tyr Ala Glu Ala

1 5 10

<210> 140

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 140

Ala Gly Glu Gly Thr Ser Thr Ala Thr Val Tyr Ala Glu Ala

1 5 10

<210> 141

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 141

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

1 5 10

<210> 142

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 142

Ala Ser Val Ala Asp Ser Ser Ala Ala Glu Ser Ala Ile Thr

1 5 10

<210> 143

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 143

Asp Met Thr Ala Val Asn Ser Ala Ile Thr Ala Gly Gln Gly

1 5 10

<210> 144

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 144

Phe Asn Pro Thr Glu Val Val Gln Val Thr Gln Tyr Leu Gln

1 5 10

<210> 145

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 145

Phe Ser Pro Glu Val Ile Ser Gln Ile Thr Glu Tyr Ile Gln

1 5 10

<210> 146

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 146

Gly Gln Ala Thr Ala Thr Ser Tyr Ala Thr Gln Thr Gln Ala

1 5 10

<210> 147

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 147

Gly Val Ile Asp Ala Ser Ala Val Glu Arg Ala Ile Ala Asp

1 5 10

<210> 148

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 148

Leu Asn Phe Glu Ile Ser Glu Val Ala Ser Gln Val Thr Asn

1 5 10

<210> 149

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 149

Met Asp Phe Asn Arg Ser Gly Val Thr Ser Ser Val Thr Asn

1 5 10

<210> 150

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 150

Met Asp Ile Asn Ser Ser Glu Val Ala Thr Asp Phe Tyr Ile

1 5 10

<210> 151

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 151

Met Gly Phe Asn Pro Ser Ile Asp Val Arg Gln Val Thr Asn

1 5 10

<210> 152

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 152

Asn Pro Asn Glu Val Ala Glu His Val Thr Ala Tyr Ile Glu

1 5 10

<210> 153

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 153

Asn Pro Ser Asp Ile Ala Ser Gln Val Ala Asp Tyr Phe Gln

1 5 10

<210> 154

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 154

Pro Ser Glu Ile Ala Asn Gln Ile Thr Asn Tyr Leu Ile Gln

1 5 10

<210> 155

<211> 14

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 155

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

1 5 10

<210> 156

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 156

Ala Gly Val Ile Asp Arg Ser Ala Leu Gln Ser Ala Met Lys Ala

1 5 10 15

<210> 157

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 157

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

1 5 10 15

<210> 158

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 158

Ala Lys Glu Ala Ile Asn Ser Leu Lys Thr Gln Ser Lys Asp Leu

1 5 10 15

<210> 159

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 159

Ala Met Lys Ser Ile Leu Lys Thr Thr Ala Asp Leu Val Asp Gln

1 5 10 15

<210> 160

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 160

Ala Thr Glu Ala Met Asp Ser Leu Glu Thr Gln Thr Ala Thr Asp

1 5 10 15

<210> 161

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 161

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

1 5 10 15

<210> 162

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 162

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

1 5 10 15

<210> 163

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 163

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

1 5 10 15

<210> 164

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 164

Asp Ala Ala Lys Ala Ala Tyr Asp Ser Leu Arg Ala Gly Ala Thr

1 5 10 15

<210> 165

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 165

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

1 5 10 15

<210> 166

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 166

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

1 5 10 15

<210> 167

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 167

Asp Gly Thr Ser Thr Ala Thr Ala Thr Ser Ser Glu Ala Ala Asn

1 5 10 15

<210> 168

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 168

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

1 5 10 15

<210> 169

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 169

Glu Ala Met Asn Lys Leu Lys Ala Gln Ala Lys Glu Val Ile Asp

1 5 10 15

<210> 170

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 170

Glu Ala Met Asn Val Leu Ser Ile Leu Glu Thr Asp Leu Ile Asp

1 5 10 15

<210> 171

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 171

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

1 5 10 15

<210> 172

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 172

Glu Ala Met Thr Asn Leu Asp Thr Gln Ala Ile Asn Ala Ile Asp

1 5 10 15

<210> 173

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 173

Glu Gly Ala Asp Pro Ala Ala Val Ala Phe Asp Ser Leu Gln Ala

1 5 10 15

<210> 174

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 174

Asp Gly Ser Glu Pro Ala Ala Val Ala Phe Asp Ala Leu Gln Ala

1 5 10 15

<210> 175

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 175

Glu Gly Ser Asp Pro Ala Ala Val Ala Phe Glu Thr Leu Gln Ala

1 5 10 15

<210> 176

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 176

Phe Asn Pro Ser Glu Thr Ile Ser Gln Leu Thr His Tyr Ile Glu

1 5 10 15

<210> 177

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 177

Leu Pro Ala Phe Leu Gly Ile Ile Ala Thr Ser Thr Val Ser Ala

1 5 10 15

<210> 178

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 178

Met Gly Phe Asn Pro Phe Val Ser Glu Val Ala His Gln Leu Thr

1 5 10 15

<210> 179

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 179

Met His Phe Asn Val Ser Asp Ala Ala Asn Gln Ile Thr Asn Tyr

1 5 10 15

<210> 180

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 180

Asn Pro Glu Val Ala Asp Ser Ser Gln Cys Thr Ser Tyr Ile Gln

1 5 10 15

<210> 181

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 181

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

1 5 10 15

<210> 182

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 182

Gln Ala Met Asn Glu Ala Leu Lys Thr Ser Ala Thr Asp Leu Ile

1 5 10 15

<210> 183

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 183

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

1 5 10 15

<210> 184

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 184

Thr Ala Val Ser Ile Ala Thr Glu Ala Met Asp Arg Leu Arg Thr

1 5 10 15

<210> 185

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 185

Thr Glu Ile Met Asn Ser Leu Lys Asn Ser Asp Thr Asp Leu Ile

1 5 10 15

<210> 186

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 186

Thr Ser Leu Pro Ala Phe Ala Ala Leu Ile Asp Ser Ala Ala Val

1 5 10 15

<210> 187

<211> 15

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 187

Thr Ser Thr Ala Thr Ser Tyr Ser Thr Glu Leu Arg Thr Ala Met

1 5 10 15

<210> 188

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 188

Ala Thr Ala Leu Pro Ala Phe Ala Thr Ser Ala Val Ala Ala Ile Thr

1 5 10 15

<210> 189

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 189

Ala Thr Ser Phe Ser Thr Glu Ala Val Asn Ser Leu Thr Asn Leu Ile

1 5 10 15

<210> 190

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 190

Ala Thr Ser Tyr Ala Thr Ala Glu Pro Met Asp Ala Thr Gln Ala Thr

1 5 10 15

<210> 191

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 191

Asp Phe Leu Asn Pro Ser Glu Val Ala Ser Leu Gln Gln Val Tyr Asn

1 5 10 15

<210> 192

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 192

Asp Phe Asn Pro Ser Ala Ser Ala Asp Val Ala Ala Ala Ile Thr Asn

1 5 10 15

<210> 193

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 193

Asp Gly Thr Ser Ser Thr Ser Tyr Glu Thr Val Pro Leu Asn Ser Leu

1 5 10 15

<210> 194

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 194

Glu Ala Met Asn Ser Leu Thr Thr Gln Ala Lys Ala Asp Leu Leu Glu

1 5 10 15

<210> 195

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 195

Gly Ile Ile Asp Thr Asn Val Val Gln Ser Ala Ile Ser Asp Ala Gln

1 5 10 15

<210> 196

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 196

Gly Val Ile Arg Asp Thr Pro Glu Ala Val Glu Ser Ala Phe Thr Asp

1 5 10 15

<210> 197

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 197

Met Asp Phe Asn Pro Ser Glu Ile Ala Thr Thr Val Val Gln Tyr Ile

1 5 10 15

<210> 198

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 198

Met Glu Phe His Ala Asp Glu Val Ala Ala Asn Val Thr Gly Tyr Ile

1 5 10 15

<210> 199

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 199

Met Gly Phe Asn Pro Arg Glu Val Ser Lys Gln Val Ile Glu Asn Tyr

1 5 10 15

<210> 200

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 200

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

1 5 10 15

<210> 201

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 201

Asn Phe Asn Ala Ser Glu Pro Ser Gln Leu Val Thr Lys Tyr Ile Gln

1 5 10 15

<210> 202

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 202

Thr Ala Ile Ser Tyr Gln Thr Glu Ala Met Gln Ser Leu Gln Ile Gln

1 5 10 15

<210> 203

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 203

Val Ile Asp Ala Ser Asn Val Glu Lys Ala Phe Ile Ile Thr Asp Gly

1 5 10 15

<210> 204

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 204

Val Thr Asp Thr Ser Glu Ala Ala Ile Thr Asp Lys His Gly Asp Met

1 5 10 15

<210> 205

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 205

Tyr Ala Thr Glu Leu Ser Ser Leu Thr Thr Gln Val Ser Gln Leu Ile

1 5 10 15

<210> 206

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 206

Tyr Ala Thr Lys Ala Leu Asn Ser Val Lys Thr Ala Asn Asp Asp Gln

1 5 10 15

<210> 207

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 207

Tyr Ala Thr Lys Ser Leu Asn Ser Val Lys Thr Ser Asn Glu Glu Gln

1 5 10 15

<210> 208

<211> 16

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 208

Tyr Ala Thr Asn Ala Leu Asn Ser Val Arg Thr Ala Asn Glu Asp Gln

1 5 10 15

<210> 209

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 209

Ala Thr Glu Ala Leu Asn Glu Phe Lys Thr Gln Ile Thr Asp Leu Ala

1 5 10 15

Asp

<210> 210

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 210

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

1 5 10 15

Glu

<210> 211

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 211

Asp Asp Gly Ser Thr Ser Asp Gly Thr Ser Tyr Ala Thr Asn Val Leu

1 5 10 15

Lys

<210> 212

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 212

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

1 5 10 15

Tyr

<210> 213

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 213

Asp Phe Asn Pro Ser Lys Val Ala Gln Asp Pro Lys Val Thr Ala Tyr

1 5 10 15

Ile

<210> 214

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 214

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

1 5 10 15

Gln

<210> 215

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 215

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

1 5 10 15

Asn

<210> 216

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 216

Asp Phe Ser Gln Ser Glu Ile Ala Ala Ala Gly Ala Val Thr Asp Tyr

1 5 10 15

Ile

<210> 217

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 217

Glu Glu Glu Asp Pro Ala Asp Ala Ala Phe Pro Thr Leu Gln Ala Ser

1 5 10 15

Ala

<210> 218

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 218

Phe Asn Pro Ser Val Glu Ile Gly Ser Gln Gln Asn Val Thr Asp Tyr

1 5 10 15

Val

<210> 219

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 219

Gly Val Ile Asp Ser Ala Pro Val Val Gln Ser Ala Ile Thr Asn Gly

1 5 10 15

Gln

<210> 220

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 220

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

1 5 10 15

Thr

<210> 221

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 221

Met Asn Ser Leu Lys Thr Gln Ala Glu Glu Leu Val Met Thr Glu Leu

1 5 10 15

Ile

<210> 222

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 222

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

1 5 10 15

Lys

<210> 223

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 223

Thr Asp Ala Lys Asn Glu Leu Thr Thr Gln Val Thr Asp Leu Thr Thr

1 5 10 15

Gln

<210> 224

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 224

Thr Glu Gly Met Asn Ser Leu Lys Thr Leu Ala Ser Gln Leu Thr Asp

1 5 10 15

Gln

<210> 225

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 225

Thr Glu Ser Leu Gln Ser Leu Lys Ala Gln Ile Met Asp Leu Ile Asp

1 5 10 15

Gln

<210> 226

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 226

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

1 5 10 15

Asn

<210> 227

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 227

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

1 5 10 15

Gln

<210> 228

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 228

Val Phe Asp Ser Ser Ala Ala Asp Lys Ala Ile Gln Gly Asp Leu Lys

1 5 10 15

Ala

<210> 229

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 229

Val Thr Asp Thr Ser Ala Ile Glu Ala Ala Val Val Gln Ile Ile Thr

1 5 10 15

Asp

<210> 230

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 230

Tyr Ala Glu Ala Ser Leu Lys Thr Thr Ala Thr Arg Glu Asp Leu Ile

1 5 10 15

Glu

<210> 231

<211> 17

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 231

Met Asp Phe Asn Pro Ser Glu Val Ala Ser Gln Val Thr Asn Tyr Ile

1 5 10 15

Gln

<210> 232

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 232

Ala Ala Gly Asp Asp Pro Ala Val Ala Ala Val Gln Thr Ala Ala Thr

1 5 10 15

Glu Tyr

<210> 233

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 233

Ala Ala Thr Leu Arg Ile Phe Ala Ile Asp Thr Thr Ala Val Glu Ser

1 5 10 15

Ala Ile

<210> 234

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 234

Ala Ser Asp Gly Thr Ser Ser Thr Ser Tyr Glu Thr Val Pro Val Asn

1 5 10 15

Ser Leu

<210> 235

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 235

Ala Thr Gln Ala Ala Asn Asn Leu Ala Thr Gln Ala Thr Asn Leu Val

1 5 10 15

Asn Gln

<210> 236

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 236

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

1 5 10 15

Thr Asn

<210> 237

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 237

Asp Gly Asp Thr Ala Thr Ala Asn Ser Tyr Leu Thr Glu Ala Ala Asn

1 5 10 15

Ser Leu

<210> 238

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 238

Glu Asp Asp Asp Ala Val Lys Ala Ala Phe Asp Lys Leu Gln Ala Ser

1 5 10 15

Gln Thr

<210> 239

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 239

Glu Glu Glu Glu Ala Val Lys Ala Ala Phe Glu Lys Leu Gln Ala Ser

1 5 10 15

Gln Thr

<210> 240

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 240

Glu Asp Asp Glu Ala Val Lys Ser Ala Phe Asp Lys Leu Gln Ala Ser

1 5 10 15

Gln Thr

<210> 241

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 241

Glu Gly Asp Asp Asp Ala Ala Val Lys Ala Ala Phe Glu Lys Leu Gln

1 5 10 15

Ala Ser

<210> 242

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 242

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

1 5 10 15

Ala Ser

<210> 243

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 243

Phe Ser Pro Thr Glu Val Ala Ser Leu Leu Asp Val Gly Thr Asn Tyr

1 5 10 15

Val Gln

<210> 244

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 244

Ile Asp Thr Ser Ala Ile Glu Asn Ala Val Lys Lys Ser Asp Gly Gln

1 5 10 15

Gly Glu

<210> 245

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 245

Ile Asp Thr Ser Ala Ile Glu Thr Ala Val Arg Lys Ser Asp Gly Gln

1 5 10 15

Gly Glu

<210> 246

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 246

Met Asp Phe Leu Leu Asn Pro Glu Asn Ile Ala Ala Val Thr Asn Tyr

1 5 10 15

Ala Gln

<210> 247

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 247

Met Glu Phe Leu Tyr Lys Pro Asp Val Ala Ala Glu Leu Thr Asp Tyr

1 5 10 15

Ile Gln

<210> 248

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 248

Met Asp Tyr Leu Val Pro Ser Pro Thr Glu Val Pro Ser Met Val Thr

1 5 10 15

Asp Tyr

<210> 249

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 249

Met Phe Asn Pro Ser Val Glu Ile Gly Ser Gln Gln Asn Val Thr Asp

1 5 10 15

Tyr Val

<210> 250

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 250

Gln Ala Met Asp Ser Leu Lys Thr Ala Thr Ala Ser Val Ala Asp Leu

1 5 10 15

Leu Asp

<210> 251

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 251

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

1 5 10 15

Ile Gln

<210> 252

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 252

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

1 5 10 15

Leu Ile

<210> 253

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 253

Thr Asn Met Ser Thr Ala Tyr Ala Phe Leu Glu Ala Ile Asn Ser Leu

1 5 10 15

Lys Thr

<210> 254

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 254

Thr Ser Ala Val Ala Ser Ala Asp Thr Asp Gly Ser Asp Asp Gln Gly

1 5 10 15

Asp Met

<210> 255

<211> 18

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 255

Met Gln Ser Val Ile Thr Asp Val Thr Gly Gln Leu Thr Ala Val Gln

1 5 10 15

Ala Asp

<210> 256

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 256

Ala Glu Leu Gly Asp Asp Pro Ala Ala Ala Ile Asp Ala Leu Ala Ala

1 5 10 15

Ala Ala Thr

<210> 257

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 257

Ala Gly Val Ile Asn Thr Ala Val Ile Glu Gln Ala Ile Thr Asp Ala

1 5 10 15

Ser Asp Met

<210> 258

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 258

Asp Phe Tyr Pro Ser Glu Leu Ala Gly Gln Ile Asp Gln Leu Ala Asp

1 5 10 15

Tyr Ile Gln

<210> 259

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 259

Glu Ala Leu Ser Ser Leu Lys Ile Lys Ile Asp Asp Gln Pro Thr Asp

1 5 10 15

Leu Ile Asp

<210> 260

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 260

Glu Glu Met Ala Ala Ala Ala Asn Ser Leu Lys Thr Gln Ala Lys Glu

1 5 10 15

Leu Val Asp

<210> 261

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 261

Glu Gly Asp Asp Pro Leu Ser Arg Leu Ala Arg Ala Ser Leu Gln Ala

1 5 10 15

Ser Ala Thr

<210> 262

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 262

Phe Asn Pro Ser Glu Asn Ala Ser Lys Leu Thr Asn Arg Asn Ile His

1 5 10 15

Tyr Ile Gln

<210> 263

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 263

Ile Asp Thr Ser Ser Ala Ser Ser Ile Ala Ala Ala Glu Ala Ser Ile

1 5 10 15

Ala Ala Gly

<210> 264

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 264

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

1 5 10 15

Ser Val Ile

<210> 265

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 265

Met Glu Ile Asn Pro Ser Glu Val Thr Lys Ile Leu Lys Glu Gln Ile

1 5 10 15

Lys Asn Tyr

<210> 266

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 266

Met Glu Ile Asn Pro Ser Glu Val Thr Lys Ile Leu Lys Asp Gln Ile

1 5 10 15

Arg Asn Tyr

<210> 267

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 267

Met Gln Ser Leu Lys Thr Gln Glu Glu Glu Lys Arg Lys Ala Asp Leu

1 5 10 15

Val Asp Gln

<210> 268

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 268

Thr Ala Ile Thr Ser Tyr Ala Thr Glu Asn Ala Thr Gln Met Asn Ser

1 5 10 15

Ile Lys Thr

<210> 269

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 269

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

1 5 10 15

Gly Thr Asp

<210> 270

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 270

Thr Ser Leu Pro Ser Ile Ser Gly Gly Ile Glu Thr Ser Glu Ser Ala

1 5 10 15

Gly Thr Asp

<210> 271

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 271

Thr Thr Thr Gly Tyr Gly Met Asn Ser Leu Lys Ser Lys Glu Thr Asn

1 5 10 15

Leu Ile Asp

<210> 272

<211> 19

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 272

Ser Gly Val Gly Asp Gly Val Asp Val Val Ser Ala Ile Glu Gly Ala

1 5 10 15

Ala Gly Pro

<210> 273

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 273

Ala Asp Asp Gly Glu Gly Phe Thr Gly Ser Thr Thr Glu Ala Met Asn

1 5 10 15

Lys Leu Lys Thr

20

<210> 274

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 274

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

1 5 10 15

Ala Thr Glu Tyr

20

<210> 275

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 275

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

1 5 10 15

Thr Glu Gly Met

20

<210> 276

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 276

Ala Gly Glu Gly Thr Thr Thr Ala Thr Val Leu Thr Gln Ser Ile Ala

1 5 10 15

Thr Asp Gly Met

20

<210> 277

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 277

Ala Ser Val Ile Ser Thr Arg Asn Val Glu Thr Ala Met Lys Asn Gly

1 5 10 15

Gln Gly Asp Leu

20

<210> 278

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 278

Asp Asp Ala Ser Ser Thr Val Glu Ser Ala Thr Ser Ser Ala Thr Glu

1 5 10 15

Ala Met Glu Ser

20

<210> 279

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 279

Asp Asp Ala Thr Ser Gln Ala Lys Ala Ala Phe Asp Ser Leu Thr Ala

1 5 10 15

Gln Ala Thr Glu

20

<210> 280

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 280

Asp Asp Pro Asn Ala Thr Ala Ala Phe Glu Ser Leu Gln Ala Asp Ile

1 5 10 15

Ala Ala Asn Glu

20

<210> 281

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 281

Asp Thr Ser Ser Glu Ser Ala Ile Glu Ser Ala Val Gly Gln Gly Asp

1 5 10 15

Leu Ala Met Lys

20

<210> 282

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 282

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

1 5 10 15

Ala Ser Gln Thr

20

<210> 283

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 283

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

1 5 10 15

Thr Asp Phe Val

20

<210> 284

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 284

Met Gln Phe Asn Pro Tyr Glu Ile Ser Ser Gln Ile Lys Gln Asn Gly

1 5 10 15

Gly Tyr Ile Gln

20

<210> 285

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 285

Thr Ser Leu Pro Gly Val Ile Gly Ala Ala Ala Val Gln Ser Ala Lys

1 5 10 15

Thr Glu Gly Gln

20

<210> 286

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 286

Val Ile Asp Val Ser Ser Ala Ser His Val Gln Ser Phe Val Glu Ser

1 5 10 15

Ala Ile Thr Asp

20

<210> 287

<211> 20

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 287

Tyr Ser Thr Glu Asp Ala Ala Ala Met Asn Ala Ala Asp Gly Leu Lys

1 5 10 15

Thr Gln Ala Thr

20

<210> 288

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 288

Ala Ala Gly Asn Asp Arg Val Phe Thr Arg Arg Ala Phe Thr Met His

1 5 10 15

Pro Tyr Gly Val Lys

20

<210> 289

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 289

Ala Asp Gln Gly Thr Met Thr Ala Thr Asp Tyr Ala Thr Tyr Thr Ser

1 5 10 15

Leu Phe Glu Ala Met

20

<210> 290

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 290

Ala Pro Gly Glu Asp Pro Glu Ala Arg Ala Ala Tyr Asp Glu Met Gln

1 5 10 15

Ala Ala Ala Ala Glu

20

<210> 291

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 291

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

1 5 10 15

Leu Thr Asp Leu Val

20

<210> 292

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 292

Asp Gly Tyr Gly Lys Ala Thr Ser Phe Ser Thr Glu Ala Val Asn Ser

1 5 10 15

Leu Thr Asn Ile Ile

20

<210> 293

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 293

Gly Val Ile Asp Thr Ser Ala Val Glu Ser Ala Ile Thr Asp Gly Gln

1 5 10 15

Gly Asp Met Lys Ala

20

<210> 294

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 294

Leu Asp Phe Asn Ser Ser Glu Val Ala Leu Ser Glu Gly Met Gln Val

1 5 10 15

Thr Ser Phe Leu Gln

20

<210> 295

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 295

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

1 5 10 15

Ala Val Glu Ser Ala

20

<210> 296

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 296

Thr Gly Leu Pro His Ala Phe Pro Ala Val Ser Gly Ala Ser Ala Val

1 5 10 15

Glu Ser Ala Ile Thr

20

<210> 297

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 297

Thr Gly Leu Pro His Ala Phe Pro Ala Val Ser Gly Ala Ser Ala Val

1 5 10 15

Glu Ser Ala Ile Thr

20

<210> 298

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 298

Thr Ser Ala Glu Ala Thr Tyr Glu Lys Ala Met Asn Ala Leu Lys Ile

1 5 10 15

Gln Ala Ile Asp Leu

20

<210> 299

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 299

Ala Glu Gly Asp Asp Pro Ala Lys Ala Ala Phe Asp Ser Leu Gln Ala

1 5 10 15

Ser Ala Thr Glu Tyr

20

<210> 300

<211> 21

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 300

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

1 5 10 15

Ser Ala Thr Glu Tyr

20

<210> 301

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 301

Ala Glu Gly Asp Ala Ser Ser Gln Ala Lys Ala Ala Phe Asp Ser Leu

1 5 10 15

Thr Ala Gln Ala Thr Glu

20

<210> 302

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 302

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

1 5 10 15

Thr Ala Gln Ala Thr Glu

20

<210> 303

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 303

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

1 5 10 15

Thr Ala Gln Ala Thr Glu

20

<210> 304

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 304

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

1 5 10 15

Thr Glu Gly Met Lys Ser

20

<210> 305

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 305

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

1 5 10 15

Ile Asp Leu Ile Ser Gln

20

<210> 306

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 306

Ala Thr Pro Leu Pro Ala Phe Thr Val Asp Val Glu Ser Gly Ile Thr

1 5 10 15

Asp Pro Thr Ser Gly Gln

20

<210> 307

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 307

Asp Gly Ser Thr Ser Tyr Thr Leu Asp Thr Leu Asp Ser Leu Lys Thr

1 5 10 15

Gln Gly Thr Glu Ile Ile

20

<210> 308

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 308

Glu Gly Ser Asp Pro Ala Ala Thr Val Phe Asp Ser Leu Gln Ala Ala

1 5 10 15

Lys Ala Arg Gly Thr Glu

20

<210> 309

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 309

Gly Thr Ser Thr Thr Ala Phe Tyr Asp Arg Ala Thr Ile Asp Met Asn

1 5 10 15

Ala Leu Arg Thr Gln Ala

20

<210> 310

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 310

Gly Val Asp Glu Leu Trp Val Arg Arg Gln Phe Val Met His Pro Tyr

1 5 10 15

Gly Ile Lys Trp Thr Asp

20

<210> 311

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 311

Ile Ile Asp Ala Gly Ala Val Ile Ser Gly Ile Thr Asp Gly Tyr Gln

1 5 10 15

Gly Ser Ala Pro Asp Met

20

<210> 312

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 312

Ile Ile Glu Ala Gly Thr Val Ile Ser Gly Ile Thr Glu Gly Tyr Gln

1 5 10 15

Gly Ser Ser Pro Asp Met

20

<210> 313

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 313

Thr Ser Thr Ala Thr Gly Pro Ile Ile Tyr Ala Ala Glu Ala Ile Asn

1 5 10 15

Ser Gly Ala Asp Leu Ile

20

<210> 314

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 314

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

1 5 10 15

Lys Thr Ser Thr Ala Leu

20

<210> 315

<211> 22

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 315

Ala Asp Asp Ala Thr Ser Gln Ala Lys Ala Ala Phe Asp Ser Leu Thr

1 5 10 15

Ala Gln Ala Thr Glu Met

20

<210> 316

<211> 23

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 316

Ala Asp Asp Gly Asp Asp Thr Thr Thr Ala Thr Thr Ser Ala Thr Glu

1 5 10 15

Pro Val Ala Ser Ile Lys Thr

20

<210> 317

<211> 23

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 317

Ala Asp Ser Ser Thr Asp Tyr Ala Gly Gln Ala Met Asp Ser Leu Leu

1 5 10 15

Thr Gln Ala Asn Asp Leu Ile

20

<210> 318

<211> 23

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 318

Ala Phe Thr Gln Pro Ile Asp Thr Ser Ala Ile Glu Thr Ala Ile Ile

1 5 10 15

Asp Gln Ser Lys Gly Gln Gly

20

<210> 319

<211> 23

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 319

Ala Gly Asp Gly Thr Thr Thr Ala Thr Val Leu Ala Tyr Ala Met Val

1 5 10 15

Arg Glu Gly Met Lys Asn Leu

20

<210> 320

<211> 23

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 320

Ser Thr Gly Gly Thr Asp Tyr Ala Gly Gln Ala Met Asp Ala Leu Leu

1 5 10 15

Thr Gln Ala Asn Asp Leu Ile

20

<210> 321

<211> 23

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 321

Thr Ala Thr Asn Tyr Ala Thr Glu Ala Met Thr Ser Leu Lys Thr Gln

1 5 10 15

Ala Thr Asp Leu Ile Ala Gln

20

<210> 322

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 322

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

1 5 10 15

Lys Glu Ser Met Asn Ser Ile Lys

20

<210> 323

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 323

Ala Lys Gly Asn Asp Asn Val Phe Thr Arg Arg Ala Phe Val Met His

1 5 10 15

Pro Tyr Gly Val Lys Trp Thr Asp

20

<210> 324

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 324

Ala Asn Asp Gly Thr Ala Thr Ala Thr Ser Ile Glu Thr Gly Ala Thr

1 5 10 15

Glu Ala Leu Pro Thr Ser Ala Thr

20

<210> 325

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 325

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

1 5 10 15

Val Leu Ala Ser Gln Ala Thr Asp

20

<210> 326

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 326

Ala Thr Glu Ala Met Asn Leu Trp Ile Arg Glu Lys Thr Gln Glu Ala

1 5 10 15

Ile Ala Arg Asp Ala Leu Asp Gln

20

<210> 327

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 327

Asp Asp Gly Thr Ala Thr Ser Tyr Ser Val Ala Leu Ser Leu Asn Pro

1 5 10 15

Tyr Ser Thr Glu Ala Met Asp Ser

20

<210> 328

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 328

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

1 5 10 15

Ala Glu Ala Thr Thr Leu Glu Asp

20

<210> 329

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 329

Asp Asn Pro Ser Lys Ala Ala Tyr Asp Ser Leu Gln Ala Gln Leu Lys

1 5 10 15

Arg Asp Ile Glu Ala Ala Glu Tyr

20

<210> 330

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 330

Asp Asn Pro Ser Lys Ala Ala Tyr Asp Ser Leu Gln Ala Gln Leu Lys

1 5 10 15

Arg Asp Leu Glu Ala Ser Glu Tyr

20

<210> 331

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 331

Glu Ala Met Ser Thr Tyr Arg Ala Met Arg Tyr Leu Lys Thr Asp Glu

1 5 10 15

Val Ser Thr Asp Leu Ile Glu Gln

20

<210> 332

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 332

Met Thr Phe Asn Pro Ala Ala Val Val Ser Pro Leu Leu Leu Gly Asn

1 5 10 15

Ala Ser Arg Ile Thr Asn Tyr Ile

20

<210> 333

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 333

Ser Leu Pro Ala Phe Ile Glu Val Ile Gly Thr Ser Gly Val Asp Ala

1 5 10 15

Ala Ile Lys Gln Tyr Ala Thr Asp

20

<210> 334

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 334

Thr Asp Leu Pro Thr Ile Asp Glu Leu Gly Ser Ala Gly Leu Ile Asp

1 5 10 15

Thr Ser Thr Val Asp Ser Gln Ile

20

<210> 335

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 335

Thr Pro Leu Gln Ala Phe Ser Gly Val Pro Arg Thr Arg Asp Thr Ser

1 5 10 15

Glu Ser Pro Ile Thr Asp Gly Glu

20

<210> 336

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 336

Thr Ser Phe Ala Glu Asp Ala Met Ser Ser Leu Thr Thr Ala Gly Pro

1 5 10 15

Thr Thr Ala Thr Asp Leu Leu Asp

20

<210> 337

<211> 24

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 337

Thr Ser Met Pro Ala Tyr Thr Ser Lys Ala Leu Val Ile Asp Thr Ser

1 5 10 15

Pro Ile Glu Lys Glu Ile Ser Asn

20

<210> 338

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 338

Ala Phe Ser Ala Val Leu Asp Thr Ala Val Thr Ala Val Glu Asn Val

1 5 10 15

Asn Ala Ala Ile Glu Ala Ile Thr Asp

20 25

<210> 339

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 339

Ala Gly Glu Gly Ala Ser Thr Asp Tyr Ala Gly Gln Ala Met Asp Ser

1 5 10 15

Leu Leu Thr Gln Ala Asn Asp Leu Ile

20 25

<210> 340

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 340

Ala Thr Pro Val Phe Val Asp Val Leu Gly Asn Gln Leu Ile Asp Pro

1 5 10 15

Asn Ala Ile Glu Lys Ala Ile Thr Asp

20 25

<210> 341

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 341

Ala Thr Val Pro Ala Phe Ala Ala Val Val Asp Thr Ala Ala Ile Gln

1 5 10 15

Ala Gln Ile Gln Glu Gly Gln Gly Asp

20 25

<210> 342

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 342

Ala Thr Val Pro Ala Phe Ala Ala Val Val Asp Thr Ala Ala Ile Gln

1 5 10 15

Thr Gln Ile Gln Glu Gly Gln Ala Asp

20 25

<210> 343

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 343

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

1 5 10 15

Leu Leu Glu Ala Met Pro Gly Leu Lys

20 25

<210> 344

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 344

Glu Asp Val Thr Val Thr Ala Thr Ile Ser Tyr Ala Ala Ser Pro Met

1 5 10 15

Asn Gly Gln Val Val Asn Leu Ile Asn

20 25

<210> 345

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 345

Gly Ile Thr Asp Thr Ser Asp Val Ser Leu Val Ala Leu Ser Asp Ala

1 5 10 15

Val Glu Ser Ala Ile Thr Glu Thr Gln

20 25

<210> 346

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 346

Thr Ser Asn Ser Ala Glu Ser Met Asn Ser Leu Ser Lys His Pro Gln

1 5 10 15

Lys Thr Pro Ile Thr Gln Leu Ile Asp

20 25

<210> 347

<211> 25

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 347

Ala Glu Pro Asn Ala Ala Thr Asn Tyr Ala Thr Glu Ala Met Asp Ser

1 5 10 15

Leu Lys Thr Gln Ala Ile Asp Leu Ile

20 25

<210> 348

<211> 26

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 348

Ala Thr Ser Ser Pro Ser Tyr Ala Val Ile Asp Val Thr Ala Val Thr

1 5 10 15

Thr Gly Ile Thr Asp Ala Gln Thr Ala Met

20 25

<210> 349

<211> 26

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 349

Asp Asp Gly Thr Val Ser Arg Phe Ala Glu Asp His Phe Leu Ile Thr

1 5 10 15

Thr Thr Thr Ala Tyr Ala Thr Glu Ala Met

20 25

<210> 350

<211> 26

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 350

Ser Leu Phe Gly Pro Val Phe Ala Gly Ile Ile Thr Ala Ala Val Leu

1 5 10 15

Ser Ala Ile Met Ser Thr Ala Asp Ser Gln

20 25

<210> 351

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 351

Ala Ala Asp Gly Ala Thr Gly Ser Thr Asp Tyr Ala Gly Gln Ala Met

1 5 10 15

Asp Ser Leu Leu Thr Gln Ala Asn Asp Leu Ile

20 25

<210> 352

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 352

Ala Ala Asp Gly Ala Thr Thr Asp Tyr Ala Thr Asp Ala Phe Ala Thr

1 5 10 15

Leu Lys Thr Thr Ala Thr Asp Met Ile Glu Gln

20 25

<210> 353

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 353

Ala Ala Asp Gly Ser Thr Gly Gly Ala Asp Tyr Ala Gly Gln Ala Met

1 5 10 15

Asp Ala Leu Leu Thr Gln Ala Asn Asp Leu Ile

20 25

<210> 354

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 354

Ala Ala Glu Gly Ala Thr Gly Gly Thr Asp Tyr Ala Gly Gln Ala Met

1 5 10 15

Asp Ala Leu Leu Thr Gln Ala Asn Asp Leu Ile

20 25

<210> 355

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 355

Ala Ala Glu Gly Ala Thr Gly Ser Thr Asp Tyr Ala Gly Lys Ala Met

1 5 10 15

Asp Ser Leu Leu Thr Gln Ala Asn Asp Leu Ile

20 25

<210> 356

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 356

Ala Ala Glu Gly Ala Thr Gly Ser Thr Asp Tyr Ala Gly Gln Ala Met

1 5 10 15

Asp Ser Leu Leu Thr Gln Ala Asn Asp Leu Ile

20 25

<210> 357

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 357

Ala Glu Gly Gly Ala Ala Thr Thr Pro Asp Tyr Ala Gly Gln Ala Met

1 5 10 15

Asp Ser Leu Leu Thr Gln Ala Asn Thr Leu Ile

20 25

<210> 358

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 358

Ala Glu Pro Asp Glu Glu Ala Lys Ala Met Phe Asp Ser Leu Ala Lys

1 5 10 15

Pro Ala Leu Gly Pro Gly Ala Ser Ala Thr Glu

20 25

<210> 359

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 359

Ala Glu Pro Asp Glu Glu Ala Lys Ala Met Phe Glu Ser Leu Ala Lys

1 5 10 15

Pro Ala Leu Gly Pro Gly Ala Ser Ala Thr Asp

20 25

<210> 360

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 360

Ala Phe Thr Gly Thr Ile Asp Val Asp Ser Ala Ala Glu Ser Leu Lys

1 5 10 15

Leu Phe Thr Lys Thr Phe Ala Ile Thr Asp Gly

20 25

<210> 361

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 361

Ala Thr Ala Leu Pro Ala Trp Ala Ala Val Ile Asp Thr Thr Glu Val

1 5 10 15

Gln Ala Thr Phe Gly Glu Ala Lys Gly Asp Met

20 25

<210> 362

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 362

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

1 5 10 15

Glu Ala Ala Ile Thr Asp Gly Lys Gly Asp Met

20 25

<210> 363

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 363

Ala Thr Ser Leu Pro Ala Phe Ala Gly Val Ile Gly Ala Val Ala Leu

1 5 10 15

Ala Leu Ala Val Gly Asp Gly Asp Met Lys Ala

20 25

<210> 364

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 364

Ala Thr Thr Leu Pro Ala Val Ala Thr Arg Ala Gly Asp Thr Trp Ala

1 5 10 15

Val Glu Val Arg Ile Ser Asp Gly Glu Asp Met

20 25

<210> 365

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<220>

<221> misc_feature

<222> (14)..(14)

<223> Xaa can be any naturally occurring amino acid

<400> 365

Thr Ser Val Pro Ala Phe Ala Ala Ser Val Ile Asp Thr Xaa Ala Val

1 5 10 15

Glu Gln Ala Ile Thr Asp Gly Lys Ser Asp Met

20 25

<210> 366

<211> 27

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 366

Thr Ser Val Pro Ala Phe Ala Gly Ser Val Ile Asp Thr Ser Ala Val

1 5 10 15

Glu Thr Ala Ile Thr Asp Gly Lys Ser Asp Met

20 25

<210> 367

<211> 28

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 367

Ala Asp Asp Gly Thr Ser Thr His Ile Ser Tyr Met Gln Leu His Gly

1 5 10 15

Arg Val Cys Arg Phe Ala Asn Ala Leu Lys Ala Gln

20 25

<210> 368

<211> 28

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 368

Ala Phe Ala Asn Asp Asp Ala Ile Asp Thr Ala Val Gly Thr Ala Gln

1 5 10 15

Asn Val Thr Asp Thr Trp Leu Ala Gln Gly Asp Met

20 25

<210> 369

<211> 28

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 369

Ala Ser Ser Met Pro Val Trp Ala Ala Ser Val Ile Asp Thr Ser Ser

1 5 10 15

Val Glu Gln Ala Ile Thr Asp Gly Lys Gly Asp Met

20 25

<210> 370

<211> 28

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 370

Ala Ser Ser Val Pro Ala Trp Ala Gly Ser Val Ile Asp Thr Ser Ala

1 5 10 15

Val Glu Ser Ala Ile Ser Asp Gly Lys Gln Asp Met

20 25

<210> 371

<211> 28

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 371

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

1 5 10 15

Glu Thr Ala Met Thr Asp Gly Gln Gly Asp Met Lys

20 25

<210> 372

<211> 28

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 372

Thr Ala Leu Pro Val Ile Glu Thr Ala Ala Gly Asp Val Ser Ala Phe

1 5 10 15

Val Pro Thr Asn Val Ile Ser Ile Thr Asp Gly Gln

20 25

<210> 373

<211> 28

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 373

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

1 5 10 15

Ile Pro Thr Asn Val Ile Ser Ile Thr Asp Gly Gln

20 25

<210> 374

<211> 29

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 374

Ala Glu Asp Pro Ala Met Gln Gln Arg Ser Val Ala Ala Phe Asp Ala

1 5 10 15

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

20 25

<210> 375

<211> 29

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 375

Ala Glu Asp Pro Ala Met Gln Gln Arg Ser Val Ala Ala Phe Asp Ala

1 5 10 15

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

20 25

<210> 376

<211> 29

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 376

Ala Glu Asp Pro Ala Met Gln Gln Arg Ser Val Ala Ala Phe Glu Ala

1 5 10 15

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

20 25

<210> 377

<211> 29

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 377

Ala Ser Thr Leu Pro Ala Ile Ala Thr Arg Ala Gly Asp Val Trp Thr

1 5 10 15

Val Glu Val Arg Ile Thr Asp Gly Glu Asp Met Ser Ala

20 25

<210> 378

<211> 29

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 378

Asp Phe Asn Ile Ala Pro Gly Leu Ala Pro Leu Asn Gly Leu Leu Leu

1 5 10 15

Pro Ser Glu Val Val Ser Gln Ile Ser Ser Phe Ile Gln

20 25

<210> 379

<211> 29

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 379

Ser Thr Thr Thr Ser Tyr Ser Ser Glu Leu Ser Val Tyr Pro Ser Leu

1 5 10 15

Asn Ser Leu Lys Glu Thr His Gly Gln Asp Leu Ile Asp

20 25

<210> 380

<211> 29

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 380

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

1 5 10 15

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

20 25

<210> 381

<211> 29

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 381

Ala Asp Asp Gly Thr Ser Thr Ala Thr Ser Tyr Ala Thr Glu Ala Met

1 5 10 15

Asn Ser Leu Lys Thr Gln Ala Thr Asp Leu Ile Asp Gln

20 25

<210> 382

<211> 30

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 382

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

1 5 10 15

Thr Ser Asn Ser Gly Pro Trp Thr Thr His Ser Thr Asn Leu

20 25 30

<210> 383

<211> 31

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 383

Met Phe Asn Pro Ser Glu Val Ala Asp Asn Leu Lys Arg Ser Gly Ala

1 5 10 15

Phe Val Pro Gly Ile Arg Pro Gly Gln Gln Thr Ala Asn Tyr Ile

20 25 30

<210> 384

<211> 32

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 384

Asp His Leu Glu Thr Arg Leu Ala Phe Asn Tyr His Pro Phe Gly His

1 5 10 15

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

20 25 30

<210> 385

<211> 33

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 385

Ala Gly Asp Gly Thr Thr Thr Ala Thr Ile Tyr Ala Glu Ala Ile Phe

1 5 10 15

Asn Glu Gly Leu Lys Asn Val Val Ala Gly Ala Asp Ala Met Ser Leu

20 25 30

Lys

<210> 386

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 386

Gly Asp Asp Ile Leu Ile Asn Arg Gln Ala Phe Val Leu His Pro Arg

1 5 10 15

Gly Val Glu Phe Lys Asn Ala Ser Val Ala Gly Ala Thr Pro Ser Asn

20 25 30

Ala Glu Val

35

<210> 387

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 387

Gly Asp Asp Ile Leu Ile Asn Arg Gln His Phe Ile Leu His Pro Arg

1 5 10 15

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

20 25 30

Ala Glu Leu

35

<210> 388

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 388

Gly Asp Asp Ile Leu Ile Asn Arg Gln His Phe Leu Leu His Pro Arg

1 5 10 15

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

20 25 30

Ala Glu Leu

35

<210> 389

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 389

Gly Asp Asp Ile Leu Ile Asn Arg Gln His Phe Leu Leu His Pro Arg

1 5 10 15

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

20 25 30

Ala Glu Leu

35

<210> 390

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 390

Gly Asp Asp Ile Leu Ile Asn Arg Gln His Phe Leu Leu His Pro Arg

1 5 10 15

Gly Val Lys Phe Thr Asp Lys Ser Val Ser Gly Ser Ser Pro Thr Asn

20 25 30

Ala Glu Leu

35

<210> 391

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 391

Gly Asp Asp Ile Leu Ile Asn Arg Gln His Phe Leu Leu His Pro Arg

1 5 10 15

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

20 25 30

Ala Glu Leu

35

<210> 392

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 392

Gly Asp Asp Ile Leu Val Asn Arg Gln His Phe Leu Leu His Pro Arg

1 5 10 15

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

20 25 30

Ala Glu Leu

35

<210> 393

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 393

Gly Asp Asp Ile Leu Val Asn Arg Arg His Phe Val Leu His Pro Arg

1 5 10 15

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

20 25 30

Ala Glu Leu

35

<210> 394

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 394

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

1 5 10 15

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

20 25 30

Ala Glu Leu

35

<210> 395

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 395

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

1 5 10 15

Gly Ile Lys Phe Thr Asn Ala Thr Val Ala Lys Thr Ala Pro Thr Asn

20 25 30

Ala Glu Leu

35

<210> 396

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 396

Gly Glu Asp Ile Leu Ile Asn Arg Gln His Phe Val Leu His Pro Arg

1 5 10 15

Gly Val Ala Phe Gln Asn Ala Ser Val Ala Gly Ser Ser Pro Thr Asn

20 25 30

Ala Glu Leu

35

<210> 397

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 397

Gly Glu Asp Tyr Leu Ile Asn Arg Arg Thr Phe Ile Leu His Pro Arg

1 5 10 15

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

20 25 30

Ala Glu Leu

35

<210> 398

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 398

Gly Gly Asp Ile Leu Tyr Thr Arg Arg Gln Phe Val Leu His Pro Tyr

1 5 10 15

Gly Ile Ala Trp Gln Asp Ala Ser Val Ala Ala Glu Phe Pro Thr Asn

20 25 30

Val Glu Leu

35

<210> 399

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 399

Gly Gly Asp Ile Leu Tyr Thr Arg Arg Gln Phe Val Leu His Pro Tyr

1 5 10 15

Gly Ile Lys Trp Thr Asp Ala Ser Val Ala Gly Glu Phe Pro Thr Thr

20 25 30

Ala Glu Met

35

<210> 400

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 400

Gly Asn Asp Lys Leu Tyr Thr Arg Arg Ala Ile Val Met His Pro Tyr

1 5 10 15

Gly Leu Ser Phe Glu Pro Glu Glu Leu Ser Asp Phe Thr Pro Thr Asn

20 25 30

Gly Asp Leu

35

<210> 401

<211> 35

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 401

Gly Val Asp Tyr Leu Ile Thr Arg Arg His Phe Leu Leu His Pro Arg

1 5 10 15

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

20 25 30

Ala Glu Leu

35

<210> 402

<211> 36

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 402

Gly Ile Asp Tyr Leu Ile Asn Arg Lys Thr Phe Ile Leu His Pro Arg

1 5 10 15

Gly Val Lys Phe Thr Asn Thr Val Arg Ala Asn Thr Glu Thr Val Ser

20 25 30

Arg Ala Glu Leu

35

<210> 403

<211> 36

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 403

Gly Val Asp Tyr Leu Ile Asn Arg Lys Ala Phe Ile Leu His Pro Arg

1 5 10 15

Gly Ile Ala Tyr Thr Gly Ala Lys Arg Gly His Val Glu Thr Pro Thr

20 25 30

Arg Ala Glu Leu

35

<210> 404

<211> 36

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 404

Gly Val Asp Tyr Leu Ile Asn Arg Lys Thr Phe Ile Leu His Pro Arg

1 5 10 15

Gly Ile Lys Phe Thr Gly Ala Val Arg Ala Asn Gln Glu Thr Val Ser

20 25 30

Arg Ala Glu Leu

35

<210> 405

<211> 36

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 405

Gln Gly Asp Asp Ile Leu Ile Asn Arg Gln His Phe Ile Leu His Pro

1 5 10 15

Arg Gly Val Glu Phe Lys Asn Ala Ala Val Ala Gly Pro Ser Pro Thr

20 25 30

Asn Ala Glu Leu

35

<210> 406

<211> 36

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 406

Gln Gly Asp Asp Ile Leu Ile Asn Arg Gln His Phe Ile Leu His Pro

1 5 10 15

Arg Gly Val Glu Phe Lys Asn Ala Ala Val Ala Ser Ser Ser Pro Thr

20 25 30

Asn Thr Glu Leu

35

<210> 407

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 407

Ala Ala Gly Ile Asp Glu Ile Phe Thr Arg Arg Ala Phe Val Tyr His

1 5 10 15

Pro Tyr Gly Val Lys Phe Thr Ser Lys Ala Val Ala Gly Leu Thr Pro

20 25 30

Ser Asn Ala Glu Leu

35

<210> 408

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 408

Ala Ala Gly Asn Asp Arg Ile Phe Thr Arg Arg Ala Leu Val Met His

1 5 10 15

Pro Tyr Gly Ile Lys Trp Thr Asp Thr Ser Val Glu Gly Ala Thr Pro

20 25 30

Ser Asn Glu Glu Leu

35

<210> 409

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 409

Ala Ala Gly Asn Asp Arg Val Phe Thr Arg Arg Ala Phe Thr Met His

1 5 10 15

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

20 25 30

Ser Asn Ala Glu Met

35

<210> 410

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 410

Ala Ala Gly Val Asp Glu Leu Phe Thr Arg Arg Ala Phe Val Tyr His

1 5 10 15

Pro Tyr Gly Val Lys Phe Thr Ser Lys Ala Val Ala Gly Leu Thr Pro

20 25 30

Ser Asn Thr Glu Leu

35

<210> 411

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 411

Ala Ala Gly Val Asp Glu Val Phe Thr Arg Arg Ala Phe Val Phe His

1 5 10 15

Pro Tyr Gly Val Lys Phe Thr Asp Thr Thr Val Ala Gly Leu Thr Pro

20 25 30

Ser Asn Thr Glu Leu

35

<210> 412

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 412

Ala Ala Gly Val Asp Glu Val Phe Thr Arg Arg Ala Phe Val Tyr His

1 5 10 15

Pro Tyr Gly Ile Lys Phe Lys Ser Thr Thr Val Thr Gly Glu Thr Pro

20 25 30

Ser Asn Ala Glu Leu

35

<210> 413

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 413

Ala Ala Gly Val Asp Glu Val Tyr Thr Arg Arg Ala Phe Val Phe His

1 5 10 15

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

20 25 30

Ser Asn Ala Glu Leu

35

<210> 414

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 414

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

1 5 10 15

Pro Tyr Gly Val Arg Trp Gln Asp Asn Ser Ile Val Gly Leu Thr Pro

20 25 30

Ser Asn Ala Glu Leu

35

<210> 415

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 415

Gly Gln Asp Tyr Leu Val Thr Arg Arg His Tyr Val Leu His Pro Arg

1 5 10 15

Gly Ile Lys Trp Asp Pro Gly Ser Gly Val Pro Ala Ser Val Thr Pro

20 25 30

Ser Asp Ala Glu Leu

35

<210> 416

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 416

Ser Lys Gly Ile Asp Glu Val Tyr Thr Arg Arg Ala Phe Val Phe His

1 5 10 15

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

20 25 30

Ser Asn Ala Glu Leu

35

<210> 417

<211> 37

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 417

Ser Ser Gly Glu Asp Phe Leu Ile Asn Arg Gln Ile Tyr Ile Leu His

1 5 10 15

Pro Arg Gly Val Lys Phe Thr Asp Thr Ala Val Ala Asp Val Phe Pro

20 25 30

Thr Asn Ala Glu Leu

35

<210> 418

<211> 38

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 418

Lys Gly Asn Asp Glu Val Tyr Thr Arg Arg Ala Phe Thr Met His Pro

1 5 10 15

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

20 25 30

Pro Thr Asn Lys Asp Leu

35

<210> 419

<211> 38

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 419

Lys Gly Asn Asp Glu Val Tyr Thr Arg Arg Ala Phe Thr Met His Pro

1 5 10 15

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

20 25 30

Pro Thr Asn Asn Asp Leu

35

<210> 420

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 420

Ala Lys Gly Asn Asp Met Ile Tyr Thr Arg Arg Ala Leu Val Met His

1 5 10 15

Pro Tyr Gly Val Lys Trp Thr Gly Ala Glu Val Asp Ala Gly Asn Ile

20 25 30

Thr Pro Ser Asn Ala Asp Leu

35

<210> 421

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 421

Ala Lys Gly Asn Asp Gln Val Phe Thr Arg Arg Ala Phe Thr Met His

1 5 10 15

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

20 25 30

Thr Pro Thr Asn Ala Asp Leu

35

<210> 422

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 422

Ala Lys Gly Asn Asp Gln Val Phe Thr Arg Arg Ala Phe Thr Met His

1 5 10 15

Pro Tyr Gly Val Lys Phe Thr Asn Ala Asp Arg Glu Ala Gly Glu Ile

20 25 30

Thr Pro Thr Asn Lys Asp Leu

35

<210> 423

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 423

Ala Lys Gly Asn Asp Gln Val Phe Thr Arg Arg Ala Ile Thr Met His

1 5 10 15

Pro Tyr Gly Ile Lys Trp Thr Asp Asn Leu Arg Asp Asp Gly Asn Ile

20 25 30

Thr Pro Thr Asn Val Asp Leu

35

<210> 424

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 424

Ala Lys Gly Asn Asp Gln Val Phe Thr Arg Arg Ala Ile Thr Met His

1 5 10 15

Pro Tyr Gly Ile Lys Trp Thr Asp Asn Leu Arg Glu Glu Gly Asn Ile

20 25 30

Thr Pro Thr Asn Val Asp Leu

35

<210> 425

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 425

Ala Lys Gly Asn Asp Gln Val Phe Thr Arg Arg Ala Ile Thr Met His

1 5 10 15

Pro Tyr Gly Ile Lys Trp Thr Glu Asn Leu Arg Glu Glu Gly Asn Ile

20 25 30

Thr Pro Thr Asn Val Asp Leu

35

<210> 426

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 426

Ala Lys Gly Asn Asp Arg Ile Phe Thr Arg Trp Ala Leu Thr Met His

1 5 10 15

Pro Tyr Gly Val Lys Trp Asn Asp Ser Thr Arg Glu Asp Gly Asn Ile

20 25 30

Thr Pro Thr Asn Val Asp Leu

35

<210> 427

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 427

Ala Lys Gly Asn Asn Asn Ile Phe Thr Arg Arg Ala Phe Thr Met His

1 5 10 15

Pro Tyr Gly Val Lys Phe Thr Asn Ala Ala Arg Glu Ser Gly Phe Ile

20 25 30

Thr Pro Thr Asn Lys Asp Leu

35

<210> 428

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 428

Ala Lys Gly Thr Asp Ile Ile Tyr Thr Arg Arg Ala Val Thr Met His

1 5 10 15

Pro Tyr Gly Val Lys Trp Lys Asp Ala Glu Arg Glu Ala Gly Asn Met

20 25 30

Thr Pro Thr Asn Val Asp Leu

35

<210> 429

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 429

Asn Lys Gly Asn Asp Glu Val Tyr Thr Arg Arg Ala Phe Thr Met His

1 5 10 15

Pro Tyr Gly Val Lys Phe Thr Asn Thr Asp Arg Glu Ser Gly Glu Ile

20 25 30

Thr Pro Thr Asn Lys Asp Leu

35

<210> 430

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 430

Ser Lys Gly Asn Asp Gln Val Phe Thr Arg Arg Ala Phe Thr Met His

1 5 10 15

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

20 25 30

Thr Pro Thr Asn Ala Asp Leu

35

<210> 431

<211> 39

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 431

Ser Lys Gly Asn Asp Gln Val Phe Thr Arg Arg Ala Phe Thr Met His

1 5 10 15

Pro Tyr Gly Val Lys Phe Lys Asn Ala Val Arg Asp Ala Asn Asp Ile

20 25 30

Thr Pro Thr Asn Ala Glu Leu

35

<210> 432

<211> 40

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 432

Asp Gly Thr Ser Tyr Val Ser Ser Thr Ala Ser Pro Tyr Ser Ala Thr

1 5 10 15

Glu Asn Ser Thr Leu Lys Gly Ile Glu Leu Ile Tyr Lys Phe Pro Leu

20 25 30

Ala Ala Met Asn Ser Leu Lys Thr

35 40

<210> 433

<211> 44

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 433

Asp Asp Gly Ser Ser Thr Ala Ser Ala Ala Ser Val Ser Tyr Arg Val

1 5 10 15

Asp Gly Ala Pro Gly Ala Gln Val Ala Thr Ile Thr Leu Glu Arg Pro

20 25 30

Glu Ala Met Asn Gly Leu Asp Thr Glu Thr Lys Asp

35 40

<210> 434

<211> 50

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 434

Tyr Val Ile Thr Gly Ser Val Asn Pro Lys Arg Phe Ala Tyr Thr Glu

1 5 10 15

Arg Gly Asn Thr Gln Ile Val Ile Arg Glu Pro Phe Thr Asp His Pro

20 25 30

Ile Tyr Asp Ala Phe Lys Asp Cys Phe Tyr Asp Ala Tyr Asp Leu Glu

35 40 45

Leu Asp

50

<210> 435

<211> 67

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 435

Tyr Val Phe Gln Gly Gln Gly Asn Asp His Arg Phe Thr Thr Arg Pro

1 5 10 15

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

20 25 30

Asn Met Ile Arg Thr Leu Thr Ser Ala Val Pro Ile Ile Thr Gly Tyr

35 40 45

Glu Val Phe Thr Tyr Trp Ala Phe Ala Asn Gly Tyr Leu Gly Leu Phe

50 55 60

Asp Leu Gly

65

<210> 436

<211> 69

<212> PRT

<213> Artificial Sequence

<220>

<223> synthetic

<400> 436

Tyr Val Phe Gln Gly Gln Gly Asn Asp His Arg Phe Thr Thr Arg Pro

1 5 10 15

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

20 25 30

Asn Met Ile Arg Thr Leu Ala Ser Ala Val Pro Ile Ile Thr Gly Tyr

35 40 45

Glu Val Phe Thr Tyr Trp Ala Phe Ala Asn Gly Tyr Leu Gly Leu Phe

50 55 60

Asp Leu Gly Ser Ser

65