Interleukin-2 derivatives

Abstract

The present application discloses an IL-2 derivative, by introducing at least one cysteine residue on the basis of IL-2, partially or completely closing the binding plane of the IL-2 derivative and the alpha receptor subunit, while basically retaining the affinity to the beta and gamma receptor subunit complex. The present application also discloses a complex, including an IL-2 derivative, a third cysteine residue introduced on the basis of IL-2; a blocking module, the blocking module has or introduces a fourth cysteine residue; the third cysteine residue on the IL-2 derivative and the fourth cysteine residue on the blocking module form an intermolecular disulfide bond, thereby forming a complex of IL-2 and the blocking module, partially or completely closing the binding plane of the IL-2 derivative and the alpha receptor subunit, while basically retaining the affinity to the beta and gamma receptor subunit complex.

Description

Technical Field

[0001] This invention belongs to the field of molecular biology, specifically relating to an interleukin-2 derivative and its complex. Background Technology

[0002] Interleukin-2 (IL-2), discovered in 1976 and initially known as T cell growth factor (TCGF), is a globular glycoprotein that plays a crucial role in maintaining the normal function of T lymphocytes and NK cells. Natural IL-2 is a polypeptide consisting of 133 amino acid residues with a molecular weight of approximately 15 kDa, containing three cysteine ​​residues located at positions 58, 105, and 125. Post-translational modifications include Thr glycosylation at position 3, and the formation of disulfide bonds between the cysteine ​​residues at positions 58 and 105, resulting in its essential higher-order structure, primarily composed of four α-helices and several loops (Bazan et al., Science 257, 410-413 (1992)).

[0003] IL-2 is mainly produced by activated T cells. It can promote the proliferation and differentiation of T cells and maintain T cell activity; stimulate the generation, proliferation and activation of natural killer (NK) cells, and induce the generation of cytotoxic T lymphocytes (CTLs) as well as induce and activate lymphokine-activated killer (LAK) cells and tumor-infiltrating lymphocytes; promote the expression of cytokines and cell lysis molecules by T cells, and promote the proliferation of B cells (Waldmann et al., Nat Rev Immunol 6, 595-601 (2009)). These cells have the effect of killing cells infected by exogenous microorganisms and cancerous cells, so IL-2 has good antiviral and anticancer effects and broad clinical application potential.

[0004] IL-2 mediates its action by binding to the IL-2 receptor (IL-2R), which consists of three subunits: α (CD25), β (CD122), and γ (CD132). The α receptor is primarily expressed on T suppressor cells (T cells). reg It is expressed on the surface of some endothelial cells and T cells, while the β and γ receptor subunits are highly expressed on effector T cells (T cells). effIL-2 has different affinities for complexes of different receptor subunits. IL-2 has the highest affinity for complexes composed of α, β, and γ receptor subunits, and a moderate affinity (approximately 100-fold lower) for complexes composed of β and γ receptor subunits. IL-2 can transmit signals after binding to both types of receptor subunits (Minami et al., Annu RevImmunol 11, 245-268 (1993)). However, in clinical settings, low doses of IL-2 preferentially bind to high-affinity receptors on the surface of Treg cells, leading to immunosuppression and failing to achieve the therapeutic effect. High doses of IL-2 can neutralize the immunosuppression caused by Treg activation by activating a large number of effector T cells, but also result in more toxic side effects and activation-induced cell apoptosis.

[0005] Based on the antitumor effects of IL-2, high-dose IL-2 (aldeleukin) was approved by the FDA in 1992 for the clinical treatment of melanoma and renal cell carcinoma. However, patients receiving high-dose IL-2 treatment often experience serious side effects, including cardiovascular, pulmonary edema, hepatic, gastrointestinal, neurological, and hematological events. Most of these side effects can be explained by vascular (or capillary) leakage syndrome (VLS), which is also an indicator for evaluating the side effects of IL-2 treatment in clinical and animal experiments. VLS is caused by the expression of high-affinity receptors (α, β, and γ subunits) of IL-2 on endothelial cells (Krieg et al., Proc Nat Acad Sci USA 107, 11906-11 (2010)). Therefore, weakening or eliminating the binding to α receptors will help reduce the function of IL-2 in promoting the inhibition of T cell proliferation; at the same time, it can also reduce the binding to endothelial cell α receptors, thereby reducing or eliminating the toxic side effects caused by IL-2 treatment. The binding sites of IL-2 to the α receptor subunit are mainly at amino acid positions 37, 38, 41, 42, 43, 44, 45, 61, 62, 65, 68, and 72 (Rickert.M et al. (2005) Science 308:1477-1480). Merck, Roche, and other research institutions have made some mutations around these IL-2 surface amino acids that bind to the α receptor subunit. For example, Merck's mutants (R38W, F42K, WO2008003473A2) reduce the interaction with the α receptor subunit, thereby achieving effector T cell activation and enhancing efficacy. Roche's IL-2 mutants (F42A, Y45A, and L72G, US 2016 / 0208017A1) do not bind to the α receptor, but can bind normally to the β and γ receptor subunit complex and exert their effects. They are currently being used in clinical trials.

[0006] Therefore, reducing or eliminating the interaction between IL-2 and the α receptor subunit may be an important aspect of treatment effectiveness and reducing treatment side effects in cancer patients. Summary of the Invention

[0007] In view of the deficiencies in the prior art, the present invention provides an IL-2 derivative and its complex.

[0008] One aspect of the present invention provides an IL-2 derivative, which, in one specific embodiment, partially or completely blocks the binding plane of the IL-2 derivative to the α receptor subunit by introducing at least one cysteine ​​residue on the basis of IL-2, while substantially retaining the affinity for the β and γ receptor subunit complex.

[0009] Furthermore, the introduction of at least one cysteine ​​residue into IL-2 enables:

[0010] a) Formation of intramolecular disulfide bonds in IL-2 derivatives; or

[0011] b) Enables the IL-2 derivative to bind to the closed module via intermolecular disulfide bonds.

[0012] Optionally, the introduction of at least one cysteine ​​residue is achieved by introducing at least one cysteine ​​residue through point mutation.

[0013] Optionally, the IL-2 is wild-type IL-2, whose amino acid sequence is shown in SEQ ID NO.1.

[0014] Optionally, in one approach, a first cysteine ​​residue and a second cysteine ​​residue are introduced into IL-2 via point mutation; one or both of the first and second cysteine ​​residues are amino acids or nearby amino acids associated with the binding plane of wild-type IL-2 to the α-receptor subunit. Specifically, the amino acid sites associated with the binding plane of wild-type IL-2 to the α-receptor subunit are positions 37, 38, 41, 42, 43, 44, 45, 61, 62, 65, 68, and 72.

[0015] Further, the first cysteine ​​residue is amino acid 37, 38, 41, 42, 43, 44, 45, 61, or 62 of wild-type IL-2, or an amino acid nearby; the second cysteine ​​residue is amino acid 61, 62, 65, 68, or 72 of wild-type IL-2, or an amino acid nearby. Optionally, "nearby" means: 1) 1 to 4 amino acids adjacent in the primary structure; and / or amino acids adjacent in the tertiary structure.

[0016] Furthermore, the first cysteine ​​residue is an amino acid point mutation selected from the group consisting of: K35C, L36C, R38C, M39C, L40C, T41C, F42C, K43C, F44C, and E61C.

[0017] Furthermore, the second cysteine ​​residue is an amino acid point mutation selected from the group consisting of: V69C, E62C, P65C, T111C, Y107C, A112C, T113C, I114C, L72C, and A73C.

[0018] Optionally, the combination of the first and second cysteine ​​residues forming the intramolecular disulfide bond is selected from the following amino acid point mutation combinations: M39C and V69C, F44C and E62C, F44C and P65C, F42C and V69C, E61C and Y107C, F42C and P65C, F42C and T111C, F42C and A112C, F42C and T11 3C, T41C and A112C, L40C and A112C, L40C and T113C, L40C and I114C, M39C and L72C, M39C and A73C, R38C and V69C, R38C and L72C, L36C and V69C, L36C and L72C, L36C and A73C, K35C and V69C, and K43C and A112C.

[0019] Optionally, the centroid vector distance between the first cysteine ​​residue and the second cysteine ​​residue is less than [missing value].

[0020] Alternatively, in the second approach, a third cysteine ​​residue is introduced into IL-2 via point mutation; the third cysteine ​​residue is an amino acid associated with the binding plane of IL-2 to the α-receptor subunit or an amino acid in its vicinity.

[0021] Optionally, the IL-2 is wild-type IL-2, whose amino acid sequence is shown in SEQ ID NO.1.

[0022] Further, the third cysteine ​​residue is the amino acid at position 37, 38, 41, 42, 43, 44, 45, 61, 62, 65, 68, or 72 of wild-type IL-2, or an amino acid nearby. Optionally, "nearby" means: 1) 1 to 4 amino acids adjacent in the primary structure; and / or amino acids adjacent in the tertiary structure.

[0023] Furthermore, the third cysteine ​​residue is an amino acid point mutation selected from the group consisting of: P34C, K35C, T37C, R38C, T41C, K43C, F44C, Y45C, E61C, E62C, K64C, P65C, E68C, and L72C.

[0024] Furthermore, the closed module has or has a fourth cysteine ​​residue introduced therein; the third cysteine ​​residue on the IL-2 derivative can form an intermolecular disulfide bond with the fourth cysteine ​​residue on the closed module.

[0025] Optionally, the centroid vector distance between the third cysteine ​​residue and the fourth cysteine ​​residue is less than [value missing].

[0026] Optionally, the closed module is the extracellular segment of the α-receptor subunit.

[0027] Furthermore, the amino acid sequence of the extracellular segment of the α-receptor subunit is shown in SEQ ID NO.24.

[0028] Furthermore, the fourth cysteine ​​residue on the extracellular segment of the α-receptor subunit is selected from the following amino acid point mutations: D4C, D5C, M25C, N27C, R35C, R36C, K38C, S39C, G40C, S41C, L42C, I118C, Y119C, and H120C.

[0029] Optionally, the combination of the third and fourth cysteine ​​residues forming the intermolecular disulfide bond is selected from the following amino acid point mutation combinations: T41C and N27C, P34C and D4C, E68C and L42C, Y45C and R35C, R38C and H120C, L72C and M25C, E61C and S39C, T41C and I118C, K35C and D4C, T37C and D4C, R38C and D4C, R38C and D5C, T41C and L42C, T41C and Y119C, K43C and R35C, K43C and R36C, F44C and L42C, K43C and L42C, E61C and K38C, E62C and K38C, K64C and S39C, K64C and G40C, K64C and S41C, P65C and K38C.

[0030] Alternatively, in either the first or second method, the cysteine ​​residue at position 125 of wild-type IL-2 can be replaced with another amino acid residue via point mutation.

[0031] Alternatively, the point mutation at position 125 of wild-type IL-2 can be C125A.

[0032] Optionally, the amino acid sequence of the IL-2 derivative is shown in SEQ ID NO.3 to SEQ ID NO.24; or in SEQ ID NO.26 to 40.

[0033] A second aspect of the present invention provides a composite, in one specific embodiment, comprising:

[0034] 1) An IL-2 derivative, wherein a third cysteine ​​residue is introduced into IL-2;

[0035] 2) A closed module having or having a fourth cysteine ​​residue introduced thereon;

[0036] The third cysteine ​​residue on the IL-2 derivative and the fourth cysteine ​​residue on the blocking module form an intermolecular disulfide bond, thereby forming a complex of IL-2 and the blocking module, which partially or completely blocks the binding plane of the IL-2 derivative to the α receptor subunit, while essentially retaining the affinity for the complex of the β and γ receptor subunits.

[0037] Optionally, the introduction of at least one cysteine ​​residue is performed by introducing at least one cysteine ​​residue through point mutation.

[0038] Further, the third cysteine ​​residue is an amino acid associated with the binding plane of IL-2 to the α-receptor subunit or an amino acid nearby therefrom. Optionally, the IL-2 is wild-type IL-2, whose amino acid sequence is shown in SEQ ID NO.1.

[0039] Further, the third cysteine ​​residue is the amino acid at position 37, 38, 41, 42, 43, 44, 45, 61, 62, 65, 68, or 72 of wild-type IL-2, or an amino acid nearby. Optionally, "nearby" means: 1) 1 to 4 amino acids adjacent in the primary structure; and / or amino acids adjacent in the tertiary structure.

[0040] Optionally, the centroid vector distance between the third cysteine ​​residue and the fourth cysteine ​​residue is less than [missing value].

[0041] Furthermore, the third cysteine ​​residue is an amino acid point mutation selected from the following group: P34C, K35C, T37C, R38C, T41C, K43C, F44C, Y45C, E61C, E62C, K64C, P65C, E68C, and L72C.

[0042] Optionally, the closed module is an extracellular segment of the α-receptor subunit, the amino acid sequence of which is shown in SEQ ID NO.25.

[0043] Furthermore, the fourth cysteine ​​residue on the extracellular segment of the α-receptor subunit is selected from the following amino acid point mutations: D4C, D5C, M25C, N27C, R35C, R36C, K38C, S39C, G40C, S41C, L42C, I118C, Y119C, and H120C.

[0044] Optionally, the combination of the third and fourth cysteine ​​residues forming the intermolecular disulfide bond is selected from the following amino acid point mutation combinations: T41C and N27C, P34C and D4C, E68C and L42C, Y45C and R35C, R38C and H120C, L72C and M25C, E61C and S39C, T41C and I118C, K35C and D4C, T37C and D4C, R38C and D4C, R38C and D5C, T41C and L42C, T41C and Y119C, K43C and R35C, K43C and R36C, F44C and L42C, K43C and L42C, E61C and K38C, E62C and K38C, K64C and S39C, K64C and G40C, K64C and S41C, P65C and K38C.

[0045] Optionally, a point mutation at position 125 of wild-type IL-2 is performed as C125A.

[0046] Optionally, the combination of the amino acid sequence of the IL-2 derivative and the sequence of the extracellular segment of the α-receptor subunit is selected from the group consisting of: SEQ ID NO.26 and SEQ ID NO.50, SEQ ID NO.27 and SEQ ID NO.51, SEQ ID NO.28 and SEQ ID NO.52, SEQ ID NO.29 and SEQ ID NO.53, SEQ ID NO.30 and SEQ ID NO.54, SEQ ID NO.31 and SEQ ID NO.55, SEQ ID NO.32 and SEQ ID NO.56, SEQ ID NO.33 and SEQ ID NO.57, SEQ ID NO.34 and SEQ ID NO.58, SEQ ID NO.35 and SEQ ID NO.59, SEQ ID NO.36 and SEQ ID NO.60, SEQ ID NO.37 and SEQ ID NO.61, SEQ ID NO.38 and SEQ ID NO.62, SEQ ID NO.39 and SEQ ID NO.63, SEQ ID NO.40 and SEQ ID NO.64, SEQ ID NO. SEQ ID NO.41 and SEQ ID NO.65, SEQ ID NO.42 and SEQ ID NO.66, SEQ ID NO.43 and SEQ ID NO.67, SEQ ID NO.44 and SEQ ID NO.68, SEQ ID NO.45 and SEQ ID NO.69, SEQ ID NO.46 and SEQ ID NO.70, SEQ ID NO.47 and SEQ ID NO.71, SEQ ID NO.48 and SEQ ID NO.72, SEQ ID NO.49 and SEQ ID NO.73.

[0047] A third aspect of the invention provides an isolated polynucleotide, in one specific embodiment, which encodes an IL-2 derivative as described above, or a complex as described above.

[0048] A fourth aspect of the present invention provides an expression vector, in one specific embodiment, comprising a separated polynucleotide as described above.

[0049] A fifth aspect of the invention provides a host cell, in one specific embodiment, comprising a separated polynucleotide as described above.

[0050] A sixth aspect of the invention provides a composition, in one embodiment, comprising an IL-2 derivative or a complex as described above, and a pharmaceutically acceptable carrier.

[0051] A seventh aspect of the invention provides the use of the IL-2 derivatives or complexes described above in the preparation of medicaments or formulations for treating diseases.

[0052] The eighth aspect of the invention provides the use of the IL-2 derivatives or complexes described above in the preparation of compositions for stimulating the immune system of an individual.

[0053] A ninth aspect of the present invention provides a method for generating an IL-2 derivative, in one specific embodiment of which the method includes culturing a host cell as described above under conditions suitable for expressing the IL-2 derivative.

[0054] A tenth aspect of the invention provides a method for generating a complex, in one specific embodiment of which the method includes culturing a host cell as described in claim 37 under conditions suitable for expressing the complex.

[0055] In specific embodiments of the present invention, the IL-2 derivatives or complexes, by forming intramolecular or intermolecular disulfide bonds, block the binding site of the IL-2 derivative with the α-receptor, thereby preventing binding to the α-receptor structure. The IL-2 derivatives or complexes of the present invention offer a new direction for reducing VLS or lowering or eliminating the toxic side effects caused by IL-2 therapy. Attached Figure Description

[0056] Figure 1 This is an example of SDS-PAGE electrophoresis of purified IL-2 derivatives in one embodiment of the present invention; wherein, reduction is achieved by adding the reducing agent β-ME to the loading buffer, and non-reduction is achieved by not adding the reducing agent.

[0057] Figure 2 This is a signal graph from one embodiment of the present invention, obtained by Fortebio testing the binding ability of an IL-2 derivative to IL2Rα at a concentration of 100 nM; the unrelated protein control used is HER2. Figure 2 A is for IL-2wt C125A; Figure 2 B targets IL-2 mutant 1; Figure 2 C targets IL-2 mutant 2; Figure 2 D targets IL-2 mutant 3; Figure 2 E targets IL-2 mutant 4; Figure 2 F targets IL-2 complex 1; Figure 2 G targets IL-2 complex 2; Figure 2 H targets IL-2 complex 3; Figure 2 I targets IL-2 complex 4.

[0058] Figure 3This is a Fortebio assay signal graph, along with Ka, kd, KD, and R2, from one embodiment of the present invention, to determine the binding affinity of an IL-2 derivative with IL-2Rβγ. The concentration range is 1.25 nM–40 nM. Figure 3 A is for IL-2wt C125A; Figure 3 B targets IL-2 mutant 1; Figure 3 C targets IL-2 mutant 2; Figure 3 D targets IL-2 mutant 3; Figure 3 E targets IL-2 mutant 4; Figure 3 F targets IL-2 complex 1; Figure 3 G targets IL-2 complex 2; Figure 3 H targets IL-2 complex 3; Figure 3 I targets IL-2 complex 4.

[0059] Figure 4 This is a proliferation experiment of CTLL-2 (T cells) in one embodiment of the present invention. Detailed Implementation

[0060] The present invention will be further described below with reference to embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of protection of the present invention.

[0061] Unless otherwise specified, experimental methods in the following examples were performed under standard conditions, such as those described in Sambrook et al., *Molecular Cloning: A Laboratory Manual* (New York: Cold Spring Harbor Laboratory Press, 1989), or as recommended by the manufacturer. Unless otherwise specified, all reagents used were commercially available or publicly available.

[0062] In this article, the relevant amino acid positions of IL-2 derivatives and wild-type IL-2 were calculated based on the amino acid sequence of wild-type IL-2 (such as SEQ ID NO.1).

[0063] Wild-type IL-2 (IL-2wt, SEQ ID NO.1):

[0064] APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHLPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLT.

[0065] The nucleotide sequence of wild-type IL-2 is shown in SEQ ID NO.146.

[0066] In this paper, the relevant amino acid positions of CD25-ECD were calculated based on the amino acid sequence shown in SEQ ID NO.25.

[0067] In this article, "first," "second," "third," etc., are used only for distinction and are not a requirement of order. For example, a derivative may contain only the "third" cysteine ​​residue, without necessarily having the "first" and "second" cysteine ​​residues.

[0068] In this article, "IL-2 derivatives" include IL-2 mutants and complexes formed by IL-2 mutants and other molecules. An IL-2 mutant is a molecule formed by mutation (such as a point mutation or insertion mutation) in IL-2. Optionally, the IL-2 is wild-type IL-2.

[0069] In this article, "IL2Rα" refers to interleukin-2 receptor α, also known as "α receptor subunit"; "IL2Rβ" refers to interleukin-2 receptor β, also known as "β receptor subunit"; "IL2Rγ" refers to interleukin-2 receptor γ, also known as "γ receptor subunit"; and "IL2Rβγ" refers to the complex formed by interleukin-2 receptor β and receptor γ, also known as "β and γ receptor subunit complex".

[0070] In existing technologies, in order to reduce or eliminate the binding of IL-2 to the α receptor, simple mutations are made to the amino acids on the binding surface of IL-2 to the α receptor.

[0071] In a specific embodiment of the present invention, the strategy adopted is to introduce additional cysteine ​​residues to form new disulfide bonds inside the IL-2 molecule, or to make IL-2 form a complex with other blocking modules through intermolecular disulfide bonds, thereby partially or completely blocking the binding site of IL-2 with the α receptor, but without affecting the binding with the β and γ receptor subunit complex.

[0072] In the first specific embodiment, the IL-2 derivative is structurally more stable by introducing a first cysteine ​​residue and a second cysteine ​​residue to form an intramolecular disulfide bond, and can also form a barrier to disrupt the binding plane with the α receptor.

[0073] The introduction of the first and second cysteine ​​residues involves appropriate point mutations in IL-2. The positions of the first and second cysteine ​​residues to be introduced are determined primarily through the following methods:

[0074] 1) The positions of the first and second cysteine ​​residues are amino acid residues on or near the binding surface of IL-2 and the α receptor; the amino acid residues on the binding surface of IL-2 and the α receptor are amino acid sites at positions 37, 38, 41, 42, 43, 44, 45, 61, 62, 65, 68 and 72.

[0075] 2) The structure and interatomic distances of IL-2 were fully considered;

[0076] 3) Through bioinformatics and protein engineering design, two suitable sites are found around the above amino acid residues based on the vector distance criterion of the centroid of the residues. These sites can form intramolecular disulfide bonds, but are not likely to form intermolecular disulfide bonds of IL-2.

[0077] After identifying suitable sites, the original amino acid residues are mutated to first and second cysteine ​​residues. The mutated IL-2 derivatives (and IL-2 mutants) then form intramolecular disulfide bonds through normal transcription and translation.

[0078] In some embodiments, the original free cysteine ​​residue at position 125 of IL-2 is mutated to prevent it from interfering with the formation of intermolecular disulfide bonds.

[0079] In some embodiments, the amino acid sequences of the designed IL-2 mutants are shown in Table 1:

[0080] Table 1. Amino acid sequences of the IL-2 mutants obtained through design.

[0081]

[0082]

[0083]

[0084] The expression host can be E. coli or mammalian cells.

[0085] In the second specific embodiment, IL-2 forms a complex with the blocking module via intermolecular disulfide bonds, thereby completely or partially blocking the binding plane of IL-2 to the α receptor, and thus blocking the binding of IL-2 to the endogenous α receptor. The disulfide bond between IL-2 and the blocking module is formed by introducing a third cysteine ​​residue onto IL-2 and having or introducing a fourth cysteine ​​residue onto the blocking module, and is formed between the third and fourth cysteine ​​residues.

[0086] In one embodiment, the blocking module is the extracellular segment of the α receptor. The amino acid sequence of the extracellular segment of the wild-type α receptor is shown in SEQ ID NO. 25. In its native state, the binding of the α receptor to IL-2 is unstable (high dissociation coefficient Kd) and cannot form a stable heterodimer. It must co-form a high-affinity receptor for IL-2 with the β and γ receptor subunits. Therefore, a stable complex cannot be formed by co-expressing two wild-type molecules. However, by introducing a third cysteine ​​residue onto IL-2 and a fourth cysteine ​​residue onto the extracellular segment (CD25-ECD) of the α receptor, a disulfide bond is formed between the third and fourth cysteine ​​residues, allowing the IL-2 derivative and the extracellular segment (CD25-ECD) of the α receptor to form a complex, thus blocking the binding of IL-2 to the endogenous α receptor.

[0087] The amino acid sequence of the extracellular segment of the wild-type α receptor is shown in SEQ ID NO.25: ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG;

[0088] The nucleotide sequence of the extracellular segment of the wild-type α receptor is shown in SEQ ID NO.145.

[0089] The positions of the third and fourth cysteine ​​residues to be introduced are mainly determined in the following ways:

[0090] 1) The position of the third cysteine ​​residue is an amino acid residue on or near the binding surface of IL-2 and the α receptor; the amino acid residues on the binding surface of IL-2 and the α receptor are amino acid sites at positions 37, 38, 41, 42, 43, 44, 45, 61, 62, 65, 68 and 72.

[0091] 2) Through bioinformatics and protein engineering design, suitable binding sites were found in the extracellular regions of IL-2 and α receptor (CD25-ECD) based on the vector distance criterion of the centroid of residues. This mutation does not affect the structure of the protein.

[0092] After identifying suitable sites, mutations were performed to change the original amino acid residues on IL-2 to a third cysteine ​​residue, and to change the original amino acid residues on the extracellular domain of the α-receptor (CD25-ECD) to a fourth cysteine ​​residue. Following co-expression of the mutants of IL-2 and the α-receptor extracellular domain, a disulfide bond was formed between IL-2 and the α-receptor extracellular domain after transcription and translation, resulting in a novel molecule, the IL-2 / CD25-ECD heterodimer. This complex cannot bind to endogenous α-receptors in vivo, but it can bind to the β and γ receptor subunit complexes, thereby preventing Treg activation.

[0093] In one embodiment, the original free cysteine ​​residue at position 125 of IL-2 is mutated to prevent it from forming additional disulfide bonds with the extracellular segment of the α receptor with the cysteine ​​mutation and IL-2, thereby affecting the formation of the α receptor-IL-2 dimer.

[0094] In some embodiments, the amino acid sequences of the designed IL-2 mutant and CD25-ECD mutant are shown in Table 2:

[0095] Table 2. Amino acid sequences of the IL-2 mutant and CD25-ECD mutant obtained by design.

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105] The host cells for expression are mammalian cells (HEK293 or CHO).

[0106] Example 1: Preparation of IL-2 (C125A), IL-2 mutant, and IL-2 complex

[0107] In this embodiment, IL-2wt(C125A), IL-2 mutants 1-4 and IL-2 complexes 1-4 were selected for expression and purified and prepared using the HPC4 tag at the C-terminus of the molecules.

[0108] 1.1 Construction of expression plasmids

[0109] Suzhou Genewise Biotechnology Co., Ltd. was commissioned to synthesize genes containing IL-2wt C125A (SEQ ID NO.74), IL-2 mutants 1-4, and IL-2 complexes 1-4 (IL-2Pair 1-4 and CD25-ECD Pair 1-4 in the IL-2 complex). Then, or according to the operation method mentioned in Molecular Cloning, overlap PCR was performed to obtain the target fragment. Then, the fragments and pTT5 universal vectors are recombined and ligated, transformed into DH10B, sequenced, and preserved to obtain the required IL-2wt(C125A), IL-2 mutants 1-4, and IL-2 complexes 1-4 plasmids (IL-2Pair 1-4 plasmids and CD25-ECDPair 1-4 plasmids in the IL-2 complex); or, following the procedures described in the Agilent QuikChange Lightning Site-Directed Mutagenesis Kit, PCR, DpnI digestion, transformation into DH10B, sequencing, and preservation are performed to obtain the required IL-2wt(C125A), IL-2 mutants 1-4, and IL-2 complexes 1-4 plasmids.

[0110] 1.2 Plasmid extraction and HEK293 cell preparation

[0111] 1.2.1 Plasmid Extraction

[0112] Following the procedures described in the Qiagen Mini-prep Kit and the Qiagen Endofree Maxi-prep Kit, plasmids for IL-2wt(C125A), IL-2 mutants 1-4, and IL-2 complexes 1-4 were prepared.

[0113] 1.2.2 HEK293 Cell Preparation

[0114] Freshly passaged HEK293 cells (National Research Council, Canada) at a density of 1–1.2 × 10^6 / ml were used for transient expression.

[0115] 1.3 Transient Expression of HEK293

[0116] 1.3.1 Reagent Preparation

[0117] A) G418 solution: Weigh 250mg of Geneticin TM Dissolve in 4.5ml of ultrapure water, bring the volume up to 5ml, filter through a 0.22um filter membrane, and store at -20℃.

[0118] B) PEI solution: Weigh 50mg PEI and dissolve it in 45ml of ultrapure water. Adjust the pH to 7.0 with 1M NaOH, bring the volume to 50ml with ultrapure water, filter through a 0.22um filter membrane, and store at -20℃.

[0119] C) Culture medium: In 1L FreeStyle TM Add 10 ml of Pluronic to 293 Expression Medium TM F-68 and 500ul G418;

[0120] D) Prepare the plasmid in advance in a 2ml endotoxin-free centrifuge tube;

[0121] E) Prepare a fresh cell suspension of 1-1.2 × 10⁶ cells / ml according to the volume required for transfection.

[0122] 1.3.2 Preparation of transfection reagent-plasmid complex

[0123] Solution A: Plasmid 1ug / ml + Opti-MEM™ 33.3ul / ml

[0124] Solution B: PEI 2ug / ml+Opti-MEMTM 33.3ul / ml

[0125] Pour solution B into solution A, mix well, and incubate for 10 minutes. Then add the cell suspension.

[0126] During transfection, the plasmids of IL-2wt(C125A) and IL-2 mutants 1-4 were transfected separately; the plasmids of IL-2 complexes 1-4 were transfected by mixing the two plasmids.

[0127] 1.3.3 Fluid Change

[0128] After incubation at 115 rpm, 36.8℃, and 5% CO2 for 4 h, centrifugation at 800 g for 5 min was performed, followed by replacement with FreeStyle without added F68 and G418. TM 293 Expression Medium

[0129] 1.3.4 Cultivation and Gains of Expression

[0130] After culturing at 115 rpm, 36.8℃, and 5% CO2 for 5 days, the cells were centrifuged at 8500 rpm for 15 min to collect the cell supernatant.

[0131] 1.4 Purification and Preparation

[0132] All IL-2-related derivatives have an HPC4 tag at their C-terminus, so affinity purification can be performed using a packing material conjugated with an HPC4 antibody, followed by further purification via gel filtration chromatography (Superdex 200) to obtain highly pure proteins. SDS-PAGE analysis was performed according to the method described in *Molecular Cloning*.

[0133] The results are as follows Figure 1 As shown, after the plasmids were transfected into the cells, IL-2wt(C125A), IL-2 mutants 1-4 and IL-2 complexes 1-4 were generated, and the plasmid construction, protein expression and purification were successful.

[0134] Example 2: The affinity of IL-2wt(C125A), IL-2 mutants 1-4, and IL-2 complexes 1-4 for IL-2Rβγ and IL-2Rα was determined by biolayer interferomeory (BLI).

[0135] 1. Experimental materials

[0136] All proteins used in the experiments were produced by Beijing Zhidao Biotechnology Co., Ltd. IL2Rα-his (purchased from Beijing Zhidao Biotechnology Co., Ltd.), IL2Rβγ-Fc (purchased from Beijing Zhidao Biotechnology Co., Ltd.), and the IL2 mutant were obtained through transient expression on HEK293 followed by affinity purification. Buffer formulation: 10 mM HEPES, 150 mM sodium chloride, 3 mM EDTA, 0.1% BSA, and 0.05% Tween 20; ProA sensor (Pall Fortebio, catalog number #18-5010); HISIK sensor (Pall Fortebio, catalog number #18-5120); BLI device: Octet RED96 (Pall Fortebio); Data acquisition and analysis were performed using Data Acquisition 11.0 and Data Analysis 11.0 software, respectively.

[0137] 2. Experimental Methods

[0138] 1) Preparation of IL2Rβγ-Fc

[0139] Dilute IL2Rβγ-Fc to a concentration of 10ug / ml with buffer, add it to column 2 of a 96-well assay plate, and set the control program to Loading for 600s.

[0140] 2) IL2Rα-his preparation

[0141] Dilute IL2Rα-his with buffer to a concentration of 10 μg / ml, add it to column 3 of a 96-well assay plate, and set the control program to Loading for 600 seconds.

[0142] 3) Sample preparation

[0143] The IL-2 derivative was diluted to 100 nM with buffer, and then serially diluted 1:1 down to 6 gradients (7 gradients in total) to a concentration of 1.625 nM and 0. These were then added to columns 5-9 of a 96-well plate, with the program set to Association and running for 200 seconds. Buffer was added to columns 1, 4, 10, and 11 of the 96-well plate, and glycine at pH 1.7 was added to column 12. The volume of both the sample and solution added was 200 μL.

[0144] 4) Detect the affinity with IL2Rβγ-Fc

[0145] Eight ProA sensors were placed in the AH slot of the first column of the sensor holder. The detection conditions were set in Data Acquisition 11.0 software as follows: 1. Pre-humidification: Baseline, 60s, Position: Column 1. 2. Cyclic detection: Column 2: Loading, 600s; Column 4: Baseline 1, 60s; Samples: Columns 5-9: Association, 200s; Column 10: Dissociation, 600s; Column 11: Neutralization; Column 12: Regeneration.

[0146] 5) Detection of binding with IL2Rα

[0147] Eight HISIK sensors were placed in the AH slot of the second column of the sensor holder. The detection conditions were set in Data Acquisition 11.0 software as follows: 1. Pre-humidification: Baseline, 60s, Position: Column 1. 2. Cyclic detection: Column 3: Loading, 600s; Column 4: Baseline 1, 60s; Samples: Columns 5-9: Association, 200s; Column 10: Dissociation, 600s; Column 11: Neutralization; Column 12: Regeneration.

[0148] 6) Data Analysis

[0149] Data was analyzed using Data Analysis 11.0 software. With 0 concentration as the control and background subtraction, KD values ​​were calculated using a Fitting curve.

[0150] 3. Results

[0151] like Figure 2As shown, from the binding curves of IL-2 derivatives and the IL-2Rα receptor, except for IL-2 mutant 1 which significantly weakened its binding to the IL-2Rα receptor, the other IL-2 derivatives almost did not bind to it; for example... Figure 3 As shown, the binding curves of IL-2 derivatives to the IL-2Rβγ receptor show no significant changes compared to IL-2wt C125A. Figure 3 ).

[0152] Example 3: Experiment to promote T cell proliferation

[0153] The CTLL-2 (T cell) proliferation assay is a commonly used cellular-level assay to determine the activity of interleukin-stimulated immune cells. Therefore, this study uses a CTLL-2 cell proliferation assay to examine the biological activity of IL-2 derivatives.

[0154] 1) CTLL-2 cell preparation: Resuspend the cells in a culture medium containing FBS and Rat-T-Stim.

[0155] 2) Sample loading: Seed cells into 96-well plates, 0.1 ml per well. Simultaneously, serially dilute the IL-2 derivatives 11, 18, 21, and 28 (i.e., the proteins prepared in Example 1), adding 0.1 ml to each well, with three replicates for each dilution. A control well (100 μL cells + 100 μL culture medium) was also included. Incubate at 37°C, 5% CO2 for 72 hours.

[0156] 3) MTS addition: Add 20 μl to each well. AQueous One Solution Reagent, incubate at 37 degrees Celsius with 5% CO2 for 2–4 hours.

[0157] 4) Measurement: Use an enzyme-linked immunosorbent assay (ELISA) reader to measure the absorbance (A) at a wavelength of 490 nm and calculate the EC50 value.

[0158] 5) Results: The selected representative IL-2 complex 2 and IL-2 mutant 4 both possessed CTLL-2 (T cell) proliferative activity, indicating that they did not significantly affect the signal transduction function of the β and γ receptor subunit complex. Figure 4 ). SEQUENCE LISTING <110> Beijing Zhidao Biotechnology Co., Ltd. <120> Interleukin-2 derivatives <160> 146 <170> PatentIn version 3.5 <210> 1 <211> 133 <212> PRT <213> Homo sapiens <400> 1 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 2 <211> 133 <212> PRT <213> Artificial Sequence <400> 2 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 3 <211> 133 <212> PRT <213> Artificial Sequence <400> 3 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Cys Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Cys Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 4 <211> 133 <212> PRT <213> Artificial Sequence <400> 4 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Cys Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Cys Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 5 <211> 133 <212> PRT <213> Artificial Sequence <400> 5 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Cys Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Cys Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 6 <211> 133 <212> PRT <213> Artificial Sequence <400> 6 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Cys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Cys Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 7 <211> 133 <212> PRT <213> Artificial Sequence <400> 7 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Cys Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Cys Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 8 <211> 133 <212> PRT <213> Artificial Sequence <400> 8 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Cys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Cys Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 9 <211> 133 <212> PRT <213> Artificial Sequence <400> 9 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Cys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Cys Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 10 <211> 133 <212> PRT <213> Artificial Sequence <400> 10 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Cys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Cys 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 11 <211> 133 <212> PRT <213> Artificial Sequence <400> 11 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Cys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Cys Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 12 <211> 133 <212> PRT <213> Artificial Sequence <400> 12 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Cys Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Cys 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 13 <211> 133 <212> PRT <213> Artificial Sequence <400> 13 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Cys Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Cys 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 14 <211> 133 <212> PRT <213> Artificial Sequence <400> 14 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Cys Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Cys Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 15 <211> 133 <212> PRT <213> Artificial Sequence <400> 15 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Cys Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Cys Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 16 <211> 133 <212> PRT <213> Artificial Sequence <400> 16 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Cys Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Cys Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 17 <211> 133 <212> PRT <213> Artificial Sequence <400> 17 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Cys Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Cys Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 18 <211> 133 <212> PRT <213> Artificial Sequence <400> 18 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Cys Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 19 <211> 133 <212> PRT <213> Artificial Sequence <400> 19 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Cys Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 20 <211> 133 <212> PRT <213> Artificial Sequence <400> 20 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Cys Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Cys Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 21 <211> 133 <212> PRT <213> Artificial Sequence <400> 21 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Cys Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Cys Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 22 <211> 133 <212> PRT <213> Artificial Sequence <400> 22 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Cys Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Cys Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 23 <211> 133 <212> PRT <213> Artificial Sequence <400> 23 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Cys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Cys Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 24 <211> 133 <212> PRT <213> Artificial Sequence <400> 24 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Cys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Cys 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 25 <211> 165 <212> PRT <213> Artificial Sequence <400> 25 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 26 <211> 133 <212> PRT <213> Artificial Sequence <400> 26 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Cys Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 27 <211> 133 <212> PRT <213> Artificial Sequence <400> 27 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Cys Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 28 <211> 133 <212> PRT <213> Artificial Sequence <400> 28 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Cys Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 29 <211> 133 <212> PRT <213> Artificial Sequence <400> 29 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Cys Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 30 <211> 133 <212> PRT <213> Artificial Sequence <400> 30 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 31 <211> 133 <212> PRT <213> Artificial Sequence <400> 31 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Cys Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 32 <211> 133 <212> PRT <213> Artificial Sequence <400> 32 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Cys Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 33 <211> 133 <212> PRT <213> Artificial Sequence <400> 33 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Cys Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 34 <211> 133 <212> PRT <213> Artificial Sequence <400> 34 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Cys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 35 <211> 133 <212> PRT <213> Artificial Sequence <400> 35 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Cys Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 36 <211> 133 <212> PRT <213> Artificial Sequence <400> 36 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 37 <211> 133 <212> PRT <213> Artificial Sequence <400> 37 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 38 <211> 133 <212> PRT <213> Artificial Sequence <400> 38 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Cys Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 39 <211> 133 <212> PRT <213> Artificial Sequence <400> 39 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Cys Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 40 <211> 133 <212> PRT <213> Artificial Sequence <400> 40 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Cys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 41 <211> 133 <212> PRT <213> Artificial Sequence <400> 41 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Cys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 42 <211> 133 <212> PRT <213> Artificial Sequence <400> 42 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Cys Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 43 <211> 133 <212> PRT <213> Artificial Sequence <400> 43 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Cys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 44 <211> 133 <212> PRT <213> Artificial Sequence <400> 44 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Cys Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 45 <211> 133 <212> PRT <213> Artificial Sequence <400> 45 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Cys Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 46 <211> 133 <212> PRT <213> Artificial Sequence <400> 46 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Cys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 47 <211> 133 <212> PRT <213> Artificial Sequence <400> 47 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Cys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 48 <211> 133 <212> PRT <213> Artificial Sequence <400> 48 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Cys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 49 <211> 133 <212> PRT <213> Artificial Sequence <400> 49 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Cys Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 50 <211> 165 <212> PRT <213> Artificial Sequence <400> 50 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Cys Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 51 <211> 165 <212> PRT <213> Artificial Sequence <400> 51 Glu Leu Cys Cys Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 52 <211> 165 <212> PRT <213> Artificial Sequence <400> 52 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Cys Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 53 <211> 165 <212> PRT <213> Artificial Sequence <400> 53 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Cys Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 54 <211> 165 <212> PRT <213> Artificial Sequence <400> 54 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr Cys Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 55 <211> 165 <212> PRT <213> Artificial Sequence <400> 55 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Cys Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 56 <211> 165 <212> PRT <213> Artificial Sequence <400> 56 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Cys Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 57 <211> 165 <212> PRT <213> Artificial Sequence <400> 57 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Cys Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 58 <211> 165 <212> PRT <213> Artificial Sequence <400> 58 Glu Leu Cys Cys Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 59 <211> 165 <212> PRT <213> Artificial Sequence <400> 59 Glu Leu Cys Cys Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 60 <211> 165 <212> PRT <213> Artificial Sequence <400> 60 Glu Leu Cys Cys Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 61 <211> 165 <212> PRT <213> Artificial Sequence <400> 61 Glu Leu Cys Asp Cys Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 62 <211> 165 <212> PRT <213> Artificial Sequence <400> 62 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Cys Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 63 <211> 165 <212> PRT <213> Artificial Sequence <400> 63 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Cys His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 64 <211> 165 <212> PRT <213> Artificial Sequence <400> 64 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Cys Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 65 <211> 165 <212> PRT <213> Artificial Sequence <400> 65 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Cys Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 66 <211> 165 <212> PRT <213> Artificial Sequence <400> 66 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Cys Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 67 <211> 165 <212> PRT <213> Artificial Sequence <400> 67 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Cys Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 68 <211> 165 <212> PRT <213> Artificial Sequence <400> 68 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Cys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 69 <211> 165 <212> PRT <213> Artificial Sequence <400> 69 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Cys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 70 <211> 165 <212> PRT <213> Artificial Sequence <400> 70 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Cys Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 71 <211> 165 <212> PRT <213> Artificial Sequence <400> 71 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Cys Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 72 <211> 165 <212> PRT <213> Artificial Sequence <400> 72 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Cys Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 73 <211> 165 <212> PRT <213> Artificial Sequence <400> 73 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Cys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly 165 <210> 74 <211> 399 <212> DNA <213> Artificial Sequence <400> 74 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 75 <211> 399 <212> DNA <213> Artificial Sequence <400> 75 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagatgcctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agagtgtctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 76 <211> 399 <212> DNA <213> Artificial Sequence <400> 76 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt gctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gagtgtctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 77 <211> 399 <212> DNA <213> Artificial Sequence <400> 77 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt gctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agtgtctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 78 <211> 399 <212> DNA <213> Artificial Sequence <400> 78 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 acctgcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agagtgtctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 79 <211> 399 <212> DNA <213> Artificial Sequence <400> 79 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 tgcgagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagtg tgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 80 <211> 399 <212> DNA <213> Artificial Sequence <400> 80 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 acctgcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agtgtctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 81 <211> 399 <212> DNA <213> Artificial Sequence <400> 81 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 acctgcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag tgtgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 82 <211> 399 <212> DNA <213> Artificial Sequence <400> 82 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 acctgcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag acctgtacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 83 <211> 399 <212> DNA <213> Artificial Sequence <400> 83 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 acctgcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgcctgta tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 84 <211> 399 <212> DNA <213> Artificial Sequence <400> 84 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 tgcttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag acctgtacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 85 <211> 399 <212> DNA <213> Artificial Sequence <400> 85 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgtgc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag acctgtacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 86 <211> 399 <212> DNA <213> Artificial Sequence <400> 86 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgtgc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgcctgta tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 87 <211> 399 <212> DNA <213> Artificial Sequence <400> 87 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgtgc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacct gtgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 88 <211> 399 <212> DNA <213> Artificial Sequence <400> 88 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagatgcctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aattgtgccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 89 <211> 399 <212> DNA <213> Artificial Sequence <400> 89 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagatgcctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctgtgtc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 90 <211> 399 <212> DNA <213> Artificial Sequence <400> 90 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac atgcatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agagtgtctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 91 <211> 399 <212> DNA <213> Artificial Sequence <400> 91 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac atgcatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aattgtgccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 92 <211> 399 <212> DNA <213> Artificial Sequence <400> 92 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagtgcac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agagtgtctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 93 <211> 399 <212> DNA <213> Artificial Sequence <400> 93 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagtgcac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aattgtgccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 94 <211> 399 <212> DNA <213> Artificial Sequence <400> 94 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagtgcac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctgtgtc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 95 <211> 399 <212> DNA <213> Artificial Sequence <400> 95 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc cctgcctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agagtgtctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 96 <211> 399 <212> DNA <213> Artificial Sequence <400> 96 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttctgct tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag acctgtacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 97 <211> 399 <212> DNA <213> Artificial Sequence <400> 97 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 tgcttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 98 <211> 399 <212> DNA <213> Artificial Sequence <400> 98 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatt gcaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 99 <211> 399 <212> DNA <213> Artificial Sequence <400> 99 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga atgcgtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 100 <211> 399 <212> DNA <213> Artificial Sequence <400> 100 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctgcatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 101 <211> 399 <212> DNA <213> Artificial Sequence <400> 101 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac atgcatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 102 <211> 399 <212> DNA <213> Artificial Sequence <400> 102 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aattgcgccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 103 <211> 399 <212> DNA <213> Artificial Sequence <400> 103 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 tgcgagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 104 <211> 399 <212> DNA <213> Artificial Sequence <400> 104 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 tgcttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 105 <211> 399 <212> DNA <213> Artificial Sequence <400> 105 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc cctgcctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 106 <211> 399 <212> DNA <213> Artificial Sequence <400> 106 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgtg cagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 107 <211> 399 <212> DNA <213> Artificial Sequence <400> 107 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac atgcatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 108 <211> 399 <212> DNA <213> Artificial Sequence <400> 108 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac atgcatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 109 <211> 399 <212> DNA <213> Artificial Sequence <400> 109 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 tgcttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 110 <211> 399 <212> DNA <213> Artificial Sequence <400> 110 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 tgcttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 111 <211> 399 <212> DNA <213> Artificial Sequence <400> 111 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttctgct tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 112 <211> 399 <212> DNA <213> Artificial Sequence <400> 112 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttctgct tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 113 <211> 399 <212> DNA <213> Artificial Sequence <400> 113 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt gctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 114 <211> 399 <212> DNA <213> Artificial Sequence <400> 114 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttctgct tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 115 <211> 399 <212> DNA <213> Artificial Sequence <400> 115 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 tgcgagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 116 <211> 399 <212> DNA <213> Artificial Sequence <400> 116 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gagtgcctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 117 <211> 399 <212> DNA <213> Artificial Sequence <400> 117 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctgt gccctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 118 <211> 399 <212> DNA <213> Artificial Sequence <400> 118 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctgt gccctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 119 <211> 399 <212> DNA <213> Artificial Sequence <400> 119 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctgt gccctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 120 <211> 399 <212> DNA <213> Artificial Sequence <400> 120 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agtgcctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tcgcccagag catcatcagc acactgacc 399 <210> 121 <211> 495 <212> DNA <213> Artificial Sequence <400> 121 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctctg ttgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 122 <211> 495 <212> DNA <213> Artificial Sequence <400> 122 gagctctgct gtgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 123 <211> 495 <212> DNA <213> Artificial Sequence <400> 123 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tcttgttaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 124 <211> 495 <212> DNA <213> Artificial Sequence <400> 124 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat tttgtaggat caagagcggc 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 125 <211> 495 <212> DNA <213> Artificial Sequence <400> 125 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctactgt 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 126 <211> 495 <212> DNA <213> Artificial Sequence <400> 126 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa cctgtctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 127 <211> 495 <212> DNA <213> Artificial Sequence <400> 127 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagtgtggc 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 128 <211> 495 <212> DNA <213> Artificial Sequence <400> 128 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gtgttaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 129 <211> 495 <212> DNA <213> Artificial Sequence <400> 129 gagctctgct gtgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 130 <211> 495 <212> DNA <213> Artificial Sequence <400> 130 gagctctgct gtgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 131 <211> 495 <212> DNA <213> Artificial Sequence <400> 131 gagctctgct gtgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 132 <211> 495 <212> DNA <213> Artificial Sequence <400> 132 gagctctgcg actgtgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 133 <211> 495 <212> DNA <213> Artificial Sequence <400> 133 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tcttgttaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 134 <211> 495 <212> DNA <213> Artificial Sequence <400> 134 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctgtcac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 135 <211> 495 <212> DNA <213> Artificial Sequence <400> 135 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat tttgtaggat caagagcggc 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 136 <211> 495 <212> DNA <213> Artificial Sequence <400> 136 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggtgtat caagagcggc 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 137 <211> 495 <212> DNA <213> Artificial Sequence <400> 137 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tcttgttaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 138 <211> 495 <212> DNA <213> Artificial Sequence <400> 138 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tcttgttaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 139 <211> 495 <212> DNA <213> Artificial Sequence <400> 139 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat ctgtagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 140 <211> 495 <212> DNA <213> Artificial Sequence <400> 140 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat ctgtagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 141 <211> 495 <212> DNA <213> Artificial Sequence <400> 141 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagtgtggc 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 142 <211> 495 <212> DNA <213> Artificial Sequence <400> 142 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagctgt 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 143 <211> 495 <212> DNA <213> Artificial Sequence <400> 143 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tgtctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 144 <211> 495 <212> DNA <213> Artificial Sequence <400> 144 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat ctgtagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 145 <211> 495 <212> DNA <213> Artificial Sequence <400> 145 gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60 aaaggggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcgg 120 tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180 tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240 aagcaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300 cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360 tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420 ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480 ctgatttgta ccgggg 495 <210> 146 <211> 399 <212> DNA <213> Artificial Sequence <400> 146 gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60 ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120 accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180 gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240 aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300 accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360 tggatcacct tctgccagag catcatcagc acactgacc 399

Claims

1. A complex possessing T cell proliferation activity, characterized in that, This complex is a heterodimer formed from a) an IL-2 derivative and b) a blocking module: a) An IL-2 derivative, wherein an additional third cysteine ​​residue is introduced by point mutation into wild-type IL-2 after a C125A mutation at position 125, and the amino acid sequence of the IL-2 derivative is shown in one of SEQ ID NO. 26 to SEQ ID NO. 29; b) A blocking module, wherein the blocking module is an extracellular CD25-ECD mutant of the α receptor, wherein a fourth cysteine ​​residue is introduced onto the extracellular CD25-ECD mutant of the α receptor, and the amino acid sequence of the blocking module is shown in one of SEQ ID NO. 50 to 53; An intermolecular disulfide bond is formed between the third and fourth cysteine ​​residues, causing the IL-2 derivative and the blocking module to form the complex; the IL-2 derivative forming the complex and the blocking module are selected from the following protein sequence combinations: SEQ ID NO. 26 and SEQ ID NO. 50, SEQ ID NO. 27 and SEQ ID NO. 51, SEQ ID NO. 28 and SEQ ID NO. 52, SEQ ID NO. 29 and SEQ ID NO.

53.

2. An isolated polynucleotide, characterized in that, Encoding the complex as described in claim 1.

3. An expression carrier, characterized in that, It contains the isolated polynucleotide as described in claim 2.

4. A host cell, characterized in that, It contains the isolated polynucleotide as described in claim 2.

5. A composition, characterized in that, It comprises the complex as described in claim 1, and a pharmaceutically acceptable carrier.

6. A method for generating the complex as described in claim 1, characterized in that, The method includes culturing the host cells as described in claim 4 under conditions suitable for expressing the complex.

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