PRE-FUSION-STABILIZED CMV GB PROTEIN NANOSTRUCTURES

Abstract

Provided herein are compositions and methods related to CMV gB proteins in which amino acid substitutions have been made to disrupt the postfusion state and/or stabilize the prefusion state.

Claims

1. A polypeptide, comprising an ectodomain of CMV gB in the prefusion conformation, wherein the ectodomain comprises, at positions relative to SEQ ID NO: 1: a. 1, 2, 3, 4 or more amino acid substitutions selected from the group consisting of E686L, R693V, V694L, Y690F, K700A, V702E, D679H, E681N, K695D, L680E, R685Q, V701L, E682S, F678N, N688E, V677T, D699K, F687A, M684S, Q692S, and Y696R, or substitutions at the same amino acid positions; b. the amino acid substitution S367I, the amino acid substitution T374F, or the amino acid substitutions S367I and T374F, or substitutions at positions 367 and/or 374; c. 1, 2, 3, 4 or more amino acid substitutions selected from the group consisting of Q591F, Q591Y, S668A, Y218F, N220S, V552L, or substitutions at the same amino acid positions; d, the amino acid substitution E167T; and/or e. 1, 2, 3, 4 or more amino acid substitutions selected from the group consisting of C246S, H157R, I156H, R457S, R460S, W240N, and Y242T.

2. The polypeptide of claim 1, wherein the ectodomain comprises the amino acid subsitutions D217C and S587C.

3. The polypeptide of claim 1, wherein the ectodomain comprises the amino acid subsitutions D217C and Y589C.

4. The polypeptide of any one of claims 1-3, wherein the ectodomain comprises 1, 2, 3, 4 or more amino acid substitutions selected from the group consisting of E686L, R693V, V694L, Y690F, K700A, V702E, D679H, E681N, K695D, L680E, R685Q, V701L, E682S, F678N, N688E, V677T, D699K, F687A, M684S, Q692S, and Y696R.

5. The polypeptide of claim 4, wherein the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, and Y690F.

6. The polypeptide of claim 4, wherein the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, K700A, and V702Q.

7. The polypeptide of claim 4, wherein the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, D679N, E681N, K695D, L680E, and R685Q.

8. The polypeptide of claim 4, wherein the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, K700A, V702Q, D679N, E681N, K695D, L680E, R685Q, and V701L.

9. The polypeptide of claim 4, wherein the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, D679N, E681N, K695D, L680E, R685Q, E682S, F678N, N688E, and V677T.

10. The polypeptide of claim 4, wherein the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, K700A, V702Q, D679N, E681N, K695D, L680E, R685Q, V701L, E682S, F678N, N688E, V677T, and D699K.

11. The polypeptide of claim 4, wherein the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, D679H, E681N, K695D, L680E, R685Q, E682S, F678N, N688E, V677T, F687A, M684S, Q692S, and Y696R.

12. The polypeptide of claim 4, wherein the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, K700A, V702E, D679H, E681N, K695D, L680E, R685Q, V701L, E682S, F678N, N688E, V677T, D699K, F687A, M684S, Q692S, and Y696R.

13. The polypeptide of any one of claims 1-12, wherein the ectodomain comprises the amino acid substitution S367I or the amino acid substitution T374F.

14. The polypeptide of any one of claims 1-12, wherein the ectodomain comprises the amino acid substitutions S367I and T374F.

15. The polypeptide of any one of claims 1-14, wherein the ectodomain comprises 1, 2, 3, 4 or more amino acid substitutions selected from the group consisting of Q591F, Q591Y, S668A, Y218F, N220S, and V552L.

16. The polypeptide of claim 15, wherein the ectodomain comprises the amino acid substitutions Q591F, S668A, and Y218F.

17. The polypeptide of claim 15, wherein the ectodomain comprises the amino acid substitutions N220S and V552L.

18. The polypeptide of claim 15, wherein the ectodomain comprises the amino acid substitutions Q591F, S668A, Y218F, N220S, and V552L.

19. The polypeptide of claim 15, wherein the ectodomain comprises the amino acid substitutions Q591Y, S668A, Y218F, N220S, and V552L.

20. The polypeptide of claim 15, wherein the ectodomain comprises the amino acid substitutions Q591Y, N220S, and V552L.

21. The polypeptide of claim 15, wherein the ectodomain comprises the amino acid substitutions Q591F, N220S, and V552L.

22. The polypeptide of any one of claims 1-21, wherein the ectodomain comprises amino acid substitution E167T or E167S.

23. The polypeptide of any one of claims 1-22, wherein the ectodomain comprises 1, 2, 3, 4 or more amino acid substitutions selected from the group consisting of C246S, H157R, I156H, R457S, R460S, W240N, and Y242T.

24. The polypeptide of claim any preceding claim, wherein the polypetide comprises any one combination of amino acid substitutions listed in Table 10.

25. The polypeptide of claim any preceding claim, wherein the polypetide comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a polypeptide sequence in Table 10 without the signal peptide, or an antigenic fragment thereof.

26. The polypeptide of claim 1, wherein the CMV gB protein adopts a prefusion conformation in the absence of a fusion inhibitor, optionally N-(4-[({(1S)-1-[3,5-bis(trifluoromethyl)phenyl|ethyl}carbamothioyl) amino]phenyl}-1,3-thiazole-4-carboxamide.

27. The polypeptide of any one of claims 1-23, wherein the polypeptide comprises, as a C-terminal fusion to the ectodomain, a trimerization domain.

28. The polypeptide of any one of claims 1-23, wherein the polypeptide comprises, as a C-terminal fusion to the ectodomain, a nanostructure assembly domain.

29. The polypeptide of claim 28, wherein the assembly domain comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to 153-50A (SEQ ID NO: 19), 153-50A.1 (SEQ ID NO: 21), 153-50A.1NegT2 (SEQ ID NO: 22), or 153-50A.1PosT1 (SEQ ID NO: 23).

30. The polypeptide of claim 28, wherein the assembly domain comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to 13-01 (SEQ ID NO: 8).

31. The polypeptide of claim 28, wherein the assembly domain comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, or SEQ ID NO: 31.

32. The polypeptide of claim 28, wherein the assembly domain comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to 153_dn5B (SEQ ID NO: 18).

33. A nanostructure comprising the polypeptide of any one of claim 29-32.

34. The nanostructure of claim 33, wherein the nanostructure comprises a second polypeptide.

35. The nanostructure of claim 33, wherein the second polypetpide comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to 153-50B (SEQ ID NO: 20), 153-50B.1 (SEQ ID NO: 24), 153-50B.1NcgT2 (SEQ ID NO: 25), or 153-50B.4PosT1 (SEQ ID NO: 26).

36. The nanostructure of claim 33, wherein the second polypeptide comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to 153_dn5A* (SEQ ID NO: 15), 153_dn5A.1 (SEQ ID NO: 16), or 153_dn5A.2 (SEQ ID NO: 17).

37. A polynucleotide encoding the polypeptide of any one of claims 1-32 or the nanostructure of any one of claims 33-36.

38. The polynucleotide of claim 37 is a messenger RNA (mRNA).

39. A lipid nanoparticle (LNP) comprising an mRNA encoding the polypeptide of any one of claims 1-32 or the nanostructure of any one of claims 33-36.

40. A pharmaceutical composition comprising the polypetide of any one of claims 1-32, the nanostructure of any one of claims 33-36, the polynucleotide of any one of claims 37-38 or the LNP of claim 39.

41. A method of generating an immune response to CMV gB protein in a subject in need thereof, comprising administering to the subject, an effective amount of the pharmaceutical composition of claim 40.

42. A method of treating or preventing CMV infection in a subject in need thereof, comprising administering to the subject, an effective amount of the pharmaceutical composition of claim 40.

43. An expression vector comprising the polynucleotide of any one of claims 37-38 operatively linked to a suitable control sequence.

44. A host cell comprising the polynucleotide of any one of claims 37-38, the expression vector of claim 43, and/or the polypeptide of any proceeding claim.

45. A polypeptide comprising an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO:3 residues 91-702, or an antigenic fragment thereof, wherein (a) residues 698-702 are optional if they do not include a mutation; (b) SEQ ID NO:3 residue 456 is absent, and (c) the polypeptide or antigenic fragment thereof comprises 1 or more combination of mutations relative to SEQ ID NO: 1 or SEQ ID NO:3 residues 91-702 listed in Table 2, Table 3, or Table 4.

46. The polypeptide of claim 45, wherein the polypeptide or antigenic fragment thereof comprises 1 or more combination of mutations relative to SEQ ID NO: 1 or SEQ ID NO: 3 residues 91-702 listed in Table 2 or Table 3.

47. The polypeptide of claim 45, wherein the polypeptide or antigenic fragment thereof comprises 1 or more combination of mutations relative to SEQ ID NO: 1 or SEQ ID NO: 3 residues 91-702 listed in Table 2.

48. The polypeptide of any one of claims 45-47, wherein the polypeptide or antigenic fragment thereof further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more mutations relative to relative to SEQ ID NO: 1 or SEQ ID NO: 3 residues 91-702 listed in Table 5.

49. The polypeptide of any one of claims 45-48, wherein the polypeptide or antigenic fragment thereof further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or all 10 mutations relative to relative to SEQ ID NO: 1 or SEQ ID NO:3 residues 91-702 listed in Table 6.

50. The polypeptide of any one of claims 45-48, wherein the polypeptide or antigenic fragment thereof further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or all 12, relative to relative to SEQ ID NO: 1 or SEQ ID NO:3 residues 91-702 listed in Table 7.

51. The polypeptide of any one of claims 45-50, wherein the polypeptide or antigenic fragment thereof further comprises a combination of mutations relative to relative to SEQ ID NO: 1 or SEQ ID NO:3 residues 91-702 listed in Table 8.

52. The polypeptide of any one of claims 45-51, comprising an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence selected from the sequences listed in Table 9, or an antigenic fragment thereof.

53. The polypeptide of any one of claims 45-52, comprising an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence selected from the sequences listed in Table 10, or an antigenic fragment thereof.

54. The polypeptide of any one of claims 45-53, further comprising any other functional domain as appropriate for an intended use, including but not limited to a secretion signal located at the N-terminus of the polypeptide, wherein the signal sequence may be any suitable signal sequence as appropriate for an intended use.

55. A fusion protein comprising: (a) the polypeptide or antigenic fragment thereof of any one of claims 45-54; and (b) a multimerization domain.

56. The fusion protein of claim 55, wherein the multimerization domain comprises a polypeptide with sequence identity of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% to the sequence of a protein listed in Table 12, wherein residues in parentheses are optional.

57. A composition, comprising one or more of the polypeptides or fusion proteins of any preceding claim linked to a scaffold.

58. The composition of claim 57, wherein the scaffold comprises a protein scaffold.

59. The composition of claim 58, wherein the polypeptide is covalently linked to a protein subunit of the protein scaffold to form a fusion protein.

60. The composition of claim 59, wherein the protein subunit of the protein scaffold comprises a polypeptide with sequence identity of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% to the sequence of a protein listed in Table 12, wherein residues in parentheses are optional.

61. A nucleic acid encoding the polypeptide or fusion protein of any preceding claim.

62. An expression vector comprising the nucleic acid of claim 61 operatively linked to a suitable control sequence.

63. A host cell comprising the nucleic acid of claim 61, the expression vector of claim 62, and/or the polypeptide or fusion protein of any preceding claim.

64. A pharmaceutical composition, comprising (a) one or more of the polypeptides, fusion protein, composition, nucleic acid, expression vector, and/or the host cell of any preceding claim; and (b) a pharmaceutically acceptable carrier.

65. A vaccine comprising (a) one or more of the polypeptides, fusion protein, composition, nucleic acid, expression vector, and/or the host cell of any preceding claim; and (b) a pharmaceutically acceptable carrier.

66. The vaccine of claim 65 further comprising any other component as appropriate for an intended use, including but not limited to any other CMV antigens, including but not limited to one or more of CMV proteins gH, gL, UL128, UL130 and UL131, or antigenic portions thereof; or a pentamer complex of CMV proteins gH, gL, UL128, UL130 and UL131 or antigenic portions thereof.

67. A method for treating or limiting development of an CMV infection, comprising administering to a subject in need thereof an amount effective to treat or limit development of the CMV infection of a polypeptide, fusion protein, composition, vaccine, nucleic acid, expression vector, host cell, pharmaceutical composition, and/or vaccine of any preceding claim.

68. A composition, method, or use as described herein.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0038] FIG. 1 shows structural representation of regions targeted in prefusion CMV gB structure for addition of engineered mutations. A) Prefusion structure of CMV gB (PDB 7KDP), with only the ectodomain shown. Highlighted regions contain sets of stabilizing mutations according to the names displayed. Without being bound by theory, mutations listed as Bg-KOs are used to reduce aggregation, improve expression and/or prevent cleavage by furin. B) Postfusion structure of CMV gB (PDB 7KDD).

[0039] FIG. 2 shows addition of glycan at N165. A) N165 (shown with spheres) is solvent-exposed in the prefusion structure (PDB 7KDP, shown with transparent surface). Two views are shown, one of which is further zoomed-in (right). B) N165 (shown with spheres) is buried in the postfusion structure. Two views are shown, one from the side of gB (left) and one from the bottom of gB looking down the 3-fold axis (right).

[0040] FIG. 3 shows recombinant gB designs including at least the soluble ectodomain of gB from the Towne strain from residues 20-698, with some designs containing additional residues up to residue 704. All designs tested featured an exogenous N-terminal signal peptide and a C-terminal octa-histidine tag, with some designs also featuring an exogenous trimerization domain between the C-terminus of the gB ectodomain and the octa-histidine tag.

[0041] FIG. 4 shows chromatographic elution profiles of multiple gB antigens featuring designed mutations, showing homogenous profiles consistent with expected elution profiles for trimeric gB antigens. A comparable gB antigen lacking designed mutations is shown as a reference.

[0042] FIG. 5 shows DLS of multiple gB antigens featuring designed mutations, showing distinctly different hydrodynamic diameters and polydispersities, which is indicative of conformational changes. A comparable gB antigen lacking designed mutations is shown as a reference.

[0043] FIG. 6 shows nanoDSF of multiple gB antigens featuring designed mutations, with nanoDSF measured using intrinsic tryptophan fluorescence. A comparable gB antigen lacking designed mutations is shown as a reference.

[0044] FIG. 7 shows NS-EM micrographs of multiple gB antigens featuring designed mutations, showing disruption of the postfusion state. A comparable gB antigen lacking designed mutations is shown as a reference, which more monodispersely features an elongated profile consistent with the postfusion state.

[0045] FIG. 8 shows NS-EM 2D class averages of one gB antigen featuring designed mutations and a comparable gB antigen lacking designed mutations.

DETAILED DESCRIPTION

Definitions

[0046] Throughout the disclosure, reference is made to particular features (including method steps). It is to be understood that the disclosure includes all possible combinations of such particular features. For example, where a particular feature is disclosed in the context of a particular aspect or embodiment, or a particular claim, that feature can also be used, to the extent possible, in combination with and/or in the context of other particular aspects and embodiments.

[0047] Where reference is made herein to a method comprising two or more defined steps, the defined steps can be carried out in any order or simultaneously (except where the context excludes that possibility), and the method can include one or more other steps which are carried out before any of the defined steps, between two of the defined steps, or after all the defined steps (except where the context excludes that possibility).

[0048] The practice of the technology will employ, unless indicated specifically to the contrary, conventional methods of chemistry, biochemistry, organic chemistry, molecular biology, microbiology, recombinant DNA techniques, genetics, immunology, and cell biology that are within the skill of the art, many of which are described below for the purpose of illustration. Such techniques are explained fully in the literature. It is to be understood that this disclosure is not limited to the particular methodology, protocols, and reagents described, as these may vary, depending upon the context in which they are used by those of skill in the art.

[0049] All publications and patents mentioned herein are hereby incorporated by reference in their entirety as if each individual publication or patent was specifically and individually indicated to be incorporated by reference. In case of conflict, the present application, including any definitions herein, controls. However, mention of any reference, article, publication, patent, patent publication, and patent application cited herein is not, and should not be taken as an acknowledgment, or any form of suggestion, that they constitute valid prior art or form part of the common general knowledge in any country in the world.

[0050] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. Specifically, features described in one section may be combined with features in any other section of the description.

[0051] While illustrative embodiments are described and depicted, it will be appreciated that various changes can be made to these illustrative embodiments without departing from the spirit and scope of the invention.

[0052] Unless defined otherwise herein, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Various scientific dictionaries that include the terms included herein are well known and available to those in the art. Although any methods and materials similar or equivalent to those described herein find use in the practice or testing of the disclosure, some preferred methods and materials are described. Accordingly, the terms defined immediately below are more fully described by reference to the specification as a whole.

[0053] The singular forms a, an, and the are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0054] As used herein, the term about means a range of values including the specified value, which a person of ordinary skill in the art would consider reasonably similar to the specified value. In some embodiment, about means within a standard deviation using measurements generally acceptable in the art. In some embodiment, about means a range extending to +/10%, +/5%, +/3%, or +/1% of the specified value.

[0055] The term at least followed by a number is used herein to denote the start of a range beginning with that number (which may be a range having an upper limit or no upper limit, depending on the variable being defined). For example, at least 1 means 1 or more than 1.

[0056] The term at most followed by a number is used herein to denote the end of a range ending with that number (which may be a range having 1 or 0 as its lower limit, or a range having no lower limit, depending upon the variable being defined). For example, at most 4 means 4 or less than 4, and at most 40% means 40% or less than 40%. When, in this specification, a range is given as (a first number) to (a second number) or (a first number)(a second number) this means a range whose lower limit is the first number and whose upper limit is the second number. For example, 25 to 100 mm means a range whose lower limit is 25 mm, and whose upper limit is 100 mm.

[0057] Throughout this specification, unless the context requires otherwise, the words comprise, comprises and comprising will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements. For example, a composition comprising (or which comprises) components A, B, and C can consist of (i.e., contain only) components A, B, and C, or can contain not only components A, B, and C but also one or more other components.

[0058] As used herein, the term consisting of refers to including, and limited to, whatever follows the phrase consisting of, Thus, the phrase consisting of indicates that the listed elements are required or mandatory, and that no other elements may be present. By consisting essentially of is meant including any elements listed after the phrase, and limited to other elements that do not interfere with or contribute to the activity or action specified in the disclosure for the listed elements. Thus, the phrase consisting essentially of indicates that the listed elements are required or mandatory, but that no other elements are optional and may or may not be present depending upon whether or not they affect the activity or action of the listed elements.

[0059] The term protein nanostructure, as used herein, refers to symmetric protein assemblies in which the subunits self-assemble in aqueous solution, without requiring lipids or macromolecules other than the protein nanostructure for assembly. Illustrative protein nanostructures are described in Hsia et al. Nature 35:136-9 (2016) and Bale et al. Science 353:389-394 (2016). In some embodiments, the protein nanostructure is a one-component protein nanostructure, in which a single polypeptide type provides the building blocks to self-assemble to form the protein nanostructure. In some embodiments, the protein nanostructure is a two-component protein nanostructure, in which two polypeptide types provide the building blocks to self-assemble to form the protein nanostructure. In some embodiments, the polypeptide types include an assembly domain, which causes the polypeptides to form symmetric dimeric, trimeric, tetrameric, hetaxameric components, or another multimeric component. In a two-component nanostructure, the two components differ in the selection of an assembly domain. In some embodiments, the assembly domain of the first polypeptide type causes the polypetide to form a trimer; and the assembly domain of the second polypeptide type causes the polypetide to form a pentamer.

[0060] In sone embodiments of, or relating to, one-component nanostructures, two or more copies of the component further symmetrically self-assemble to form the nanostructure. In some embodiments of, or relating to, two-component nanostructures, two or more of each of the two, different components symmetrically self-assemble to form the nanostructure.

[0061] The protein nanostructures disclosed herein can display potentially antigenic polypeptides intended to elicit immune responses to viruses and can be administered as vaccines. In some embodiments, the components include atrimer of antigenic polypeptides.

[0062] The vaccines of the present disclosure are useful for preventing and/or decreasing the severity of viral infection.

[0063] The term ferritin protein nanostructure, as vised herein, refers to using ferritin to generate a symmetric, protein-based protein nanostructure using naturally occurring ferritin sequences, or engineered variants thereof. Ferritin-based protein nanostructures are prepared by fusing an antigen to the ferritin molecule, Illustrative ferritin protein nanostructures are described (vising the term nanoparticle rather than protein nanostructure) in U.S. Pat. Appl. Pub. No. US 2018/0021258 A1 and US 2019/033027 A1, the contents of which are incorporated by reference herein. Further, the molecular architecture of ferritin, which consists of 24 subunits assembling into an octahedral cage with 432 symmetry has the potential to display multimeric antigens on its surface.

[0064] As used herein, the term assembly domain refers to the portion of a subunit of a component involved in forming the protein nanostructure through intra-component interations and interactions with either other copies of the same component (in a one-component nanostructure) or with other components (e.g., in a two-component nanostructure).

[0065] The term icosahedral particle refers to protein nanostructures having a core with icosahedral symmetry. 153 refers to an icosahedral particle constructed from pentamers and trimers. 152 refers to an icosahedral particle constructed from pentamers and dimers. T33 refers to a tetrahedral particle constructed from two sets of trimers. T32 refers to a tetrahedral particle constructed from trimers and dimers.

[0066] The potentially antigenic polypeptides may be attached to the core of the protein nanostructure either non-covalently or covalently, including as a fusion protein or by other means disclosed herein. Multimeric polypeptides may optionally be displayed along a symmetry axis of the protein nanostructure. Also provided are proteins and nucleic acid molecules encoding such proteins, formulations, and methods of use.

[0067] The term antigen or immunogen refers to its plain and ordinary meaning of a compound or composition that induces an immune response, cellular or humoral, e.g., cytotoxic T lymphocyte (CTL) response, a B cell response (for example, production of antibodies that specifically bind the epitope), an NK cell response or any combinations thereof, when administered to or expressed in an immunocompetent subject. Antigens can include polypeptides (including glycoproteins). In some embodiment, an antigen is a polypeptide or polypeptide complex that elicits an immune response. For example, an antigen can include one or more immunogenic epitopes associated with a viral pathogen. The tern antigen, as used herein, is not limited to the portion of the polypeptide or polypeptide complex that contains antigenic epitopes. An epitope or antigenic determinant refers to its plain an ordinary meaning as the part of an antigen that is recognized by the immune system, specifically by antibodies, B cells, or T cells.

[0068] The terms polypeptide, peptide and protein are used interchangeably herein to refer to a polymer of amino acid residues and optionally one or more post-translational modifications (e.g., glycosylation) and/or other modifications.

[0069] The term isolated when applied to a polynucleotide or polypeptide, denotes that the polynucleotide or polypeptide is essentially free of other cellular components with which it is associated in the natural state. It can be, for example, in a homogeneous state and may be in either a dry or aqueous solution. Purity and homogeneity are typically determined using analytical chemistry techniques such as polyacrylamide gel electrophoresis or high-performance liquid chromatography. A polynucleotide or polypeptide that is the predominant species present in a preparation is substantially purified.

[0070] The terms virus or virus particle are used according to its plain ordinary meaning within Virology and refers to a virion including the viral genome (e.g., DNA, RNA, single strand, double strand), viral capsid and associated proteins, and in the case of enveloped viruses (e.g., herpesvirus), an envelope including lipids and optionally components of host cell membranes, and/or viral proteins.

[0071] The term viral infection or viral disease refers to a disease or condition that is caused by a virus, such as by Cytomegalovirus (CMV).

[0072] The ten-n ectodomain refers to the portion of a transmembrane protein or glycoprotein that, in the native state of the protein, is on the outside of the cellular or viral membrane.

[0073] The term variant refers to a polypeptide or polynucleotide having one or more insertions, deletions, or amino acid substitutions relative to a reference polypeptide or polynucleotide.

[0074] The term antigenic variant refers to a variant that has one or more epitopes in common with a reference polypeptide and/or generates the same or similar immune response when administered to a subject as a reference polypeptide.

[0075] The term functional variant refers to a variant that exhibits at least some of the activity as a reference polypeptide. For example, a functional variant of an assembly domain is able to promote multimerization and self-assembly to the same extent, or to similar extent, as a reference assembly domain and/or is able to multimerize and assembly with the same cognate assembly domains as a reference assembly domain.

[0076] As used herein, the term bound when referring to two moieties means the two moieties are bonded, wherein the bond or bonds connecting the two moieties may be covalent or non-covalent. In some embodiment, the two moieties are covalently bonded to each other (e.g., directly or through a covalently bonded intermediary (a linker)). In some embodiment, the two moieties are non-covalently bonded (e.g., through ionic bond(s), Van der Waal's bond(s)/interactions, hydrogen bond(s), polar bond(s), or combinations or mixtures thereof). For a fusion polypeptide that is N-terminally and C-terminally joined the linker can be a peptide bond or peptide of any length.

[0077] The term domain refers to refers to any portion of a polypeptide that adopts a tertiary structure.

[0078] The terms multimerization domain and multimerize refer to the ability of a polypeptide, or domain of a polypeptide, to form a tertiary structure with another polypeptide of domain of a polypeptide. In some embodiments, multimerization domains can form dimers, trimers, tetramers, pentamers, or hexamers and/or to form heteromers with other multimerization domains.

[0079] The ten-n trimerization domain refers to a multimerization domain that forms trimers.

[0080] The term fragment refers to a polypeptide having one or more N-terminal or C-terminal truncations compared to a reference polypeptide.

[0081] The term functional fragment refers to a fragment that retains at least one function of its reference polypeptide.

[0082] The terms helix or helical refer to an -helical secondary structure in a polypeptide that is known to occur, or predicted to occur. For example, a sequence may be described as helical when computational modeling suggests the sequence is likely to adopt a helical conformation.

[0083] The term component refers to a protein, or protein complex, capable of assembly into a protein nanostructure under appropriate conditions (e.g., a fusion protein comprising an assembly domain).

[0084] The term vaccine refers to a composition that can provide active acquired immunity to and/or therapeutic effect (e.g., treatment) of a particular disease or a pathogen. A vaccine typically contains one or more agents that can induce an immune response in a subject against a pathogen or disease, i.e., a target pathogen or disease. The immunogenic agent stimulates the body's immune system to recognize the agent as a threat or indication of the presence of the target pathogen or disease, thereby inducing immunological memory so that the immune system can more easily recognize and destroy any of the pathogen on subsequent exposure. Vaccines can be prophylactic (e.g., preventing or ameliorating the effects of a future infection by any natural or pathogen, or of an anticipated occurrence of cancer in a predisposed subject) or therapeutic (e.g., treating; cancer in a subject who has been diagnosed with the cancer). The administration of vaccines is referred to vaccination.

[0085] The term pharmaceutically acceptable excipients and pharmaceutically acceptable carrier refer to a substance that aids the administration of an active agent to and absorption by a subject and can be included in the compositions of the present disclosure without causing a significant adverse toxicological effect on the patient and can mean excipients approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, lactated Ringer's, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethycellulose, polyvinyl pyrrolidine, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and/or aromatic substances and the like that do not deleteriously react with the compounds of the disclosure. One of skill in the art will recognize that other pharmaceutical excipients are useful in the present disclosure.

[0086] The term adjuvant refers to a pharmaceutically acceptable substance that enhances the immune response to an antigen when co-administered with the antigen or administered before, during, or after administration of the antigen to a subject. In some cases, the LNP polynucleotide (e.g., mRNA) can serve as the adjuvant.

[0087] The term TLR4 immunostimulant refers to an adjuvant that stimulates Toll-like Receptor 4 (TLR4) in the immune cells of a subject to modulate an immune response. Exemplary TLR4 immunostimulants include, but are not limited to, Monophosphoryl Lipid A (MPL), Glucopyranosyl Lipid A (GLA), and/or synthetic lipid A (SLA). In some embodiment, the antigen is a TLR4 immunostimulant.

[0088] The term effective amount refers to the amount of a composition that, when administered to a patient for treating a state, disorder or condition is sufficient to effect such treatment or when administered to a patient for generating an immune response is sufficient to generate such an immune response. The exact amounts will depend on the active ingredient, the state, disorder, or condition to be treated and its severity, and the age, weight, physical condition and responsiveness of the subject to be treated, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Liebennan, Pharmaceutical Dosage Forms (vols. 1-3, 1992); Lloyd, The Art, Science and Technology of Pharmaceutical Compounding (1999); Pickar, Dosage Calculations (1999); and Remington: The Science and Practice of Pharmacy, 20th Edition, 2003, Gennaro, Ed., Lippincott, Williams & Wilkins).

[0089] The terms immunization and immunizing refer to administering a composition to a subject in an amount sufficient to elicit, after one or more administering steps, a desired immune response, immunization may comprise between one and ten, or more administrations (e.g., injections) of the composition, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more administrations. The first administration may elicit no detectable immune response as generally each subsequent administration will boost the immune response generated by prior administrations. The term immunizing as used herein includes post-exposure prophylaxis.

[0090] The term protective immune response refers to an immune response that prevents and/or reduces the severity of infection with a pathogen when the subject is later challenged with the pathogen, or to an immune response that generates a level of immune response that correlates with protection. For example, vaccination may generate a protective immune response if it results in production, in the plasma or serum, of the subject (e.g., human, pet, or agricultural animal), of neutralizing antibodies that protect the subject against subsequent infection and/or are present in a quantity observed to confer protection upon test subjects (e.g., New Zealand White (NZW) rabbits).

[0091] The term polyclonal antibody response refers to an antibody response comprising antibodies having more than one specificities and/or variation in their antibody sequences.

[0092] The term neutralizing (e.g., neutralizing antibody response) refers to antibodies that prevent infection and/or reduce the level of infection by a pathogen. A neutralizing antibody response can be measured either in in vitro assays (e.g., infection of cells in culture by a pathogen in the presence of the antibody) or in an in vivo assay (e.g., by determining a protective dose of an antibody through administering the antibody to a subject prior to challenge with an infective dose of a pathogen).

[0093] An antibody binds to or is specific to or specifically binds (used interchangeably herein) to a target (e.g., viral protein) are terms well understood in the art, and methods to determine such specific or preferential binding are also well known in the art. A molecule is said to exhibit specific binding or preferential binding if it reacts or associates more frequently, more rapidly, with greater duration and/or with greater affinity with a particular cell or substance than it does with alternative cells or substances.

[0094] The term post-exposure prophylaxis refers to administering an antigen (e.g., a vaccine) to a subject previously exposed to and/or infected with a pathogen in order to elicit an immune response to protect against infection by the pathogen and/or decrease the severity of one or more symptoms of infection by the pathogen.

[0095] The term administering refers to providing a composition to a subject in a manner that permits the composition to have its intended effect. Administration for vaccination or post-exposure prophylaxis may be performed by intramuscular injection, intravenous injection, intraperitoneal injection, or any other suitable route.

[0096] Co-administer means that a composition described herein is administered at the same time, just prior to, or just after the administration of one or more additional therapies. The composition provided herein can be administered alone or can be coadministered to the subject. Co-administration is meant to include simultaneous or sequential administration of the compounds individually or in combination (more than one compound). Thus, the preparations can also be combined, when desired, with other active substances (e.g., to reduce metabolic degradation).

[0097] The term subject refers to a human or non-human animal to which a composition may be administered for vaccination, treatment, or other purpose. In some embodiment, the non-human animal is a non-human primate, rabbit, hamster, gerbil, pig, cow, sheep, goat, guinea pig, rat, mouse, squirrel, wolf, fox, horse, zebra, giraffe, elephant, cat, dog, llama, or ferret.

[0098] The term manufacturing refers to production of a recombinant polynucleotide, polypeptide, delivery vehicle, or protein nanostructure at any scale, including at least 25-mL, 50-mL, 1-L, 1,000-L, 50,000-L, or greater scale.

[0099] The terms culturing and culture medium refers to standard cell culture and recombinant protein expression techniques.

[0100] The term host cell refers to any cell capable of use in expression of a recombinant polypeptide or polynucleotide.

[0101] The term secretes refers to the ability of host cells to release expressed polypeptides into the media in which they are cultured.

[0102] The term signal sequence refers to a polypeptide sequence, typically at the N terminus of a polypeptide, expressed in a host cell that directs the polypeptide to a particular cellular compartment. A signal sequence may be a secretion signal to cause the host cell to secrete the polypeptide into the media in which with host cell is cultured. The signal sequence can be the native signal sequence, a signal sequence that occurs in nature as part of the polypeptide. The signal sequence can be a sequence that does not naturally occur with the polypeptide as found in nature. Various signal sequences are known and it is within the skill of an ordinary artisan to select an appropriate signal sequence.

[0103] The term purify refers to separating a molecule from other substances present in a composition. Polypeptides may be purified by affinity (e.g., to an antibody or to a tag, e.g., using a His-tag capture resin), by charge (e.g., ion-exchange chromatography), by size (e.g., preparative ultracentrifugation, size exclusion chromatography), or otherwise.

[0104] The terms polynucleotide and nucleic acid, used interchangeably herein, refer to a polymeric form of nucleotides of more than about 100 nucleotides, either ribonucleotides or deoxyribonucleotides. Thus, this term includes, but is not limited to, single-, double-, or multi-stranded DNA or RNA, mRNA, genomic DNA, cDNA, DNA-RNA hybrids, or a polymer comprising purine and pyrimidine bases or other natural, chemically or biochemically modified, non-natural, or derivatized nucleotide bases. Oligonucleotide generally refers to polynucleotides of between about 5 and about 100 nucleotides of single- or double-stranded DNA. However, for the purposes of this disclosure, there is no upper limit to the length of an oligonucleotide. Oligonucleotides are also known as oligoners or oligos and may be isolated from genes, or chemically synthesized by methods known in the art. The terms polynucleotide and nucleic acid should be understood to include, as applicable to the embodiments being described, single-stranded (such as sense or antisense) and double-stranded polynucleotides.

[0105] The terms identity, identical, and sequence identity in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino acid residues or nucleotides that are the same (i.e., about 60% identity, preferably 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or higher identity over a specified region). Such sequences are then said to be substantially identical. This definition also refers to, or may be applied to, the compliment of a test sequence. The definition also includes sequences that have deletions and/or additions, as well as those that have substitutions. The term amino acid substitution refers to replacing a single amino acid in a sequence with another amino acid residue. The standard form of abbreviations for amino acid substitution are used. For example, V94R refers to substitution of valine (V) in a reference sequence with arginine (R). The abbreviation Arg94 refers to any sequence in which the 94th residue, relative to a reference sequence, is arginine (Arg). As described below, the preferred algorithms can account for gaps and the like. Preferably, identity exists over a region that is at least about 25 amino acids or nucleotides in length, or more preferably over a region that is 50-100 amino acids or nucleotides in length. For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. The term reference sequence refers to a molecule to which a test sequence is compared.

[0106] Methods of sequence alignment for comparison and determination of percent sequence identity is well known in the art. Optimal alignment of sequences for comparison can be conducted, e.g., by the homology alignment algorithm of Needleman and Wunsch, J. Mol. Biol. 48:443-53 (1970).

[0107] An amino acid or nucleotide base position is denoted by a number that sequentially identifies each amino acid (or nucleotide base) in the reference sequence based on its position relative to the N-terminus (or 5-end). Due to deletions, insertions, truncations, fusions, and the like that must be taken into account when determining an optimal alignment, in general the amino acid residue number in a test sequence determined by simply counting from the N-terminus will not necessarily be the same as the number of its corresponding position in the reference sequence. For example, in a case where a variant has a deletion relative to an aligned reference sequence, there will be no amino acid in the variant that corresponds to a position in the reference sequence at the site of deletion. Where there is an insertion in an aligned reference sequence, that insertion will not correspond to a numbered amino acid position in the reference sequence. In the case of truncations or fusions there can be stretches of amino acids in either the reference or aligned sequence that do not correspond to any amino acid in the corresponding sequence.

[0108] The terms numbered with reference to or corresponding to, when used in the context of the numbering of a given amino acid or polynucleotide sequence, refers to the numbering of the residues of a specified reference sequence when the given amino acid or polynucleotide sequence is compared to the reference sequence.

[0109] The term treating means one or more of relieving, alleviating, delaying, reducing, reversing, improving, or managing at least one symptom of a condition in a subject. The term treating may also mean one or more of arresting, delaying the onset (i.e., the period prior to clinical manifestation of the condition) or reducing the risk of developing or worsening a condition.

EMBODIMENTS

[0110] Provided are polynucleotides that encode a polypeptide monomer of a trimeric component for a one-component protein nanostructure. Such protein nanostructures, when assembled, display viral protein trimers as antigens.

gB Protein

[0111] In some embodiment, the protein nanostructure comprises a Cytomegalovirus gB protein monomer or variant thereof. Illustrative gB proteins are provided in Table 1. Signal peptides are in parentheses and N-terminal residues, membrane-proximal region, transmembrane, and cytoplasmic residues are underlined. In each case, the underlined portions may be truncated to leave an ectodomain; and the same or different signal peptide may be used. In some embodiment, the Cytomegalovirus gB viral protein monomer comprises a polypeptide sequence at least 70%, at least 80%, at least 90%, at least 95%, at least 98% identical to any one of the polypeptide sequences in Table 1, or to the ectodomain thereof. In some embodiments, the Cytomegalovirus gB viral protein monomer comprises a polypeptide sequence at least 70%, at least 80%, at least 90%, at least 95%, at least 98%, or at least 99% identical to any one of the polypeptide sequences disclosed therein.

TABLE-US-00001 TABLE1 HCMVGBPROTEIN SEQ ID NO: Sequence Virus 7 (MESRIWCLVVCVNLCIVCL)GAAVSSSSTRGTSATHSHHSS hCMVgB, HTTSAAHSRSGSVSQRVISSQTVSHGVNETIYNTTLKYGDVV Townestrain,full GVNTTKYPYRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEG length IMVVYKRNIVAHTFKVRVYQKVLTFRRSYAYIHTTYLLGSNT EYVAPPMWEIHHINSHSQCYSSYSRVIAGTVEVAYHRDSYEN KTMQLMPDDYSNTHSTRYVTVKDQWHSRGSTWLYRETCNINC MVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRNASYEGE NADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISW DIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKMTATEL SKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNV SVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTKR STDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQI AEAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGD VLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENEAN SSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEY VDYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQK ELRSSNVEDLEEIMREFNSYKQRVKYVEDKVVDPLPPYLKGL DDLMSGLGAAGKAVGVAIGAVGGAVASVVEGVATFLKNPFGA FTIILVAIAVVIIIYLIYTRQRRLCMQPLQNLFPYLVSADGT TVTSGNTKDTSLQAPPSYEESVYNSGRKGPGPPSSDASTAAP PYTNEQAYQMLLALVRLDAEQRAQQNGTDSLDGQTGTQDKGQ KPNLLDRLRHRKNGYRHLKDSDEEENV 2 (MESRIWCLVVCVNICIVCL)GAAVSSSSTRGTSATHSHHSS hCMVgB, HTTSAAHSRSGSVSQRVTSSQTVSHGVNETIYNTTLKYGDVV Merlinstrain,full GVNTTKYPYRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEG length IMVVYKRNIVAHTFKVRVYQKVLTFRRSYAYIHTTYLLGSNT EYVAPPMWEIHHINSHSQCYSSYSRVIAGTVEVAYHRDSYEN KTMQLMPDDYSNTHSTRYVTVKDQWHSRGSTWLYRETCNINC MVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRNASYFGE NADKFFIFPNYTIVSDEGRPNSALETHRIVAFLERADSVISW DIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFL SKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNV SVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTKR STDGNNATHISNMESVHNLVYAQLQFTYDTLRGYINRALAQI AEAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGD VLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFAN SSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEY VDYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQK ELRSSNVEDLEEIMREENSYKQRVKYVEDKVVDPLPPYLKGL DDLMSGLGAAGKAVGVAIGAVGGAVASVVEGVATFLKNPFGA FTIILVAIAVVIITYLIYTRQRRLCTQPLQNLFPYLVSADGT TVTSGSTKDTSLQAPPSYEESVYNSGRKGPGPPSSDASTAAP PYTNEQAYQMLLALARLDAEQRAQQNGTDSLDGRTGTQDKGQ KPNLLDRLRHRKNGYRHLKDSDEEENV 3 (MESRIWCLVVCVNICIVCL)GAAVSSSSTSHATSSTHNGSH hCMVgB,strain TSRTTSAQTRSVYSQHVTSSEAVSHRANETIYNTTLKYGDVV AD169,full GVNTTKYPYRVCSMAQGTDLIRFERNIICTSMKPINEDLDEG length IMVVYKRNIVAHTFKVRVYQKVLTFRRSYAYIYTTYLLGSNT EYVAPPMWEIHHINKFAQCYSSYSRVIGGTVEVAYHRDSYEN KIMQLIPDDYSNTHSTRYVTVKDQWHSRGSTWLYRETCNINC MLTITTARSKYPYHFFATSTGDVVYISPFYNGTNRNASYFGE NADKFFIFPNYTIVSDFGRPNAAPETHRLVAFLERADSVISW DIQDEKNVICQLTFWEASERTIRSEAEDSYHESSAKMTATEL SKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNV SVFETSGGLVVFWQGIKQKSLVELERLANRSSINITHRTRRS TSDNNTTHLSSMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDV LGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANS SYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKE LRSSNVEDLEEIMREFNSYKQRVKYVEDKVVDPLPPYLKGLD DLMSGLGAAGKAVGVAIGAVGGAVASVVEGVATELKNPFGAF TIILVAIAVVIITYLIYTRQRRLCTQPLQNLFPYLVSADGTT VTSGSTKDTSLQAPPSYEESVYNSGRKGPGPPSSDASTAAPP YTNEQAYQMLLALARLDAEQRAQQNGTDSLDGQTGTQDKGQK PNLLDRLRHRKNGYRHLKDSDEEENV 4 GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQ hCMVgB TVSHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLI extracellular RFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQK domain,Towneor VLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYS Merlinstrain SYSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTV KDQWHSRGSTWLYRETCNLNCMVTITTARSKYPYHFFATSTG DVVDISPFYNGTNRNASYFGENADKFFIFPNYTIVSDFGRPN SALETHRLVAFLERADSVISWDIQDEKNVTCQLTFWEASERT IRSEAEDSYHESSAKMTATFLSKKQEVNMSDSALDCVRDEAI NKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQGIKQKSL VELERLANRSSLNLTHNRTKRSTDGNNATHISNMESVHNLVY AQLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVEKELSKI NPSAILSAIYNKPIAARFMGDVLGLASCVTINQTSVKVLRDM NVKESPGRCYSRPVVIENFANSSYVQYGQLGEDNEILLGNHR TEECQLPSLKIFIAGNSAYEYVDYLFKRMIDLSSISTVDSMI ALDIDPLENTDERVLELYSQKELRSSNVEDLEEIMREFNSYK QRVKYVEDKV 5 GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQ hCMVgB TVSHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLI extracellular RFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQK domain,Towneor VLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYS Merlinstrain-C- SYSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTV terminusshorten KDQWHSRGSTWLYRETCNLNCMVTITTARSKYPYHFFATSTG DVVDISPEYNGTNRNASYFGENADKFFIEPNYTIVSDEGRPN SALETHRLVAFLERADSVISWDIQDEKNVTCQLTFWEASERT IRSEAEDSYHFSSAKMTATFLSKKQEVNMSDSALDCVRDEAI NKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQGIKQKSL VELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVY AQLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKI NPSAILSAIYNKPIAAREMGDVLGLASCVTINQTSVKVLRDM NVKESPGRCYSRPVVIENFANSSYVQYGQLGEDNEILLGNHR TEECQLPSLKIFIAGNSAYEYVDYLFKRMIDLSSISTVDSMI ALDIDPLENTDERVLELYSQKELRSSNVE 6 GAAVSSSSTSHATSSTHNGSHTSRTTSAQTRSVYSQHVTSSE hCMVgB,strain AVSHRANETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLI AD169, RFERNIICTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQK extracellular VLTFRRSYAYIYTTYLLGSNTEYVAPPMWEIHHINKFAQCYS domain SYSRVIGGTVFVAYHRDSYENKTMQLIPDDYSNTHSTRYVTV KDQWHSRGSTWLYRETCNUNCMLTITTARSKYPYHFFATSTG DVVYISPFYNGTNRNASYFGENADKFFIFPNYTIVSDEGRPN AAPETHRLVAFLERADSVISWDIQDEKNVTCQLTFWEASERT IRSEAEDSYHESSAKMTATFLSKKQEVNMSDSALDCVRDEAI NKLQQIFNTSYNQTYEKYGNVSVFETSGGLVVEWQGIKQKSL VELERLANRSSLNITHRTRRSTSDNNTTHLSSMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVEKELSKIN PSAILSAIYNKPIAAREMGDVLGLASCVTINQTSVKVLRDMN VKESPGRCYSRPVVIFNFANSSYVQYGQLGEDNEILLGNHRT EECQLPSLKIFIAGNSAYEYVDYLEKRMIDLSSISTVDSMIA LDIDPLENTDERVLELYSQKELRSSNVEDLEEIMREFNSYKQ RVKYVEDKV 7 GAAVSSSSTSHATSSTHNGSHTSRTTSAQTRSVYSQHVTSSE hCMVgB,strain AVSHRANETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLI AD169, RFERNIICTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQK extracellular VLTERRSYAYIYTTYLLGSNTEYVAPPMWEIHHINKFAQCYS domain-C- SYSRVIGGTVFVAYHRDSYENKTMQLIPDDYSNTHSTRYVTV terminusshorten KDQWHSRGSTWLYRETCNLNCMLTITTARSKYPYHFFATSTG DVVYISPFYNGINRNASYFGENADKFFIEPNYTIVSDEGRPN AAPETHRLVAFLERADSVISWDIQDEKNVTCQLTFWEASERT IRSEAEDSYHFSSAKMTATFLSKKQEVNMSDSALDCVRDEAI NKLQQIFNTSYNQTYEKYGNVSVFETSGGLVVFWQGIKQKSL VELERLANRSSLNITHRTRRSTSDNNTTHLSSMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVEKELSKIN PSAILSAIYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMN VKESPGRCYSRPVVIENFANSSYVQYGQLGEDNEILLGNHRT EECQLPSLKIFIAGNSAYEYVDYLFKRMIDLSSISTVDSMIA LDIDPLENTDERVLELYSQKELRSSNVEDLEEIMREENSYKQ RVKYVEDKV

[0112] SEQ ID NO: 1 is the amino acid sequence for the cytomegalovirus (CMV) Towne strain glycoprotein B (gB). SEQ ID NO: 2 is the amino acid sequence for the CMV Merlin strain gB. SEQ ID NO 3 is the amino acid sequence for CMV strain AD169 gB, which shares about 95% identity to SEQ ID NO: 1 and has a deletion relative to SEQ ID NO: 1 and 2 at residue 456.

Amino Acid Substitutions

[0113] Provided are sequences of recombinant CMV gB proteins in which, while not wishing to be held by theory, amino acid substitutions have been added to disrupt the postfusion state and/or stabilize the prefusion state.

[0114] In some embodiment, the Cytomegalovirus gB protein includes one of the following combinations of mutations:

TABLE-US-00002 TABLE 2 Category 1 367I and 374F 686L and 690F 686L and 693V 690F and 693V 686L, 690F, and 693V 686L, 690F, 693V, and 694L 686L, 690F, 693V, and 694L, 700A 686L, 690F, 693V, 694L, 700A, and 702Q 218F, 591F, and 668A 218F, 591Y, and 668A 220S and 591F 220S and 591Y 552L and 591F 552L and 591Y 220S and 552L 220S and 552L, 591F 220S, 552L, 591Y 218F, 220S, 552L, 591F, and 668A 218F, 220S, 552L, 591Y, and 668A 217C and 587C 217C and 589C

TABLE-US-00003 TABLE 3 Category 2 217C, 587C, 686L, 690F, 693V 217C, 587C, 686L, 690F, 693V, 694L 217C, 587C, 686L, 690F, 693V, 694L, 700A 217C, 589C, 686L, 690F, 693V 217C, 589C, 686L, 690F, 693V, 694L 217C, 589C, 686L, 690F, 693V, 694L, 700A 217C, 587C, 591Y, 686L, 690F, 693V 217C, 587C, 591Y, 686L, 690F, 693V, 694L 217C, 587C, 591Y, 686L, 690F, 693V, 694L, 700A 217C, 589C, 591Y, 686L, 690F, 693V 217C, 589C, 591Y, 686L, 690F, 693V, 694L 217C, 589C, 591Y, 686L, 690F, 693V, 694L, 700A 217C, 587C, 591F, 686L, 690F, 693V 217C, 587C, 591F, 686L, 690F, 693V, 694L 217C, 587C, 591F, 686L, 690F, 693V, 694L, 700A 217C, 589C, 591F, 686L, 690F, 693V 217C, 589C, 591F, 686L, 690F, 693V, 694L 217C, 589C, 591F, 686L, 690F, 693V, 694L, 700A 220S, 552L, 686L, 690F, 693V 220S, 552L, 686L, 690F, 693V, 694L 220S, 552L, 686L, 690F, 693V, 694L, 700A 218F, 220S, 552L, 591F, 668A, 686L, 690F, 693V 218F, 220S, 552L, 591F, 668A, 686L, 690F, 693V, 694L 218F, 220S, 552L, 591F, 668A, 686L, 690F, 693V, 694L, 700A 218F, 220S, 552L, 591Y, 668A, 686L, 690F, 693V 218F, 220S, 552L, 591Y, 668A, 686L, 690F, 693V, 694L 218F, 220S, 552L, 591Y, 668A, 686L, 690F, 693V, 694L, 700A 218F, 591F, 668A, 686L, 690F, 693V 218F, 591F, 668A, 686L, 690F, 693V, 694L 218F, 591F, 668A, 686L, 690F, 693V, 694L, 700A 218F, 591Y, 668A, 686L, 690F, 693V 218F, 591Y, 668A, 686L, 690F, 693V, 694L 218F, 591Y, 668A, 686L, 690F, 693V, 694L, 700A

TABLE-US-00004 TABLE 4 Category 3 217C, 587C, 686L, 690F 217C, 589C, 686L, 690F 217C, 587C, 591Y, 686L, 690F 217C, 589C, 591Y, 686L, 690F 217C, 587C, 591F, 686L, 690F 217C, 589C, 591F, 686L, 690F 217C, 220S, 587C, 591F, 686L, 690F 217C, 220S, 587C, 591F, 686L, 690F, 693V 217C, 220S, 587C, 591F, 686L, 690F, 693V, 694L 217C, 220S, 587C, 591F, 686L, 690F, 693V, 694L, 700A 217C, 220S, 589C, 591F, 686L, 690F 217C, 220S, 589C, 591F, 686L, 690F, 693V 217C, 220S, 589C, 591F, 686L, 690F, 693V, 694L 217C, 220S, 589C, 591F, 686L, 690F, 693V, 694L, 700A 217C, 220S, 587C, 591Y, 686L, 690F 217C, 220S, 587C, 591Y, 686L, 690F, 693V 217C, 220S, 587C, 591Y, 686L, 690F, 693V, 694L 217C, 220S, 587C, 591Y, 686L, 690F, 693V, 694L, 700A 217C, 220S, 589C, 591Y, 686L, 690F 217C, 220S, 589C, 591Y, 686L, 690F, 693V 217C, 220S, 589C, 591Y, 686L, 690F, 693V, 694L 217C, 220S, 589C, 591Y, 686L, 690F, 693V, 694L, 700A 217C, 220S, 552L, 587C, 686L, 690F 217C, 220S, 552L, 587C, 686L, 690F, 693V 217C, 220S, 552L, 587C, 686L, 690F, 693V, 694L 217C, 220S, 552L, 587C, 686L, 690F, 693V, 694L, 700A 217C, 220S, 552L, 589C, 686L, 690F 217C, 220S, 552L, 589C, 686L, 690F, 693V 217C, 220S, 552L, 589C, 686L, 690F, 693V, 694L 217C, 220S, 552L, 589C, 686L, 690F, 693V, 694L, 700A 217C, 220S, 552L, 587C, 591F, 686L, 690F 217C, 220S, 552L, 587C, 591F, 686L, 690F, 693V 217C, 220S, 552L, 587C, 591F, 686L, 690F, 693V, 694L 217C, 220S, 552L, 587C, 591F, 686L, 690F, 693V, 694L, 700A 217C, 220S, 552L, 589C, 591F, 686L, 690F 217C, 220S, 552L, 589C, 591F, 686L, 690F, 693V 217C, 220S, 552L, 589C, 591F, 686L, 690F, 693V, 694L 217C, 220S, 552L, 589C, 591F, 686L, 690F, 693V, 694L, 700A 217C, 220S, 552L, 587C, 591Y, 686L, 690F 217C, 220S, 552L, 587C, 591Y, 686L, 690F, 693V 217C, 220S, 552L, 587C, 591Y, 686L, 690F, 693V, 694L 217C, 220S, 552L, 587C, 591Y, 686L, 690F, 693V, 694L, 700A 217C, 220S, 552L, 589C, 591Y, 686L, 690F 217C, 220S, 552L, 589C, 591Y, 686L, 690F, 693V 217C, 220S, 552L, 589C, 591Y, 686L, 690F, 693V, 694L 217C, 220S, 552L, 589C, 591Y, 686L, 690F, 693V, 694L, 700A 217C, 218F, 220S, 552L, 587C, 591F, 668A, 686L, 690F 217C, 218F, 220S, 552L, 587C, 591F, 668A, 686L, 690F, 693V 217C, 218F, 220S, 552L, 587C, 591F, 668A, 686L, 690F, 693V, 694L 217C, 218F, 220S, 552L, 587C, 591F, 668A, 686L, 690F, 693V, 694L, 700A 217C, 218F, 220S, 552L, 589C, 591F, 668A, 686L, 690F 217C, 218F, 220S, 552L, 589C, 591F, 668A, 686L, 690F, 693V 217C, 218F, 220S, 552L, 589C, 591F, 668A, 686L, 690F, 693V, 694L 217C, 218F, 220S, 552L, 589C, 591F, 668A, 686L, 690F, 693V, 694L, 700A 217C, 218F, 220S, 552L, 587C, 591Y, 668A, 686L, 690F 217C, 218F, 220S, 552L, 587C, 591Y, 668A, 686L, 690F, 693V 217C, 218F, 220S, 552L, 587C, 591Y, 668A, 686L, 690F, 693V, 694L 217C, 218F, 220S, 552L, 587C, 591Y, 668A, 686L, 690F, 693V, 694L, 700A 217C, 218F, 220S, 552L, 589C, 591Y, 668A, 686L, 690F 217C, 218F, 220S, 552L, 589C, 591Y, 668A, 686L, 690F, 693V 217C, 218F, 220S, 552L, 589C, 591Y, 668A, 686L, 690F, 693V, 694L 217C, 218F, 220S, 552L, 589C, 591Y, 668A, 686L, 690F, 693V, 694L, 700A 217C, 218F, 587C, 591F, 668A, 686L, 690F 217C, 218F, 587C, 591F, 668A, 686L, 690F, 693V 217C, 218F, 587C, 591F, 668A, 686L, 690F, 693V, 694L 217C, 218F, 587C, 591F, 668A, 686L, 690F, 693V, 694L, 700A 217C, 218F, 589C, 591F, 668A, 686L, 690F 217C, 218F, 589C, 591F, 668A, 686L, 690F, 693V 217C, 218F, 589C, 591F, 668A, 686L, 690F, 693V, 694L 217C, 218F, 589C, 591F, 668A, 686L, 690F, 693V, 694L, 700A 217C, 218F, 587C, 591Y, 668A, 686L, 690F 217C, 218F, 587C, 591Y, 668A, 686L, 690F, 693V 217C, 218F, 587C, 591Y, 668A, 686L, 690F, 693V, 694L 217C, 218F, 587C, 591Y, 668A, 686L, 690F, 693V, 694L, 700A 217C, 218F, 589C, 591Y, 668A, 686L, 690F 217C, 218F, 589C, 591Y, 668A, 686L, 690F, 693V 217C, 218F, 589C, 591Y, 668A, 686L, 690F, 693V, 694L 217C, 218F, 589C, 591Y, 668A, 686L, 690F, 693V, 694L, 700A 217C, 587C, 591Y 217C, 589C, 591Y 217C, 587C, 591F 217C, 589C, 591F 217C, 220S, 587C, 591F 217C, 220S, 589C, 591F 217C, 220S, 587C, 591Y 217C, 220S, 589C, 591Y 217C, 220S, 552L, 587C 217C, 220S, 552L, 589C 217C, 220S, 552L, 587C, 591F 217C, 220S, 552L, 589C, 591F 217C, 220S, 552L, 587C, 591Y 217C, 220S, 552L, 589C, 591Y 217C, 218F, 220S, 552L, 587C, 591F, 668A 217C, 218F, 220S, 552L, 589C, 591F, 668A 217C, 218F, 220S, 552L, 587C, 591Y, 668A 217C, 218F, 220S, 552L, 589C, 591Y, 668A 217C, 218F, 587C, 591F, 668A 217C, 218F, 589C, 591F, 668A 217C, 218F, 587C, 591Y, 668A 217C, 218F, 589C, 591Y, 668A 591Y, 686L, 690F 591Y, 686L, 690F, 693V 591Y, 686L, 690F, 693V, 694L 591Y, 686L, 690F, 693V, 694L, 700A 591F, 686L, 690F 591F, 686L, 690F, 693V 591F, 686L, 690F, 693V, 694L 591F, 686L, 690F, 693V, 694L, 700A 220S, 591F, 686L, 690F 220S, 591F, 686L, 690F, 693V 220S, 591F, 686L, 690F, 693V, 694L 220S, 591F, 686L, 690F, 693V, 694L, 700A 220S, 591Y, 686L, 690F 220S, 591Y, 686L, 690F, 693V 220S, 591Y, 686L, 690F, 693V, 694L 220S, 591Y, 686L, 690F, 693V, 694L, 700A 220S, 552L. 686L. 690F 220S, 552L, 591F, 686L, 690F 220S, 552L, 591F, 686L, 690F, 693V 220S, 552L, 591F, 686L, 690F, 693V, 694L 220S, 552L, 591F, 686L, 690F, 693V, 694L, 700A 220S, 552L, 591Y, 686L, 690F 220S, 552L, 591Y, 686L, 690F, 693V 220S, 552L, 591Y, 686L, 690F, 693V, 694L 220S, 552L, 591Y, 686L, 690F, 693V, 694L, 700A 218F, 220S, 552L, 591F, 668A, 686L, 690F 218F, 220S, 552L, 591Y, 668A, 686L, 690F 218F, 591F, 668A, 686L, 690F 218F, 591Y, 668A, 686L, 690F

[0115] Without adherence to theory, the above mutation combinations serve to disrupt the postfusion state and/or stabilize the prefusion state, thus making the polypeptides particularly useful as immunogens to induce an immune response against CMV, and/or for vaccinating against CMV. In some embodiments, residues 698-702 are optional if none of the mutations in the combination of mutations includes a mutation at residues 698-702.

[0116] Tables 2-4 list useful stabilizing gB mutations.

[0117] Table 5 lists additional mutations that are useful in combination with the mutations listed in Tables 2-4.

TABLE-US-00005 TABLE 5 Category 4 93I 93L 93M 96T 96V 96I 96L 221P 552M 552F 552Y 582P 584L 584M 584Y 603P 664F 664Y 664W 664S 664A 677T 678N 679N 679H 680E 681N 682S 684S 685Q 687A 688E 692S 695D 696R 699K

[0118] Table 6 lists additional mutations useful in combination with the mutations listed in Tables 2-5.

TABLE-US-00006 TABLE 6 Category 5 155G 156H 157R 206H 240A 240N 241T 242T 242H 246S 457S 460S

[0119] Table 7 lists additional mutations that are useful in combination with the mutations listed in Tables 2-6.

TABLE-US-00007 TABLE 7 Category 6 155H 155D 155E 155S 155N 156T 156D 156E 156Q 156K 156R 157D 157E 157S 157K 157N 157T 206G 206A 206S 206N 206Q 206K 206R 240S 240Q 240D 240E 240K 240R 240H 241S 241A 241G 241N 242S 242A 242G 242N 242Q 246G 246A 246N 246T 246Q 246K 246R 149T, 246T 149T, 246V 149T, 246N 149T, 246Q 149T, 246K 149T, 246R 149V, 246T 149V, 246V 149V, 246N 149V, 246Q 149V, 246K 149V, 246R 149I, 246T 149I, 246V 149I, 246N 149I, 246Q 149I, 246K 149I, 246R 457N 457Q 457A 457T 457G 457D 457E 459N 459Q 459A 459T 459G 459D 459E 460N 460Q 460A 460T 460G 460D 460E

[0120] Mutations listed in Table 8 are known to known volatile elements in the gB protein which make the protein difficult to express/produce/isolate.

TABLE-US-00008 TABLE 8 Category 7 156S, 157K, 240D, 242S, 246T 156D, 157K, 240S, 242S, 246T 156S, 157E, 240D, 242S, 246T 156D, 157E, 240S, 242S, 246T 156S, 157K, 240D, 242N, 246T 156D, 157K, 240S, 242N, 246T 156S, 157E, 240D, 242N, 246T 156D, 157E, 240S, 242N, 246T 156S, 157K, 240D, 242S, 246A 156D, 157K, 240S, 242S, 246A 156S, 157E, 240D, 242S, 246A 156D, 157E, 240S, 242S, 246A 156S, 157K, 240D, 242N, 246A 156D, 157K, 240S, 242N, 246A 156S, 157E, 240D, 242N, 246A 156D, 157E, 240S, 242N, 246A 149T, 156S, 157K, 240D, 242S, 246T 149T, 156D, 157K, 240S, 242S, 246T 149T, 156S, 157E, 240D, 242S, 246T 149T, 156D, 157E, 240S, 242S, 246T 149T, 156S, 157K, 240D, 242N, 246T 149T, 156D, 157K, 240S, 242N, 246T 149T, 156S, 157E, 240D, 242N, 246T 149T, 156D, 157E, 240S, 242N, 246T 149T, 156S, 157K, 240D, 242S, 246N 149T, 156D, 157K, 240S, 242S, 246N 149T, 156S, 157E, 240D, 242S, 246N 149T, 156D, 157E, 240S, 242S, 246N 149T, 156S, 157K, 240D, 242N, 246N 149T, 156D, 157K, 240S, 242N, 246N 149T, 156S, 157E, 240D, 242N, 246N 149T, 156D, 157E, 240S, 242N, 246N

[0121] In one aspect, the disclosure provides a polypeptide comprising an ectodomain of CMV gB in the prefusion conformation, wherein the ectodomain comprises, 1, 2, 4 or more amino acid substitutions selected from the group consisting of E686L, R693V, V694L, Y690F, K700A, V702E, D6791H, E681N, K695D, L680E, R685Q, V701L, E682S, F678N, N688E, V677T, D699K, F687A, M684S, Q692S, and Y696R. or substitutions at the same amino acid positions; the amino acid substitution S3671L the amino acid substitution T374F, or tie amino acid substitutions S367I and T374F, or substitutions at positions 367 and/or 374; 1, 2, 3, 4 or more amino acid substitutions selected from the group consisting of Q591F, Q591Y, S668A, Y218F, N220S, V552L, or substitutions at the same amino acid positions; the amino acid substitution E167T; and/or 1, 2, 3, 4 or more amino acid substitutions selected from the group consisting of C246S, 11157R, 1561-1, R457S, R460S, W240N, and Y242T at positions relative to SEQ ID NO: 1.

[0122] In some embodiments, the ectodomain comprises the amino acid subsitutions D217C and S587C. In some embodiments, the ectodomain comprises the amino acid subsitutions D217C and Y589C. 1012211n some embodiments, the ectodomain comprises 1, 2, 3, 4 or more amino acid substitutions selected from the group consisting of E686L, R693V, V694L, Y690F, K700A, V702E, D679H, E681N, K695D, L680E, R685Q. V701K, E682S, F678N, N688E. V677T. D699K, F687A, M684S, Q692S, and Y696R.

[0123] In some embodiments, the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, and Y690F. In some embodiments, the ectodomain comprises the amino acid substitutions E686L, R693, V694L, Y690P, K700A, and V702Q. In some embodiments, the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, D679N, E681N, K695D, L680E, and R685Q. In some embodiments, the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, K700A, V702Q, D679N. E681N, K695D, L680E, R685Q, and V170L. In some embodiments, the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, D679N, E681N K695D, L680E, R685Q, E682S, F678N, N688E, and V677T. In some embodiments, the ectodomain comprises tie amino acid substitutions E686L, R693V, V694L, Y690F, K700A, V702Q, D679N, E681N, K695D, L680E, R685Q, V701L, E682S, F678N, N688E, V677T, and D699K. In some embodiments, the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, D679H, E681N, K695D, L680E, R685Q, E682S, F678N, N688E, V677T, F687A, M684S, Q692S, and Y696R. In some embodiments, the ectodomain comprises the amino acid substitutions E686L, R693V, V694L, Y690F, K700A, V702E, D679H, E681N, K695D, L680E, R685Q, V701L, E682S, F678N, N688E, V677T, D699K, F687A, M684S, Q692S, and Y696R.

[0124] In some embodiments, the ectodomain comprises the amino acid substitution S367I or the amino acid substitution T374F. In some embodiments, the ectodomain comprises the amino acid substitutions S367I and T374F.

[0125] In some embodiments, the ectodomain comprises e, 2, 3, 4 or more amino acid substitutions selected from the group consisting of Q591F, Q591Y, S668A, Y218F, N220S, and V552L. In some embodiments, the ectodomain comprises the amino acid substitutions Q591F, S668A, and Y218F. In some embodiments, the ectodomain comprises the amino acid substitutions N220S and V552L. In some embodiments, the ectodomain comprises the amino acid substitutions Q591F, S668A, Y218F, N220S, and V552L. In some embodiments, the ectodomain comprises the amino acid substitutions Q591Y, 5668A, Y218F, N220S, and V552L. In some embodiments, the ectodomain comprises the amino acid substitutions Q591Y, N220S, and V552L. In some embodiments, the ectodomain comprises the amino acid substitutions Q591F, N220S, and V552L.

[0126] In some embodiments, the ectodomain comprises amino acid substitution E167T or E167S. In sonic embodiments, the ectodomain comprises 1, 2, 3, 4 or more amino acid substitutions selected from the group consisting of C246S, H157R, 1156H, R457S, R460S, W240N, and Y242T.

[0127] In some embodiments, the polypetide comprises any one combination of amino acid substitutions listed in Table 10.

[0128] In sonic embodiments, the polypetide comprises a polypeptide sequence at least 75%, 80%85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to a polypeptide sequence in Table 10 without the signal peptide, or an antigenic fragment thereof.

[0129] In some embodiments, the CMV gB protein adopts in a prefusion conformation in the absence of a fusion inhibitor, optionally N-{4-[({(]S)-1-[3,5-bis(trifluoromethyl)phenyl]ethyl}carbamothioyl) amino]phenyl}-1,3-thiazole-4-carboxamide.

[0130] In another aspect, the disclosure provides a polypeptide comprising an amino acid sequence at least 90%, 91%, 92%, 93%, 94%,95%, 96%, 97% 98%, or 99% identical to the amino acid sequence of SEQ ID NO:1 or SEQ ID NO:3 residues 91-702, or an antigenic fragment thereof, wherein residues 698-702 are optional if they do not include a mutation, SEQ ID NO:3 residue 456 is absent, and the polypeptide or antigenic fragment thereof comprises 1 or more combination of mutations relative to SEQ ID NO:1 or SEQ ID NO:3 residues 91-702 listed in Table 2, Table 3, or Table 4.

[0131] In some embodiments, the polypeptide or antigenic fragment thereof comprises 1 or more combination of mutations relative to SEQ ID NO:1 or SEQ ID NO:3 residues 91-702 listed in Table 2 or Table 3. In some embodiments, the polypeptide or antigenic fragment thereof comprises 1 or more combination of mutations relative to SEQ ID NO: 1 or SEQ ID NO:3 residues 91-702 listed in Table 2. In some embodiments, any polypeptide or antigenic fragment of the disclosure thereof further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, I1, 12, 13, 14, 15, 16, 17, 18, 19, 20, or more mutations relative to relative to SEQ ID NO: 1 or SEQ ID NO:3 residues 91-702 listed in Table 5. In some embodiments, any polypeptide or antigenic fragment of the disclosure thereof further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, or all 10 mutations relative to relative to SEQ ID NO:1 or SEQ ID NO:3 residues 91-702 listed in Table 6. In some embodiments, any polypeptide or antigenic fragment of the disclosure thereof further comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or all 12, relative to relative to SEQ ID NO.: or SEQ ID NO:3 residues 91-702 listed in Table 7. In some embodiments, any polypeptide or antigenic fragment of the disclosure thereof further comprises a combination of mutations relative to relative to SEQ ID NO: 1 or SEQ ID NO:3 residues 91-702 listed in Table 8.

[0132] In some embodiments, any polypeptide of the disclosure, comprising an amino acid sequence at least 90%, 91%, 92%, 93%, 94% 95%, 96%, 97% 98%, or 99% identical to the amino acid sequence selected from the sequences listed in Table 9, or an antigenic fragment thereof. In some embodiments, any polypeptide of the disclosure, comprising an amino acid sequence at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the amino acid sequence selected from the sequences listed in Table 10, or an antigenic fragment thereof. In some embodiments, any polypeptide of the disclosure further comprising any other functional domain as appropriate for an intended use, including but not limited to a secretion signal located at the N-terminus of the polypeptide, wherein the signal sequence may be any suitable signal sequence as appropriate for an intended use.

Signal Peptide

[0133] In some embodiments, the encoded polypeptide includes a peptide region that is a signal peptide. Signal peptides are well known in the art. The signal peptide may be the native signal peptide or can be replaced with another signal peptide. Signal peptides function to prompt a cell to translocate the protein, usually to the cellular membrane. The core of the signal peptide often contains a long stretch of hydrophobic amino acids (about 5-16 residues long) that has a tendency to form a single alpha-helix and is also referred to as the h-region. In addition, many signal peptides begin with a short positively charged stretch of amino acids, which may help to enforce proper topology of the polypeptide during translocation by what is known as the positive-inside rule. Because of its close location to the N-terminus it is called the n-region. At the end of the signal peptide there is typically a stretch of amino acids that is recognized and cleaved by signal peptidase.

[0134] In some embodiments, the polypeptide does not include a signal peptide.

[0135] Nonlimiting examples of signal peptides are provided below:

TABLE-US-00009 (SEQIDNO:32) MELLILKANAITTILTAVTFCFASG (SEQIDNO:33) MSWKVMIIISLLITPQHGL (SEQIDNO:34) MKAILVVLLYTFTTANA (SEQIDNO:35) MPISILLIITTMIMASHC (SEQIDNO:36) MFVFLVLLPLVSSQC (SEQIDNO:37) MVPQVLLFVPLLGFSLCFG

[0136] Illustrative sequences of fusions that include a signal peptide are provided in Table 9. In each case, the fusion protein is shown with a signal peptide that would be cleaved during secretion of the fusion protein. The signal peptide is shown in parentheses to identicate that sequence identity to each sequence should be calculated without the signal peptide. Alternative signal peptides may be substituted using routine methods known in the art.

TABLE-US-00010 TABLE9 SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:38 KOs_dII-dIII-pack1 SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST dII-dIII-pack1:S367I, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT T374F NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTANSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMREFNSYKQRVKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:39 KOs_Cpack1-short SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack1-short:E686L, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Y690F,R693V,V694L NRNASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSINLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:40 KOs_Cpack1-long SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack1-long:E686L,Y690F, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R693V,V694D,K700A,V702Q NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:41 KOs_Cpack2-short SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack2-short:D679N, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT L680E,E681N,R685Q, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER E686L,Y690F,R693V, ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM V694L,K695D TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSINLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVEN ENEIMQLENSFKQVLDYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:42 KOs_Cpack2-long SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack2-long:D679N,L680E, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT E681N,R685Q,E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Y690F,R693V,V694L, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM K695D,K700A,V701L,V702Q TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVEN ENEIMQLFNSFKQVLDYVEDALQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:43 KOs_Cpack3-short SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack3-short:V677T, NLTRETSNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT F678N,D679N,L680E, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER E681N,E682S,R685Q, ADSVISWDIQDEKNVICQLIFWEASERTIRSEAEDSYHESSAKM E686L,N688E,Y690F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY R693V,V694L,K695D GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTANSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTNN ENSIMQLFESFKQVLDYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:44 KOs_Cpack3-long SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack3-long:V677T,F678N, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT D679N,L680E,E681N, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER E682S,R685Q,E686L, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM N688E,Y690F,R693V, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V694L,K695D,D699K, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK K700A,V701L,V702Q SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNTNN ENSIMQLFESFKQVLDYVEKALQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:45 KOs_Cpack4-short SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack4-short:V677T, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT F678N,D679H,L680E, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHREVAFLER E681N,E682S,M684S, ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHESSAKM R685Q,E686L,F687A, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N688E,Y690F,Q692S, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK R693V,V694L,K695D,Y696R SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNINH ENSISQLAESFKSVLDRV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:46 KOs_Cpack4-long SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack4-long:V677T,F678N, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT D679H,L680E,E681N, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER E682S,M684S,R685Q, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM E686L,F687A,N688E, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Y690F,Q6925,R693V, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK V694L,K695D,Y696R, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA D699K,K700A,V701L,V702E EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNTNH ENSISQLAESFKSVLDRVEKALE SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:47 KOs_Cpack1-short_ SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dII-dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT dII-dIII-pack1: NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER S367I,T374F ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM Cpack1-short:E686L, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Y690F,R693V,V694L GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHITSAAHSRSGSVSQRVTSSQTV NO:48 KOs_Cpack1-long_ SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dII-dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1: NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER S367I,T374F ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM Cpack1-long: TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY E686L,Y690F, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTANSTK R693V,V694L,K700A,V702Q SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:49 KOs_Cpack2-short_ SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dII-dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM Cpack2-short:D679N, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY L680E,E681N,R685Q, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK E686L,Y690F,R693V, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA V694L,K695D EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVEN ENEIMQLFNSFKQVLDYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:50 KOs_Cpack2-long_ SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dII-dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM Cpack2-long:D679N,L680E, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY E681N,R685Q,E686L, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Y690F,R693V,V694L, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA K695D,K700A,V701L,V702Q EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVEN ENEIMQLENSFKQVLDYVEDALQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:51 KOs_Cpack3-short_ SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER pdII-dIII-ack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM Cpack3-short:V677T, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY F678N,D679N,L680E, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNITANSTK E681N,E682S,R685Q, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA E686L,N688E,Y690F, EAWCVDQRRILEVEKELSKINPSAILSAIYNKPIAARFMGDVLG R693V,V694L,K695D LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNINN ENSIMQLFESFKQVLDYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:52 KOs_Cpack3-long_ SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dII-dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM Cpack3-long:V677T,F678N, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY D679N,L680E,E681N, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK E682S,R685Q,E686L, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA N688E,Y690F,R693V, EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG V694L,K695D,D699K, LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ K700A,V701L,V702Q YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTNN ENSIMQLFESEKQVLDYVEKALQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:53 KOs_Cpack4-short_ SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dII-dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYARDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM Cpack4-short:V677T, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY F678N,D679H,L680E, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK E681N,E682S,M684S, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA R685Q,E686L,F687A, EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG N688E,Y690F,Q692S, LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ R693V,V694L,K695D,Y696R YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNINH ENSISQLAESFKSVLDRV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:54 KOs_Cpack4-long_ SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dII-dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER T374F ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM Cpack4-long:V677T,F678N, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY D679H,L680E,E681N, GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK E682S,M684S,R685Q, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA E686L,F687A,N688E, EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG Y690F,Q692S,R693V, LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ V6941,K695D,Y696R, YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR D699K,K700A,V701L,V702E MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNINH ENSISQLAESFKSVLDRVEKALE SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:55 KOs_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack1:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVFD LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:56 KOs_Cpack1-long_ SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dI-dIV-dV-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K700A,V702Q ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack1:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTANSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:57 KOs_Cpack1-short_Disulf1 SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVEVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST Cpack1-short:E686L, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Y690F,R693V,V694L NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:58 KOs_Cpack1-long_Disulf1 SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST Cpack1-long:E686L,Y690F, NLTRETSNINCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R693V,V694L,K700A,V702Q NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:59 KOs_Cpack1-short_Disulf2 SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVEVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST Cpack1-short:E686L, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Y690F,R693V,V694L NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHREVAFLER Disulf2:D217C,Y589C ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTANSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:60 KOs_Cpack1-long_Disulf2 SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Bg-KOs:I156H,H157R, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR W240N,Y242T,C246S, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R457S,R460S TVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST Cpack1-long:E6861,Y690F, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R693V,V694L,K700A,V702Q NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf2:D217C,Y589C ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:61 KOs_Cpack1-short_ SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dI-dIV-dV-pack1_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack1:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMPLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:62 KOs_Cpack1-long_dI-dIV-dV- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER pack1_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K700A,V702Q ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM dI-dIV-dV-pack1:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNITHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:63 KOs_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack1_Disulf2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack1:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Disulf2:D217C,Y589C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:64 KOs_Cpack1-long_dI-dIV-dV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_Disulf2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K700A,V702Q ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack1:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Disulf2:D217C,Y589C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVFD LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:65 KOs_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER dV-pack1N165glyc NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTTYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack1:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTANSTK N165glyc:E167T SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRILEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:66 KOs_Cpack1-long_dI-dIV-dV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_N165glyc NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTTYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLER R693V,V694L,K700A,V702Q ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHESSAKM dI-dIV-dV-pack1:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK N165glyc:E167T SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:67 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER T374F ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM Cpack1-short:E686L, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Y690F,R693V,V694L GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSINLTHNSTK dI-dIV-dV-pack1:Y218F, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA Q591F,S668A EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANSSYVF YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:68 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM Cpack1-long:E686L,Y690F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY R693V,V694L,K700A,V702Q GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK dI-dIV-dV-pack1:Y218F, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA Q591F,S668A EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:69 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM Cpack1-short:E686L, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Y690F,R693V,V694L GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:70 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKIMQLMPDCYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER T374F ADSVISWDIQDEKNVICQLIFWEASERTIRSEAEDSYHFISAKM Cpack1-long:E686L,Y690F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY R693V,V694L,K700A,V702Q GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHITSAAHSRSGSVSQRVTSSQTV NO:71 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGIDLIRFER pack1_Disulf2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM Cpack1-short:E686L, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Y690F,R693V,V694L GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTANSTK Disulf2:D217C,Y589C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO: KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_Disulf2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM Cpack1-long:E686L,Y690F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY R693V,V694L,K700A,V702Q GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Disulf2:D217C,Y589C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:73 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack1_Disulf1 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dII-dIII-pack1:S367I, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM T374F TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Cpack1-short:E686L, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Y690F,R693V,V694L SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack1:Y218F, EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG Q591F,S668A LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE Disulf1:D217C,S587C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:74 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack1_Disulf1 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVEVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dII-dIII-pack1:S367I, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM T374F TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Cpack1-long:E686L,Y690F, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK R693V,V694L,K700A,V702Q SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack1:Y218F, EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG Q591F,S668A LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE Disulf1:D217C,S587C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:75 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack1_Disulf2 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dII-dIII-pack1:S367I, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM T374F TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Cpack1-short:E686L, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Y690F,R693V,V694L SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack1:Y218F, EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG Q591F,S668A LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVE Disulf2:D217C,Y589C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:76 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack1_Disulf2 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVEVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dII-dIII-pack1:S367I, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM T374F TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Cpack1-long:E686L,Y690F, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSINLTHNSTK R693V,V694L,K700A,V702Q SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack1:Y218F, EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG Q591F,S668A LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVF Disulf2:D217C,Y589C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:77 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR pack1_N165glyc SYAYHRTTYLLGSNTTYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dII-dIII-pack1:S367I, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM T374F TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Cpack1-short:E686L, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Y690F,R693V,V694L SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack1:Y218F, EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG Q591F,S668A LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE N165glyc:E167T YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:78 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack1_N165glyc SYAYHRTTYLLGSNTTYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dII-dIII-pack1:3367I, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM T374F TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Cpack1-long:E686L,Y690F, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK R693V,V694L,K700A,V702Q SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack1:Y218F, EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG Q591F,S668A LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE N165glyc:E167T YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:79 KOs_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM dI-dIV-dV-pack2:N220S, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V5521 GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:80 KOs_Cpack1-long_dI-dIV-dV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K700A,V7020 ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack2:N220S, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V552L GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:81 KOs_Cpackl-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack3 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack3:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N220S,V552L,Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:82 KOs_Cpack1-long_dI-dIV-dV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack3 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K700A,V702Q ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM dI-dIV-dV-pack3:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N220S,V552L,Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:83 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV-pack2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM Cpack1-short:E686L, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Y690F,R693V,V694L GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK dI-dIV-dV-pack2:N220S, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA V552L EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVFD LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:84 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV-pack2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHREVAFLER T374F ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM Cpack1-long:E686L,Y690F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY R693V,V694L,K700A,V702Q GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTANSTK dI-dIV-dV-pack2:N220S, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA V552L EAWCVDQRRILEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:85 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV-pack3 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM Cpack1-short:E686L, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Y690F,R693V,V694L GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK dI-dIV-dV-pack3:Y218F, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA N220S,V552L,Q591F,S668A EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:86 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV-pack3 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM Cpack1-long:E686L,Y690F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY R693V,V694L,K700A,V702Q GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK dI-dIV-dV-pack3:Y218F, SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA N220S,V552L,Q591F,S668A EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:87 KOs_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack3_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHESSAKM dI-dIV-dV-pack3:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N220S,V552D,Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:88 KOs_Cpack1-long_dI-dIV-dV- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGIDLIRFER pack3_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCESSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K700A,V702Q ADSVISWDIQDEKNVICQLIFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack3:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N220S,V552L,Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Disulf1:D217C,S587C SSIDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRILEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:89 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack3_Disulf1 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVEVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Cpack1-short:E686L, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM Y690F,R693V,V694L TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY dII-dIII-pack1:S367I, GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK T374F SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack3:Y218F, EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG N220S,V552L,Q591F,S668A LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVF Disulf1:D217C,S587C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:90 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack3_Disulf1 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVEVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Cpack1-long:E686L,Y690F, ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM R693V,V694D,K700A,V702Q TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY dII-dIII-pack1:S367I, GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK T374F SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack3:Y218F, EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG N220S,V552L,Q591F,S668A LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE Disulf1:D217C,S587C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:91 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTFRR pack4_Disulf1 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVEVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Cpack1-short:E686L, ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM Y690F,R693V,V694L TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY dII-dIII-pack1:S367I, GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK T374F SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack4:Y218F, EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG N220S,V552L,Q591Y,S668A LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY Disulf1:D217C,S587C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:92 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack4_Disulf1 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVEVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER Cpack1-long:E686L,Y690F, ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM R693V,V694L,K700A,V702Q TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY dII-dIII-pack1:S367I, GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK T374F SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack4:Y218F, EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG N220S,V552L,Q591Y,S668A LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANCSYVY Disulf1:D217C,S587C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:93 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack5_Disulf1 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVEVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Cpack1-short:E686L, ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM Y690F,R693V,V694L TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY dII-dIII-pack1:S367I, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK T374F SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack5:N220S, EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG V5521,Q591Y LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY Disulf1:D217C,S587C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:94 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack5_Disulf1 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVEVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER Cpack1-long:E686L,Y690F, ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM R693V,V694D,K700A,V702Q TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY dII-dIII-pack1:S367I, GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK T374F SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack5:N220S, EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG V552L,Q591Y LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANCSYVY Disulf1:D217C,S587C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:95 KOs_Cpack1-short_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack6_Disulf1 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Cpack1-short:E686L, ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM Y690F,R693V,V694L TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY dII-dIII-pack1:S367I, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK T374F SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack6:N220S, EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG V5521,Q591F LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVF Disulf1:D217C,S587C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHASSHTTSAAHSRSGSVSQRVTSSQTV NO:96 KOs_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack6_Disulf1 SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Bg-KOs:I156H,H157R, TVEVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST W240N,Y242T,C246S, NLTRETSNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT R457S,R460S NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER Cpack1-long:E686L,Y690F, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM R693V,V694D,K700A,V702Q TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY dII-dIII-pack1:S367I, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK T374F SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA dI-dIV-dV-pack6:N220S, EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG V552L,Q591F LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANCSYVE Disulf1:D217C,S587C YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVFD LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:97 KOs_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack4_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKIMQLMPDCFSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHESSAKM dI-dIV-dV-pack4:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N220S,V552L,Q591Y,S668A GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHITSAAHSRSGSVSQRVTSSQTV NO:98 KOs_Cpack1-long_dI-dIV-dV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack4_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K700A,V702Q ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack4:Y218F, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N220S,V552L,Q591Y,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:99 KOs_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER dV-pack5_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack5:N220S, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V552L,Q591Y GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSINLTHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:100 KOs_Cpack1-long_dI-dIV-dV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack5_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694D,K700A,V702Q ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack5:N220S, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V552L,Q591Y GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSINLTHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:101 KOs_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack6_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVEVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack6:N220S, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V552L,Q591F GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVF YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVFD LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N-term_Bg- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:102 KOs_Cpack1-long_dI-dIV-dV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGIDLIRFER pack6_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR Bg-KOs:I156H,H157R, SYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG W240N,Y242T,C246S, TVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST R457S,R460S NLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K700A,V702Q ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFSSAKM dI-dIV-dV-pack6:N220S, TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V5521,Q591F GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNSTK Disulf1:D217C,S587C SSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N-term_dII- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:103 dIII-pack1 SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dII-dIII-pack1:S367I, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR T374F SYAYINTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSENLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMREFNSYKQRVKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:104 term_Cpack1-short SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGIDLIRFER Cpack1-short:E686L, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Y690F,R693V,V694L SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:105 term_Cpack1-long SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGIDLIRFER Cpack1-long:E686L,Y690F, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR R693V,V694L,K700A,V702Q SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDILRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:106 term_Cpack2-short SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Cpack2-short:D679N, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR L680E,E681N,R685Q, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG E686L,Y690F,R693V, TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST V694L,K695D WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVEN ENEIMQLFNSFKQVLDYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:107 term_Cpack2-long SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Cpack2-long:D679N,L680E, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR E681N,R685Q,E686L, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L, TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST K695D,K700A,V701L,V702Q WLYRETCNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRIVAFLER ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVEN ENEIMQLENSFKQVLDYVEDALQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:108 term_Cpack3-short SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER Cpack3-short:V677T, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR F678N,D679N,L680E, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG E681N,E682S,R685Q, TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST E686L,N688E,Y690F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R693V,V694L,K695D NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNINN ENSIMQLFESFKQVLDYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:109 term_Cpack3-long SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER Cpack3-long:V677T,F678N, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR D679N,L680E,E681N, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG E682S,R685Q,E686L, TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST N688E,Y690F,R693V, WLYRETCNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT V694L,K695D,D699K, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER K700A,V701L,V702Q ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTNN ENSIMQLFESFKQVLDYVEKALQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:110 term_Cpack4-short SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Cpack4-short:V677T, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR F678N,D679H,L680E, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG E681N,E682S,M684S, TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST R685Q,E686L,F687A, WLYRETCNINCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT N688E,Y690F,Q692S, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K695D,Y696R ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNINH ENSISQLAESFKSVLDRV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:111 term_Cpack4-long SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER Cpack4-long:V677T,F678N, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR D679H,L680E,E681N, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG E682S,M684S,R685Q, TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST E686L,F687A,N688E, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Y690F,Q692S,R693V, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER V694L,K695D,Y696R, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM D699K,K700A,V701L,V702E TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRILEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNINH ENSISQLAESFKSVLDRVEKALE SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:112 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack1-short:E686L, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Y690F,R693V,V694L NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:113 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYINTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack1-long:E686L,Y690F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R693V,V694L,K700A,V702Q NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVFD LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHITSAAHSRSGSVSQRVTSSQTV NO:114 term_Cpack2-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack2-short:D679N, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT L680E,E681N,R685Q, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER E686L,Y690F,R693V, ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM V694L,K695D TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVFN ENEIMQLFNSFKQVLDYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:115 term_Cpack2-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack2-long:D679N,L680E, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT E681N,R685Q,E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Y690F,R693V,V694L, ADSVISWDIQDEKNVICQLIFWEASERTIRSEAEDSYHFISAKM K695D,K700A,V701L,V702Q TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTERGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVEN ENEIMQLENSFKQVLDYVEDALQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:116 term_Cpack3-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack3-short:V677T, WLYRETCNINCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT F678N,D679N,L680E, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER E681N,E682S,R685Q, ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM E686L,N688E,Y690F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY R693V,V694L,K695D GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTNN ENSIMQLFESEKQVLDYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:117 termCpack3-longdII-dIII- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack3-long:V677T,F678N, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT D679N,L680E,E681N, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER E682S,R685Q,E686L, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM N688E,Y690F,R693V, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V694L,K695D,D699K, GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK K700A,V701L,V702Q RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNTNN ENSIMQLFESFKQVLDYVEKALQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:118 term_Cpack4-short_dII- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGIDLIRFER dIII-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack4-short:V677T, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT F678N,D679H,L680E, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER E681N,E682S,M684S, ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM R685Q,E686L,F687A, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N688E,Y690F,Q692S, GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK R693V,V694L,K695D,Y696R RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRILEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNINH ENSISQLAESFKSVLDRV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:119 term_Cpack4-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRGST Cpack4-long:V677T,F678N, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT D679H,L680E,E681N, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER E682S,M684S,R685Q, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM E686L,F687A,N688E, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Y690F,Q692S,R693V, GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK V694L,K695D,Y696R, RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA D699K,K700A,V701L,V702E EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTNH ENSISQLAESFKSVLDRVEKALE SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHASSHTTSAAHSRSGSVSQRVTSSQTV NO:120 term_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-short:E686L, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L TVEVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack1:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVF YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVFD LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:121 term_Cpack1-long_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGIDLIRFER dV-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-long:E686L,Y690F, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R693V,V694L,K700A,V702Q TVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack1:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVF YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:122 term_Cpack1-short_Disulf1 SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Cpack1-short:E686L, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Y690F,R693V,V694L SYAYINTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Disulf1:D217C,S587C TVEVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANCSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:123 term_Cpack1-long_Disulf1 SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Cpack1-long:E686L,Y690F, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR R693V,V694L,K700A,V702Q SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Disulf1:D217C,S587C TVEVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:124 term_Cpack1-short_Disulf2 SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGIDLIRFER Cpack1-short:E686L, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Y690F,R693V,V694L SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Disulf2:D217C,Y589C TVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSINLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTERGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:125 term_Cpack1-long_Disulf2 SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER Cpack1-long:E686L,Y690F, NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR R693V,V694L,K700A,V702Q SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Disulf2:D217C,Y589C TVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANSSCVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:126 term_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack1_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-short:E686L, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L TVEVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack1:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVF YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:127 termCpacki-long_dI-dIV- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER dV-pack1_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-long:E686L,Y690F, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R693V,V694L,K700A,V702Q TVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack1:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:128 term_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER dV-pack1_Disulf2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR Cpack1-short:E686L, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L TVEVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack1:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf2:D217C,Y589C ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSINLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:129 term_Cpack1-long_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack1_Disulf2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-long:E686L,Y690F, SYAYINTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R693V,V694L,K700A,V702Q TVEVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack1:Y218F, WLYRETCNINCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf2:D217C,Y589C ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVF YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVFD LEEIMPLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:130 term_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER dV-pack1_N165glyc NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-short:E686L, SYAYIHTTYLLGSNTTYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L TVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack1:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER N165glyc:E167T ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTERGYINRALAQIA EAWCVDQRRILEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:131 term_Cpack1-long_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack1_N165glyc NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR Cpack1-long:E686L,Y690F, SYAYIHTTYLLGSNTTYVAPPMWEIHHINSHSQCYSSYSRVIAG R693V,V694L,K700A,V702Q TVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack1:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLER N165glyc:E167T ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:132 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_dI-dIV-dV-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTFRR dII-dIII-pack1:S367I, SYAYINTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRGST Cpack1-short:E686L, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Y690F,R693V,V694L NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dI-dIV-dV-pack1:Y218F, ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM Q591F,S668A TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVFD LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:133 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV-pack1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRGST Cpack1-long:E686L,Y690F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R693V,V694L,K700A,V702Q NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dI-dIV-dV-pack1:Y218F, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM Q591F,S668A TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:134 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST Cpack1-short:E686L, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Y690F,R693V,V694L NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:135 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVEVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST Cpack1-long:E686L,Y690F, WLYRETCNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT R693V,V694L,K700A,V702Q NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,3587C ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:136 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_Disulf2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST Cpack1-short:E686L, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Y690F,R693V,V694L NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf2:D217C,Y589C ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:137 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_Disulf2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRGST Cpack1-long:E686L,Y690F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R693V,V694L,K700A,V702Q NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf2:D217C,Y589C ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSINLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:138 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR pack1_Disulf1 SYAYIRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG dII-dIII-pack1:S367I, TVEVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST T374F WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack1:Y218F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK Disulf1:D217C,S587C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHASSHITSAAHSRSGSVSQRVTSSQTV NO:139 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack1_Disulf1 SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG dII-dIII-pack1:S367I, TVEVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST T374F WLYRETCNINCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K700A,V702Q ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack1:Y218F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK Disulf1:D217C,S587C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANCSYVF YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVFD LEEIMPLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:140 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack1_Disulf2 SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG dII-dIII-pack1:S367I, TVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST T374F WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack1:Y218F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK Disulf2:D217C,Y589C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSCVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:141 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack1_Disulf2 SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG dII-dIII-pack1:S367I, TVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRGST T374F WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694D,K700A,V702Q ADSVISWDIQDEKNVICQLIFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack1:Y218F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK Disulf2:D217C,Y589C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANSSCVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:142 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack1_N165glyc SYAYINTTYLLGSNTTYVAPPMWEIHHINSHSQCYSSYSRVIAG dII-dIII-pack1:S367I, TVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRGST T374F WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-short:E686L, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Y690F,R693V,V694L ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack1:Y218F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK N165glyc:E167T RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVF YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:143 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGIDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack1_N165glyc SYAYIHTTYLLGSNTTYVAPPMWEIHHINSHSQCYSSYSRVIAG dII-dIII-pack1:S367I, TVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRGST T374F WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Cpack1-long:E686L,Y690F, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER R693V,V694L,K700A,V702Q ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack1:Y218F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK N165glyc:E167T RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCVTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:144 term_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-short:E686L, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L TVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack2:N220S, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT V552L NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:145 term_Cpack1-long_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-long:E686L,Y690F, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R693V,V694L,K700A,V702Q TVEVAYHRDSYENKIMQLMPDDYSSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack2:N220S, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT V552L NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVED LEEIMPLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHITSAAHSRSGSVSQRVTSSQTV NO:146 term_Cpackl-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack3 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-short:E686L, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L TVFVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack3:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT N220S,V552L,Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:147 term_Cpack1-long_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack3 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR Cpack1-long:E686L,Y690F, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R693V,V694L,K700A,V702Q TVEVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack3:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT N220S,V552L,Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSENLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:148 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGIDLIRFER dIII-pack1_dI-dIV-dV-pack2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVEVAYHRDSYENKTMQLMPDDYSSTRSTRYVTVKDQWHSRGST Cpack1-short:E686L, WLYRETCNINCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Y690F,R693V,V694L NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dI-dIV-dV-pack2:N220S, ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM V552L TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVFD LEEIMPLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:149 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV-pack2 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRGST Cpack1-long:E686L,Y690F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R693V,V694L,K700A,V702Q NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dI-dIV-dV-pack2:N220S, ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM V552L TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTANRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTERGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVQ YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:150 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_dI-dIV-dV-pack3 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR dII-dIII-pack1:S367I, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVFVAYHRDSYENKTMQLMPDDFSSTHSTRYVTVKDQWHSRGST Cpack1-short:E686L, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT Y690F,R693V,V694L NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dI-dIV-dV-pack3:Y218F, ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM N220S,V552L,Q591F,S668A TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:151 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV-pack3 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTFRR dII-dIII-pack1:S367I, SYAYINTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG T374F TVEVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRGST Cpack1-long:E686L,Y690F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT R693V,V694L,K700A,V702Q NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER dI-dIV-dV-pack3:Y218F, ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM N220S,V552L,Q591F,S668A TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRIK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANSSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:152 term_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack3_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-short:E686L, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L TVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack3:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT N220S,V552L,Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYV SEQID GB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:153 term_Cpack1-long_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack3_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-long:E686L,Y690F, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R693V,V694L,K700A,V702Q TVEVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack3:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT N220S,V552L,Q591F,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:154 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack3_Disulf1 SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Cpack1-short:E686L, TVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST Y690F,R693V,V694L WLYRETCNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack3:Y218F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N220S,V552L,Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK Disulf1:D217C,S587C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVF YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHETSAAHSRSGSVSQRVTSSQTV NO:155 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack3_Disulf1 SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Cpack1-long:E686L,Y690F, TVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST R693V,V694L,K700A,V702Q WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack3:Y218F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N220S,V552L,Q591F,S668A GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK Disulf1:D217C,S587C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:156 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack4_Disulf1 SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Cpack1-short:E686L, TVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST Y690F,R693V,V694L WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack4:Y218F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N220S,V552L,Q591Y,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSINLTHNRTK Disulf1:D217C,S587C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:157 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR pack4_Disulf1 SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Cpack1-long:E686L,Y690F, TVEVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST R693V,V694L,K700A,V702Q WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack4:Y218F, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY N220S,V552L,Q591Y,S668A GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK Disulf1:D217C,S587C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHASSHTTSAAHSRSGSVSQRVTSSQTV NO:158 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack5_Disulf1 SYAYINTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Cpack1-short:E686L, TVEVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST Y690F,R693V,V694L WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack5:N220S, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V552L,Q591Y GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK Disulf1:D217C,S587C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVFD LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:159 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTFRR pack5_Disulf1 SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Cpack1-long:E686L,Y690F, TVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST R693V,V694D,K700A,V7020 WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack5:N220S, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V552L,Q591Y GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK Disulf1:D217C,S587C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:160 term_Cpack1-short_dII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dIII-pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack6_Disulf1 SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Cpack1-short:E686L, TVEVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST Y690F,R693V,V694L WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack6:N220S, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V552L,Q591F GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK Disulf1:D217C,S587C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:161 term_Cpack1-long_dII-dIII- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER pack1_dI-dIV-dV- NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR pack6_Disulf1 SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Cpack1-long:E686L,Y690F, TVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST R693V,V694L,K700A,V702Q WLYRETCNINCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT dII-dIII-pack1:S367I, NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER T374F ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKM dI-dIV-dV-pack6:N220S, TAFFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY V5521,Q591F GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK Disulf1:D217C,S587C RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID GB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:162 term_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGIDLIRFER dV-pack4_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-short:E686L, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L TVEVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack4:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT N220S,V552L,Q591Y,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVICQLIFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVFD LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:163 term_Cpack1-long_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack4_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTFRR Cpack1-long:E686L,Y690F, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R693V,V694L,K700A,V702Q TVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack4:Y218F, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT N220S,V552L,Q591Y,S668A NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVED LEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:164 term_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack5_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-short:E686L, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L TVEVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack5:N220S, WLYRETCNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGT V552L,Q591Y NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLENSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:165 term_Cpack1-long_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack5_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-long:E686L,Y690F, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R693V,V694L,K700A,V702Q TVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack5:N220S, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT V552L,Q591Y NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHESSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVY YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLEKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ SEQID GB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:166 term_Cpack1-short_dI-dIV- SHGVNETIYNTTLKYGDVVGVNITKYPYRVCSMAQGTDLIRFER dV-pack6_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTEKVRVYQKVLTERR Cpack1-short:E686L, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG Y690F,R693V,V694L TVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack6:N220S, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT V5521,Q591F NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSIVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVFKELSKINPSAILSAIYNKPIAAREMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIENFANCSYVE YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVED LEEIMRLFNSFKQVLKYV SEQID gB-Towne-ext-N- GAAVSSSSTRGTSATHSHHSSHTTSAAHSRSGSVSQRVTSSQTV NO:167 term_Cpack1-long_dI-dIV- SHGVNETIYNTTLKYGDVVGVNTTKYPYRVCSMAQGTDLIRFER dV-pack6_Disulf1 NIVCTSMKPINEDLDEGIMVVYKRNIVAHTFKVRVYQKVLTERR Cpack1-long:E686L,Y690F, SYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSSYSRVIAG R693V,V694L,K700A,V702Q TVEVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRGST dI-dIV-dV-pack6:N220S, WLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGT V552L,Q591F NRNASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLER Disulf1:D217C,S587C ADSVISWDIQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKM TATFLSKKQEVNMSDSALDCVRDEAINKLQQIFNTSYNQTYEKY GNVSVFETTGGLVVFWQGIKQKSLVELERLANRSSLNLTHNRTK RSTDGNNATHLSNMESVHNLVYAQLQFTYDTLRGYINRALAQIA EAWCVDQRRTLEVEKELSKINPSAILSAIYNKPIAARFMGDVLG LASCLTINQTSVKVLRDMNVKESPGRCYSRPVVIFNFANCSYVF YGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYVDYLFKR MIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVFD LEEIMRLFNSFKQVLKYVEDAVQ

Timerization Domain

[0137] In some embodiments, the encoded polypeptide includes a heterologous trimerization domain. In some embodiments, the trimerization domain is at the C-terminus of the viral protein monomer. In some embodiments, the trimerization domain is a Foldon. The sequence of a representative FoldOn is as follows:

TABLE-US-00011 (SEQIDNO:308) GYIPEAPRDGQAYVRKDGEWVLLSTF

[0138] In some embodiments, the trimerization domain is a GCN4 coiled-coil domain. The sequence of a representative GCN4 coiled-coil domain is as follows:

TABLE-US-00012 (SEQIDNO:309) IEDKIEEILSKIYHIENEIARIKKLI

[0139] In some embodiments, the assembly domain is expressed as part of a fusion protein that comprises a viral protein monomer, a linker, and the assembly domain. Illustrative sequences of fusion proteins are provided in Table 10. In each case, in further embodiments, the assembly domain is any one of the assembly domains provided in Table 10, or another assembly domain.

TABLE-US-00013 TABLE10 SEQID gB-Towne_Bg-KOs_dII- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:168 dIII-pack1 VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Bg-KOs:I156H,H157R, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG W240N,Y242T,C246S, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R457S,R460S ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD dII-dIII-pack1:S367I, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM T374F SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMREENSYKQRVKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:169 KOs_Cpack1-short VRVYQKVLIFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Bg-KOs:I156H,H157R, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG W240N,Y242T,C246S, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R457S,R460S ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Cpack1-short:E686L, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Y690F,R693V,V694L SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSIVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:170 KOs_Cpack1-long VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Bg-KOs:I156H,H157R, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG W240N,Y242T,C246S, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R457S,R460S ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Cpack1-long:E686L, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Y690F,R693V,V694L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG K700A,V702Q IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:171 KOs_Cpack2-short VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Bg-KOs:I156H,H157R, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG W240N,Y242T,C246S, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R457S,R460S ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Cpack2-short:D679N, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM L680E,E681N,R685Q, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG E686L,Y690F,R693V, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA V694L,K695D QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF NENEIMQLENSFKQVLDYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:172 KOs_Cpack2-long VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Bg-KOs:I156H,H157R, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG W240N,Y242T,C246S, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R457S,R460S ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Cpack2-long:D679N, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATELSKKQEVNM L680E,E681N,R685Q, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG E686L,Y690F,R693V, IKQKSIVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA V694L,K695D,K700A, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA V701L,V702Q IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE NENEIMQLENSFKQVLDYVEDALQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:173 KOs_Cpack3-short VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Bg-KOs:I156H,H157R, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG W240N,Y242T,C246S, STNLTRETSNINCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R457S,R460S ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Cpack3-short:V677T, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM F678N,D679N,L680E, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG E681N,E682S,R685Q, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA E686L,N688E,Y690F, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA R693V,V694L,K695D IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTN NENSIMQLFESFKQVLDYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:174 KOs_Cpack3-long VRVYQKVLTFRRSYAYHRITYLLGSNTEYVAPPMWEIHHINSHSQCYSS Bg-KOs:I156H,H157R, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG W240N,Y242T,C246S, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R457S,R460S ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Cpack3-long:V677T, IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM F678N,D679N,L680E, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG E681N,E682S,R685Q, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA E686L,N688E,Y690F, QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA R693V,V694L,K695D, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV D699K,K700A,V701L, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV V702Q DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNIN NENSIMQLFESFKQVLDYVEKALQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:175 KOs_Cpack4-short VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Bg-KOs:I156H,H157R, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVIVKDQWHSRG W240N,Y242T,C246S, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R457S,R460S ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Cpack4-short:V677T, IQDEKNVTCQLIFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM F678N,D679H,L680E, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG E681N,E682S,M684S, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA R685Q,E686L,F687A, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA N688E,Y690F,Q692S, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV R693V,V694L,K695D, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Y696R DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTN HENSISQLAESFKSVLDRV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:176 KOs_Cpack4-long VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Bg-KOs:I156H,H157R, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG W240N,Y242T,C246S, STNLIRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R457S,R460S ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Cpack4-long:V677T, IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMIATFLSKKQEVNM F678N,D679H,L680E, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG E681N,E682S,M684S, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA R685Q,E686L,F687A, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA N688E,Y690F,Q692S, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV R693V,V694L,K695D, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Y696R,D699K,K700A, DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNIN V701L,V702E HENSISQLAESFKSVLDRVEKALE SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:177 KOs_Cpack1-short_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1 YSRVIAGTVFVAYHRDSYENKIMQLMPDDYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSIVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Cpack1-short:E686L, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y690F,R693V,V694L IYNKPIAARFMGDVLGLASCVTINQTSVKVERDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:178 KOs_Cpack1-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1 YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRENSALETARLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Cpack1-long:E686L, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y690F,R693V,V694L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV K700A,V702Q IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:179 KOs_Cpack2-short_dII- VRVYQKVLIFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1 YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRENSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA Cpack2-short:D679N, QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA L680E,E681N,R685Q, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV E686L,Y690F,R693V, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV V694L,K695D DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE NENEIMQLENSFKQVLDYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:180 KOs_Cpack2-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1 YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNSTKSSIDGNNATHLSNMESVHNLVYA Cpack2-long:D679N, QLQFTYDELRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA L680E,E681N,R685Q, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV E686L,Y690F,R693V, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV V694L,K695D,K700A, DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF V701L,V702Q NENEIMQLENSFKQVLDYVEDALQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCISMKPINEDLDEGIMVVYKRNIVAHTEK NO:181 KOs_Cpack3-short_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1 YSRVIAGTVFVAYHRDSYENKIMQLMPDDYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFELSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSIVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Cpack3-short:V677T, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA F678N,D679N,L680E, IYNKPIAARFMGDVLGLASCVTINQTSVKVERDMNVKESPGRCYSRPVV E681N,E682S,R685Q, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV E686L,N688E,Y690F, DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTN R693V,V694L,K695D NENSIMQLFESFKQVLDYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:182 KOs_Cpack3-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1 YSRVIAGTVFVAYHRDSYENKIMQLMPDDYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Cpack3-long:V677T, QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA F678N,D679N,L680E, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV E681N,E682S,R6850, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV E686L,N688E,Y690F, DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTN R693V,V694L,K695D, NENSIMQLFESFKQVLDYVEKALQ D699K,K700A,V701L, V702Q SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:183 KOs_Cpack4-short_dII- VRVYQKVLIFRRSYAYHRITYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1 YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETARLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFELSKKQEVNM dII-dIII-pack1:3367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Cpack4-short:V677T, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA F678N,D679H,L680E, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV E681N,E682S,M684S, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV R685Q,E686L,F687A, DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNIN N688E,Y690F,Q692S, HENSISQLAESFKSVLDRV R693V,V694L,K695D, Y696R SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:184 KOs_Cpack4-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1 YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Cpack4-long:V677T, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA F678N,D679H,L680E, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV E681N,E682S,M684S, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV R685Q,E686L,F687A, DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNIN N688E,Y690F,Q692S, HENSISQLAESFKSVLDRVEKALE R693V,V694L,K695D, Y696R,D699K,K700A, V701L,V702E SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:185 KOs_Cpack1-short_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack1 YSRVIAGTVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSIVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack1: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,Q591F,S668A IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:186 KOs_Cpackl-long_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack1 YSRVIAGTVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack1: IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,Q591F,S668A IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:187 KOs_Cpack1- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS short_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Disulf1:D217C,S587C QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:188 KOs_Cpack1- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS long_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA Disulf1:D217C,S587C IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:189 KOs_Cpack1- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS short_Disulf2 YSRVIAGTVFVAYHRDSYENKIMQLMPDCYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R; STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLIFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSIVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Disulf2:D217C,Y589C QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVERDMNVKESPGRCYSRPVV IFNFANSSCVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:190 KOs_Cpack1- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS long_Disulf2 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Disulf2:D217C,Y589C IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSCVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:191 KOs_Cpack1-short_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack1_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRENSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack1: QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,Q591F,S668A IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Disulf1:D217C,S587C IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:192 KOs_Cpack1-long_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack1_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINPALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack1: IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,Q591F,S668A IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf1:D217C,S587C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:193 KOs_Cpack1-short_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack1Disulf2 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack1: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,Q591F,S668A IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Disulf2:D217C,Y589C IFNFANSSCVFYGQLGEDNEILLGNARTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:194 KOs_Cpack1-long_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack1_Disulf2 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA dI-dIV-dV-pack1: IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,Q591F,S668A IFNFANSSCVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf2:D217C,Y589C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:195 KOs_Cpack1-short_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNITYVAPPMWEIHHINSHSQCYSS dIV-dV-pack1_N165glyc YSRVIAGTVFVAYHRDSYENKIMQLMPDDESNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack1: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,Q591F,S668A IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV N165glyc:E167T IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:196 KOs_Cpackl-long_dI- VRVYQKVLIFRRSYAYHRTTYLLGSNTTYVAPPMWEIHHINSHSQCYSS dIV-dV-pack1_N165glyc YSRVIAGTVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATELSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack1: IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,Q591F,S668A IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV N165glyc:E167T DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:197 KOs_Cpack1-short_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRG pack1 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpack1-short:E686L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV dI-dIV-dV-pack1: DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF Y218F,Q591F,S668A DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:198 KOs_Cpack1-long_dII- VRVYQKVLTFRRSYAYHRITYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRG pack1 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpack1-long:E686L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L, IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV K700A,V702Q DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF dI-dIV-dV-pack1: DLEEIMRLENSFKQVLKYVEDAVQ Y218F,Q591F,S668A SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:199 KOs_Cpack1-short_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_Disulf1 YSRVIAGIVEVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Cpack1-short:E686L, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y690F,R693V,V694L IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Disulf1:D217C,S587C IFNFANCSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:200 KosCpack1-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSIVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Cpack1-long:E686L, QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y690F,R693V,V694L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV K700A,V702Q IFNFANCSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE Disulf1:D217C,S587C DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:201 KOs_Cpack1-short_dII- VRVYQKVLTFRRSYAYHRITYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_Disulf2 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Cpack1-short:E686L, QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA Y690F,R693V,V694L IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Disulf2:D217C,Y589C IFNFANSSCVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:202 KOs_Cpack1-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_Disulf2 YSRVIAGTVEVAYHRDSYENKIMQLMPDCYSNTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Cpack1-long:E686L, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y690F,R693V,V694L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV K700A,V702Q IFNFANSSCVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf2:D217C,Y589C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCISMKPINEDLDEGIMVVYKRNIVAHTFK NO:203 KOs_Cpack1-short_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG pack1_Disulf1 SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA Cpack1-short:E686L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV dI-dIV-dV-pack1: DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE Y218F,Q591F,S668A DLEEIMRLENSFKQVLKYV Disulf1:D217C,S587C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:204 KOs_Cpack1-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG pack1_Disulf1 SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFELSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSEVELERLANRSSINLTANSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpack1-long:B686L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L, IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV K700A,V702Q DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE dI-dIV-dV-pack1: DLEEIMRLENSFKQVLKYVEDAVQ Y218F,Q591F,S668A Disulf1:D217C,S587C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:205 KOs_Cpack1-short_dII- VRVYQKVLIFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG pack1_Disulf2 SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVEWQG dII-dIII-pack1:S367I, IKQKSEVELERLANRSSINLTANSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpack1-short:E686L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L IFNFANSSCVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV dI-dIV-dV-pack1: DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE Y218F,Q591F,S668A DLEEIMRLENSFKQVLKYV Disulf2:D217C,Y589C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:206 KOs_Cpack1-long_dII- VRVYQKVLIFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG pack1_Disulf2 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpack1-long:E686L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L, IFNFANSSCVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV K700A,V702Q DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE dI-dIV-dV-pack1: DLEEIMRLENSFKQVLKYVEDAVQ Y218F,Q591F,S668A Disulf2:D217C,Y589C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:207 KOs_Cpack1-short_dII- VRVYQKVLIFRRSYAYHRTTYLLGSNTTYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRG pack1_N165glyc STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpack1-short:E686L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV dI-dIV-dV-pack1: DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVE Y218F,Q591F,S668A DLEEIMRLENSFKQVLKYV N165glyc:E167T SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:208 KOs_Cpack1-long_dII- VRVYQKVLIFRRSYAYHRTTYLLGSNTTYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRG pack1_N165glyc SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSIVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpackl-long:E686L, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L, IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV K700A,V702Q DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE dI-dIV-dV-pack1: DLEEIMRLENSFKQVLKYVEDAVQ Y218F,Q591F,S668A N165glyc:E167T SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:209 KOs_Cpack1-short_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack2 YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLIFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSIVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack2: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA N220S,V552L IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:210 KOs_Cpack1-long_dI- VRVYQKVLTFRRSYAYHRITYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack2 YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA dI-dIV-dV-pack2: IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV N220S,V552L IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:211 KOs_Cpack1-short_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack3 YSRVIAGIVEVAYHRDSYENKIMQLMPDDESSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack3: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,N220S,V552L, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Q591F,S668A IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:212 KOs_Cpack1-long_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack3 YSRVIAGTVFVAYHRDSYENKTMQLMPDDFSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRENSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA dI-dIV-dV-pack3: IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,N220S,V552L, IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Q591F,S668A DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:213 KOs_Cpack1-short_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGIVEVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRG pack2 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpack1-short:E686L, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV dI-dIV-dV-pack2: DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF N220S,V552L DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:214 KOs_Cpack1-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRG pack2 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVEWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSINLTANSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpack1-long:E686L, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV K700A,V702Q DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE dI-dIV-dV-pack2: DLEEIMRLENSFKQVLKYVEDAVQ N220S,V552L SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:215 KOs_Cpack1-short_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDDFSSTHSTRYVTVKDQWHSRG pack3 SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVICQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA. T374F QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpack1-short:E686L, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV dI-dIV-dV-pack3: DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE Y218F,N220S,V552L, DLEEIMRLENSFKQVLKYV Q591F,S668A SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:216 KOs_Cpack1-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRG pack3 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Cpack1-long:E686L, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Y690F,R693V,V694L, IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV K700A,V702Q DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF dI-dIV-dV-pack3: DLEEIMRLENSFKQVLKYVEDAVQ Y218F,N220S,V552L, Q591F,S668A SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:217 KOs_Cpack1-short_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack3_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATELSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack3: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,N220S,V552L, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Q591F,S668A IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf1:D217C,S587C DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:218 KOs_Cpack1-long_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack3_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRENSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack3: IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,N220S,V552L, IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Q591F,S668A DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF Disulf1:D217C,S587C DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:219 KOs_Cpack1-short_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGIVEVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG pack3_Disulf1 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFELSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Cpack1-short:E686L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Y690F,R693V,V694L QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dII-dIII-pack1:S367I, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV T374F IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV dI-dIV-dV-pack3: DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE Y218F,N220S,V552L, DLEEIMRLENSFKQVLKYV Q591F,S668A Disulf1:D217C,S587C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:220 KOs_Cpack1-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG pack3_Disulf1 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Cpack1-long:E686L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Y690F,R693V,V694L, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA K700A,V702Q IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV dII-dIII-pack1:S367I, IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV T374F DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF dI-dIV-dV-pack3: DLEEIMRLENSFKQVLKYVEDAVQ Y218F,N220S,V552L, Q591F,S668A Disulf1:D217C,S587C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:221 KOs_Cpack1-short_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG pack4_Disulf1 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Cpack1-short:E686L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Y690F,R693V,V694L QLQFTYDTLRGYINPALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dII-dIII-pack1:S367I, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV T374F IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV dI-dIV-dV-pack4: DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF Y218F,N220S,V552L, DLEEIMRLENSFKQVLKYV Q591Y,S668A Disulf1:D217C,S587C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:222 KOs_Cpackl-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG pack4_Disulf1 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Cpack1-long:S686L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Y690F,R693V,V694L, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA K700A,V702Q IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV dII-dIII-pack1:S367I, IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV T374F DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF dI-dIV-dV-pack4: DLEEIMRLENSFKQVLKYVEDAVQ Y218F,N220S,V552L, Q591Y,S668A Disulf1:D217C,S587C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:223 KOs_Cpack1-short_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGIVEVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG pack5_Disulf1 SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRENSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFELSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Cpack1-short:E686L, IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA Y690F,R693V,V694L QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dII-dIII-pack1:S367I, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV T374F IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV dI-dIV-dV-pack5: DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF N220S,V552L,Q591Y DLEEIMRLENSFKQVLKYV Disulf1:D217C,S587C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:224 KOs_Cpack1-long_dII- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG pack5_Disulf1 SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Cpack1-long:E686L, IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA Y690F,R693V,V694L, QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA K700A,V702Q IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV dII-dIII-pack1:S367I, IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV T374F DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF dI-dIV-dV-pack5: DLEEIMRLENSFKQVLKYVEDAVQ N220S,V552L,Q591Y Disulf1:D217C,S587C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:225 KOs_Cpack1-short_dII- VRVYQKVLIFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG pack6_Disulf1 STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVEWQG Cpack1-short:E686L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA Y690F,R693V,V694L QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dII-dIII-pack1:S367I, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV T374F IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV dI-dIV-dV-pack6: DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE N220S,V552L,Q591F DLEEIMRLENSFKQVLKYV Disulf1:D217C,S587C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:226 KOs_Cpack1-long_dII- VRVYQKVLIFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIII-pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG pack6_Disulf1 SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Bg-KOs:I156H,H157R, ASYFGENADKFFIFPNYTIVSDEGRENSALETHRLVAFLERADSVISWD W240N,Y242T,C246S, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM R457S,R460S SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVEWQG Cpack1-long:E686L, IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA Y690F,R693V,V694L, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA K700A,V702Q IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV dII-dIII-pack1:S367I, IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV T374F DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF dI-dIV-dV-pack6: DLEEIMRLENSFKQVLKYVEDAVQ N220S,V552L,Q591F Disulf1:D217C,S587C SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:227 KOs_Cpack1-short_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack4_Disulf1 YSRVIAGTVFVAYHRDSYENKIMQLMPDCFSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack4: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,N220S,V552L, IYNKPIAARFMGDVIGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Q591Y,S668A IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf1:D217C,S587C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:228 KOs_Cpack1-long_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack4_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA dI-dIV-dV-pack4: IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,N220S,V552L, IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Q591Y,S668A DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF Disulf1:D217C,S587C DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:229 KOs_Cpack1-short_dI- VRVYQKVLTFRRSYAYHRITYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack5_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack5: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA N220S,V552L,Q591Y IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Disulf1:D217C,S587C IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCISMKPINEDLDEGIMVVYKRNIVAHTEK NO:230 KosCpack1-long_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack5_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSIVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack5: IYNKPIAARFMGDVIGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV N220S,V552L,Q591Y IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf1:D217C,S587C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:231 KOs_Cpack1-short_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack6_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, SINLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTANSTKSSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack6: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA N220S,V552L,Q591F IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Disulf1:D217C,S587C IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Bg- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:232 KOs_Cpack1-long_dI- VRVYQKVLTFRRSYAYHRTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dIV-dV-pack6_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG Bg-KOs:I156H,H157R, STNLTRETSNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN W240N,Y242T,C246S, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R457S,R460S IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNSTKSSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack6: IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV N220S,V552L,Q591F IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf1:D217C,S587C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_dII-dIII- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:233 pack1 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dII-dIII-pack1:S367I, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG T374F STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSIVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMREENSYKQRVKYV SEQID gB-Towne_Cpack1-short YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:234 Cpack1-short:E686L, VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Y690F,R693V,V694L YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1-long YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:235 Cpack1-long:E686L, VRVYQKVLTERRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Y690F,R693V,V694L, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG K700A,V702Q STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVERDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack2-short YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:236 Cpack2-short:D679N, VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS L680E,E681N,R685Q, YSRVIAGIVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG E686L,Y690F,R693V, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN V694L,K695D ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD IQDEKNVTCQLIFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF NENEIMQLENSFKQVLDYV SEQID gB-Towne_Cpack2-long YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:237 Cpack2-long:D679N, VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS L680E,E681N,R685Q, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG E686L,Y690F,R693V, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN V694L,K695D,K700A, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD V701L,V702Q IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTANRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE NENEIMQLENSFKQVLDYVEDALQ SEQID gB-Towne_Cpack3-short YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:238 Cpack3-short:V677T, VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS F678N,D679N,L680E, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG E681N,E682S,R685Q, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN E686L,N688E,Y690F, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD R693V,V694L,K695D IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTN NENSIMQLFESFKQVLDYV SEQID gB-Towne_Cpack3-long YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:239 Cpack3-long:V677T, VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS F678N,D679N,L680E, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG E681N,E682S,R685Q, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN E686L,N688E,Y690F, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD R693V,V694L,K695D, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM D699K,K700A,V701L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG V702Q IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNIN NENSIMQLFESFKQVLDYVEKALQ SEQID gB-Towne_Cpack4-short YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:240 Cpack4-short:V677T, VRVYQKVLIFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS F678N,D679H,L680E, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG E681N,E682S,M684S, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R685Q,E686L,F687A, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD N688E,Y690F,Q692S, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM R693V,V694L,K695D, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVEWQG Y696R IKQKSLVELERLANRSSLNLTANRIKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTN HENSISQLAESFKSVLDRV SEQID gB-Towne_Cpack4-long YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:241 Cpack4-long:V677T, VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS F678N,D679H,L680E, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVIVKDQWHSRG E681N,E682S,M684S, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN R685Q,E686L,F687A, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD N688E,Y690F,Q692S, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM R693V,V694L,K695D, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y696R,D699K,K700A, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA V701L,V702E QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVERDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTN HENSISQLAESFKSVLDRVEKALE SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:242 short_dII-dIII-pack1 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dII-dIII-pack1:S367I, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG T374F STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack1-short:E686L, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Y690F,R693V,V694L IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:243 long_dII-dIII-pack1 VRVYQKVLIFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dII-dIII-pack1:S367I, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG T374F STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack1-long:S686L, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Y690F,R693V,V694L, IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM K700A,V702Q SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVERDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack2- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:244 short_dII-dIII-pack1 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dII-dIII-pack1:S367I, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG T374F STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN Cpack2-short:D679N, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD L680E,E681N,R685Q, IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM E686L,Y690F,R693V, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG V694L,K695D IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVERDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF NENEIMQLENSFKQVLDYV SEQID gB-Towne_Cpack2- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:245 long_dII-dIII-pack1 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dII-dIII-pack1:S367I, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG T374F STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack2-long:D679N, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD L680E,E681N,R685Q, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM E686L,Y690F,R693V, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG V694L,K695D,K700A, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA V701L,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF NENEIMQLENSFKQVLDYVEDALQ SEQID gB-Towne_Cpack3- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:246 short_dII-dIII-pack1 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dII-dIII-pack1:S367I, YSRVIAGTVFVAYHRDSYENKIMQLMPDDYSNTHSTRYVTVKDQWHSRG T374F STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack3-short:V677T, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD F678N,D679N,L680E, IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM E681N,E682S,R685Q, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG E686L,N688E,Y690F, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA R693V,V694L,K695D QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNTN NENSIMQLFESFKQVLDYV SEQID gB-Towne_Cpack3- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:247 long_dII-dIII-pack1 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dII-dIII-pack1:S367I, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG T374F STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack3-long:V677T, ASYFGENADKFFIFPNYTIVSDEGRENSALETHRIVAFLERADSVISWD F678N,D679N,L680E, IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM E681N,E682S,R685Q, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG E686L,N688E,Y690F IKQKSIVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA R693V,V694L,K695D, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA D699K,K700A,V701L, IYNKPIAARFMGDVLGLASCVTINQTSVKVERDMNVKESPGRCYSRPVV V702Q IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTN NENSIMQLFESFKQVLDYVEKALQ SEQID gB-Towne_Cpack4- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:248 short_dII-dIII-pack1 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dII-dIII-pack1:S367I, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG T374F STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack4-short:V677T, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD F678N,D679H,L680E, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM E681N,E682S,M684S, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG R685Q,E686L,F687A, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA N688E,Y690F,Q692S, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA R693V,V694L,K695D, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y696R IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNIN HENSISQLAESFKSVLDRV SEQID gB-Towne_Cpack4- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:249 long_dII-dIII-pack1 VRVYQKVLTERRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS dII-dIII-pack1:S367I, YSRVIAGIVEVAYHRDSYENKTMQLMPDDYSNTHSTRYVTVKDQWHSRG T374F STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack4-long:V677T, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD F678N,D679H,L680E, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM E681N,E682S,M684S, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG R685Q,E686L,F687A, IKQKSIVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA N688E,Y690F,Q692S, QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA R693V,V694L,K695D, IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y696R,D699K,K700A, IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV V701L,V702E DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNTN HENSISQLAESFKSVLDRVEKALE SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:250 short_dI-dIV-dV-pack1 VRVYQKVLTFRRSYAYINTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Cpack1-short:E686L, YSRVIAGTVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRG Y690F,R693V,V694L STWLYRETCNLNCMVTITTARSKYPYHFFATSIGDVVDISPFYNGTNRN dI-dIV-dV-pack1: ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Y218F,Q591F,S668A IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:251 long_dI-dIV-dV-pack1 VRVYQKVLIFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Cpack1-long:E686L, YSRVIAGTVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRG Y690F,R693V,V694L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN K700A,V702Q ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD dI-dIV-dV-pack1: IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Y218F,Q591F,S668A SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTANRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:252 short_Disulf1 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Cpack1-short:E686L, YSRVIAGIVEVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG Y690F,R693V,V694L STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Disulf1:D217C,S587C ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:253 long_Disulf1 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Cpack1-long:E686L, YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG Y690F,R693V,V694L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN K700A,V702Q ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Disulf1:D217C,S587C IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:254 shortDisulf2 VRVYQKVLIFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Cpack1-short:E686L, YSRVIAGTVFVAYHRDSYENKIMQLMPDCYSNTHSTRYVTVKDQWHSRG Y690F,R693V,V694L STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Disulf2:D217C,Y589C ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSCVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:255 long_Disulf2 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Cpack1-long:E686L, YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG Y690F,R693V,V694L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN K700A,V702Q ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Disulf2:D217C,Y589C IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSCVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:256 short_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG Cpack1-short:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L ASYFGENADKFFIFPNYTIVSDEGRENSALETHRLVAFLERADSVISWD dI-dIV-dV-pack1: IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Y218F,Q591F,S668A SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Disulf1:D217C,S587C IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:257 long_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_Disulf1 YSRVIAGTVFVAYHRDSYENKIMQLMPDCFSNTHSTRYVTVKDQWHSRG Cpack1-long:S686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD K700A,V702Q IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM dI-dIV-dV-pack1: SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y218F,Q591F,S668A IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA Disulf1:D217C,S587C QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF DLEEIMRLFNSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:258 short_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_Disulf2 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG Cpack1-short:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD dI-dIV-dV-pack1: IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Y218F,Q591F,S668A SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Disulf2:D217C,Y589C IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINPALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSCVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:259 long_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_Disulf2 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG Cpack1-long:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L, ASYFGENADKFFIFPNYTIVSDEGRPNSALETARLVAFLERADSVISWD K700A,V702Q IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM dI-dIV-dV-pack1: SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y218F,Q591F,S668A IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA Disulf2:D217C,Y589C QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSCVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:260 short_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTTYVAPPMWEIHHINSHSQCYSS pack1_N165glyc YSRVIAGTVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRG Cpack1-short:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD dI-dIV-dV-pack1: IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Y218F,Q591F,S668A SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG N165glyc:E167T IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:261 long_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTTYVAPPMWEIHHINSHSQCYSS pack1_N165glyc YSRVIAGTVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRG Cpack1-long:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD K700A,V702Q IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM dI-dIV-dV-pack1: SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y218F,Q591F,S668A IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA N165glyc:E167T QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID GB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCISMKPINEDLDEGIMVVYKRNIVAHTFK NO:262 short_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV-pack1 YSRVIAGTVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRG dII-dIII-pack1:S367I, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN T374F ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Cpack1-short:E686L, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Y690F,R693V,V694L SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dI-dIV-dV-pack1: IKQKSLVELERLANRSSLNLTANRTKRSTDGNNATHLSNMESVHNLVYA Y218F,Q591F,S668A QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVEKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:263 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV-pack1 YSRVIAGTVFVAYHRDSYENKTMQLMPDDESNTHSTRYVTVKDQWHSRG dII-dIII-pack1:S367I, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN T374F ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Cpack1-long:E686L, IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFELSKKQEVNM Y690F,R693V,V694L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG K700A,V702Q IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack1: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,Q591F,S668A IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:264 short_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG dII-dIII-pack1:S367I, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN T374F ASYFGENADKFFIFPNYTIVSDFGRENSALETHRLVAFLERADSVISWD Cpack1-short:E686L, IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Y690F,R693V,V694L SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Disulf1:D217C,S587C IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:265 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG dII-dIII-pack1:S367I, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN T374F ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Cpack1-long:E686L, IQDEKNVICQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Y690F,R693V,V694L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVEWQG K700A,V702Q IKQKSLVELERLANRSSLNLTANRIKRSTDGNNATHLSNMESVHNLVYA Disulf1:D217C,S587C QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:266 short_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_Disulf2 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG dII-dIII-pack1:S367I, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN T374F ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Cpack1-short:E686L, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFELSKKQEVNM Y690F,R693V,V694L SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Disulf2:D217C,Y589C IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSCVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:267 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_Disulf2 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSNTHSTRYVTVKDQWHSRG dII-dIII-pack1:S367I, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN T374F ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Cpack1-long:E686L, IQDEKNVICQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Y690F,R693V,V694L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG K700A,V702Q IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA Disulf2:D217C,Y589C QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSCVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:268 short_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKIMQLMPDCFSNTHSTRYVTVKDQWHSRG pack1_Disulf1 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN dII-dIII-pack1:S367I, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD T374F IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSIVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack1: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,Q591F,S668A IYNKPIAARFMGDVLGLASCVTINQTSVKVERDMNVKESPGRCYSRPVV Disulf1:D217C,S587C IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:269 long_dII-dIII- VRVYQKVLIFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKIMQLMPDCFSNTHSTRYVTVKDQWHSRG pack1_Disulf1 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN dII-dIII-pack1:S367I, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD T374F IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVEWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack1: IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,Q591F,S668A IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf1:D217C,S587C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:270 short_dII-dIII- VRVYQKVLTFRRSYAYINTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSNTHSTRYVTVKDQWHSRG pack1_Disulf2 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN dII-dIII-pack1:S367I, ASYFGENADKFFIFPNYTIVSDEGRENSALETHRLVAFLERADSVISWD T374F IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack1: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,Q591F,S668A IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Disulf2:D217C,Y589C IFNFANSSCVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:271 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKIMQLMPDCFSNTHSTRYVTVKDQWHSRG pack1_Disulf2 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN dII-dIII-pack1:S367I, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD T374F IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack1: IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,Q591F,S668A IFNFANSSCVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf2:D217C,Y589C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:272 short_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTTYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRG pack1_N165glyc STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN dII-dIII-pack1:S367I, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD T374F IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Cpack1-short:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack1: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,Q591F,S668A IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV N165glyc:E167T IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:273 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNITYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDDFSNTHSTRYVTVKDQWHSRG pack1_N165glyc STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN dII-dIII-pack1:S367I, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD T374F IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Cpack1-long:E686L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y690F,R693V,V694L, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA K700A,V702Q QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack1: IYNKPIAARFMGDVLGLASCVTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,Q591F,S668A IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV N165glyc:E167T DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF gB-Towne_Cpack1- DLEEIMRLENSFKQVLKYVEDAVQ SEQID short_dI-dIV-dV-pack2 YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:274 Cpack1-short:E686L, VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Y690F,R693V,V694L YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRG dI-dIV-dV-pack2: STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN N220S,V552L ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:275 long_dI-dIV-dV-pack2 VRVYQKVLIFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Cpack1-long:E686L, YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRG Y690F,R693V,V694L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN K700A,V702Q ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD dI-dIV-dV-pack2: IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM N220S,V552L SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:276 short_dI-dIV-dV-pack3 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Cpack1-short:E686L, YSRVIAGTVFVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRG Y690F,R693V,V694L STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN dI-dIV-dV-pack3: ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Y218F,N220S,V552L, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Q591F,S668A SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVFYGQLGEDNEILLGNERTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:277 long_dI-dIV-dV-pack3 VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS Cpack1-long:E686L, YSRVIAGTVFVAYHRDSYENKTMQLMPDDFSSTHSTRYVTVKDQWHSRG Y690F,R693V,V694L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN K700A,V702Q ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD dI-dIV-dV-pack3: IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Y218F,N220S,V552L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Q591F,S668A IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINPALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:278 short_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV-pack2 YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRG dII-dIII-pack1:S367I, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN T374F ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Cpack1-short:E686L, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Y690F,R693V,V694L SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dI-dIV-dV-pack2: IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA N220S,V552L QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:279 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV-pack2 YSRVIAGTVFVAYHRDSYENKTMQLMPDDYSSTHSTRYVTVKDQWHSRG dII-dIII-pack1:S367I, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN T374F ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Cpack1-long:E686L, IQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Y690F,R693V,V694L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG K700A,V702Q IKQKSIVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack2: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA N220S,V552L IYNKPIAARFMGDVIGLASCLTINQTSVKVERDMNVKESPGRCYSRPVV IFNFANSSYVQYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:280 short_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV-pack3 YSRVIAGTVFVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRG dII-dIII-pack1:S367I, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN T374F ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Cpack1-short:E686L, IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM Y690F,R693V,V694L SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dI-dIV-dV-pack3: IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA Y218F,N220S,V552L, QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Q591F,S668A IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:281 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV-pack3 YSRVIAGIVEVAYHRDSYENKTMQLMPDDESSTHSTRYVTVKDQWHSRG dII-dIII-pack1:S367I, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN T374F ASYFGENADKFFIFPNYTIVSDEGRENSALETHRLVAFLERADSVISWD Cpack1-long:E686L, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFELSKKQEVNM Y690F,R693V,V694L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG K700A,V702Q IKQKSIVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA. dI-dIV-dV-pack3: QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,N220S,V552L, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Q591F,S668A IFNFANSSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:282 short_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack3_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG Cpack1-short:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD dI-dIV-dV-pack3: IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Y218F,N220S,V552L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Q591F,S668A IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA Disulf1:D217C,3587C QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:283 long_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack3_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG Cpack1-long:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L, ASYFGENADKFFIFPNYTIVSDEGRENSALETHRLVAFLERADSVISWD K700A,V702Q IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM dI-dIV-dV-pack3: SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y218F,N220S,V552L, IKQKSLVELERLANRSSLNLTANRTKRSTDGNNATHLSNMESVHNLVYA Q591F,S668A QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Disulf1:D217C,S587C IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:284 short_dII-dIII- VRVYQKVLTERRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG pack3_Disulf1 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack1-short:E686L, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Y690F,R693V,V694L IQDEKNVTCQLIFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack3: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,N220S,V552L, IYNKPIAARFMGDVIGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Q591F,S668A IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf1:D217C,S587C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:285 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG pack3_Disulf1 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack1-long:E686L, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Y690F,R693V,V694L, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM K700A,V702Q SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack3: IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,N220S,V552L, IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Q591F,S668A DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE Disulf1:D217C,S587C DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:286 short_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG pack4_Disulf1 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack1-short:E686L, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Y690F,R693V,V694L IQDEKNVICQLTFWEASERTIRSEAEDSYHFISAKMTAFELSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack4: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Y218F,N220S,V552L, IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Q591Y,S668A IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf1:D217C,S587C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:287 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG pack4_Disulf1 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack1-long:E686L, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Y690F,R693V,V694L, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM K700A,V702Q SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack4: IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Y218F,N220S,V552L, IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Q591Y,S668A DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVF Disulf1:D217C,S587C DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCISMKPINEDLDEGIMVVYKRNIVAHTFK NO:288 short_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG pack5_Disulf1 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack1-short:E686L, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Y690F,R693V,V694L IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S367I, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack5: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA N220S,V552L,Q591Y IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Disulf1:D217C,S587C IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:289 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG pack5_Disulf1 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Cpack1-long:E686L, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Y690F,R693V,V694L, IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM K700A,V702Q SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack5: IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV N220S,V552L,Q591Y IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf1:D217C,S587C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:290 short_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG pack6_Disulf1 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN Cpack1-short:E686L, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD Y690F,R693V,V694L IQDEKNVTCQLTFWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM dII-dIII-pack1:S3671, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG T374F IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA dI-dIV-dV-pack6: QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA N220S,V552L,0591F IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV Disulf1:D217C,S587C IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTFK NO:291 long_dII-dIII- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack1_dI-dIV-dV- YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG pack6_Disulf1 STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN Cpack1-long:E686L, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD Y690F,R693V,V694L, IQDEKNVTCQLTEWEASERTIRSEAEDSYHFISAKMTAFFLSKKQEVNM K700A,V702Q SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG dII-dIII-pack1:S367I, IKQKSIVELERLANRSSINLTANRTKRSTDGNNATHLSNMESVHNLVYA T374F QLQFTYDTERGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA dI-dIV-dV-pack6: IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV N220S,V552L,Q591F IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV Disulf1:D217C,S587C DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:292 short_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack4_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCFSSTHSTRYVTVKDQWHSRG Cpack1-short:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGINRN Y690F,R693V,V694L ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD dI-dIV-dV-pack4: IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM Y218F,N220S,V552L, SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Q591Y,S668A IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA Disulf1:D217C,S587C QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDFRVLELYAQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:293 long_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack4_Disulf1 YSRVIAGTVFVAYHRDSYENKIMQLMPDCFSSTHSTRYVTVKDQWHSRG Cpack1-long:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD K700A,V702Q IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM dI-dIV-dV-pack4: SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Y218F,N220S,V552L, IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA Q591Y,S668A QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA Disulf1:D217C,S587C IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYAQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:294 short_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack5_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG Cpack1-short:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD dI-dIV-dV-pack5: IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM N220S,V552L,Q591Y SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Disulf1:D217C,S587C IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:295 long_dI-dIV-dV- VRVYQKVLIFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack5_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG Cpack1-long:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L, ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD K700A,V702Q IQDEKNVTCQLTEWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM dI-dIV-dV-pack5: SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG N220S,V552L,Q591Y IKQKSLVELERLANRSSLNLTANRTKRSTDGNNATHLSNMESVHNLVYA Disulf1:D217C,S587C QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVYYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVE DLEEIMRLENSFKQVLKYVEDAVQ SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:296 short_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack6_Disulf1 YSRVIAGTVFVAYHRDSYENKIMQLMPDCYSSTHSTRYVTVKDQWHSRG Cpack1-short:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L ASYFGENADKFFIFPNYTIVSDFGRPNSALETHRLVAFLERADSVISWD dI-dIV-dV-pack6: IQDEKNVTCQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM N220S,V552L,Q591F SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG Disulf1:D217C,S587C IKQKSLVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA QLQFTYDTLRGYINRALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVERDMNVKESPGRCYSRPVV IFNFANCSYVFYGQLGEDNEILLGNERTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYV SEQID gB-Towne_Cpack1- YRVCSMAQGTDLIRFERNIVCTSMKPINEDLDEGIMVVYKRNIVAHTEK NO:297 long_dI-dIV-dV- VRVYQKVLTFRRSYAYIHTTYLLGSNTEYVAPPMWEIHHINSHSQCYSS pack6_Disulf1 YSRVIAGTVFVAYHRDSYENKTMQLMPDCYSSTHSTRYVTVKDQWHSRG Cpackl-long:E686L, STWLYRETCNLNCMVTITTARSKYPYHFFATSTGDVVDISPFYNGTNRN Y690F,R693V,V694L, ASYFGENADKFFIFPNYTIVSDEGRPNSALETHRLVAFLERADSVISWD K700A,V702Q IQDEKNVICQLTFWEASERTIRSEAEDSYHFSSAKMTATFLSKKQEVNM dI-dIV-dV-pack6: SDSALDCVRDEAINKLQQIFNTSYNQTYEKYGNVSVFETTGGLVVFWQG N220S,V552L,Q591F IKQKSIVELERLANRSSLNLTHNRTKRSTDGNNATHLSNMESVHNLVYA Disulf1:D217C,S587C QLQFTYDTLRGYINPALAQIAEAWCVDQRRTLEVFKELSKINPSAILSA IYNKPIAARFMGDVLGLASCLTINQTSVKVLRDMNVKESPGRCYSRPVV IFNFANCSYVFYGQLGEDNEILLGNHRTEECQLPSLKIFIAGNSAYEYV DYLFKRMIDLSSISTVDSMIALDIDPLENTDERVLELYSQKELRSSNVF DLEEIMRLENSFKQVLKYVEDAVQ

[0140] In some embodiments, the polypeptide comprises, as a C-terminal fusion to the ectodomain, a trimerization domain. In some embodiments, the polypeptide comprises, as a C-terminal fusion to the ectodomain, a nanostructure assembly domain.

[0141] In another aspect, the disclosure provides a fusion protein comprising the polypeptide or antigenic fragment thereof of the disclosure and a multimerization domain.

[0142] In some embodiments, the multimerization domain of the fusion protein of the disclosure comprises a polypeptide with sequence identity of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% to the sequence of a protein listed in Table 12, wherein residues in parentheses are optional

Protein Nanostructures and Assembly Domains

[0143] The gB protein sequences disclosed herein may be displayed on a nanostructure, either as a fusion protein with a component of the nanostructure or by conjugation to the nanostructure. It is contemplated that any of the polypeptides with amino acid substitutions disclosed herein may be used on any of the nanostructures described herein, known in the art, or later developed.

[0144] Various protein nanostructures are known in the art and described, for example in U.S. Pat. Pub. Nos. US2015/0356240A1: US2016/0122392A1. US20180030429A1, US20190341124A1, and US2022/0072120A1, the contents of which are incorporated by reference herein. In some embodiment, the protein nanostructure comprises, as an assembly domain, a variant of KDPG aldolase (Protein Data Bank code 1WA3) engineered to self-assemble into a protein nanostructure.

One-Component Nanostructures

[0145] In its native form. 1WA3 non-covalently assembles to form a trimer via a first interface (the trimer interface). When 20 copies of the trimer (60 monomers) are computationally docked to form a one-component icosahedral protein nanostructure, sets of five monomers of 1WA3 contact one another via a second interface (the pentamer interface). By introducing amino acid substitutions, the pentamer interface may be stabilized such that the protein nanostructure will spontaneously self-assemble, e.g., within the expressing cell or when isolated trimers (or monomers) are mixed under suitable conditions.

[0146] In some embodiments, the pentamer interface comprises 1, 2, 3, 4 or more interface residues, such as amino acid residues in positions 33, 61, 187, and 190 numbered according to SEQ ID NO: 8. In some embodiments, the assembly domain comprises a polypeptide sequence at least 70%, at least 80%, at least 90%, at least 95%, at least 98% identical to SEQ ID NO: 8. In some embodiments, the assembly domain comprises amino acid substitutions at 1, 2, 3, 4 of positions 33, 61, 187, and 190 compared to SEQ ID NO: 8. In some embodiments, a plurality of the amino acid substitutions are substitutions of a polar residue for a non-polar residue (e.g., A, L, I, M, V, F, or W). In some embodiment, some or all of the amino acid substitutions are substitutions of a polar residue for a small, non-polar residue (e.g., A, L, I, M, or V). In some embodiment, the protein nanostructure comprises amino acid substitutions E33L or E33V; K61L or K61M; D187A or D187V; and/or R190A. In some embodiments, the protein nanostructure comprises amino acid substitutions E33L, K61M. D187V, and R190A. In some embodiments, the protein nanostructure comprises amino acid substitutions E33V, K61L, D187A, and R190A. In some embodiments, the assembly domain comprises an amino acid substitution to negate the enzymatic activity of the assembly domain (e.g., K129A). In some embodiments, the assembly domain may comprise further amino acid substitutions (e.g., MI3; E56M or E56K; P186I; E191A; and/or K194A).

[0147] In some embodiments, the assembly domain comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to 13-01 (SEQ ID NO: 8).

[0148] SEQ ID NO: 8 is shown below with illustrative pentamer interface positions in bold:

TABLE-US-00014 (SEQIDNO:8) 1 [00001] 41 [00002] 81 ESGAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKA 121 MKLGHTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVN 161 [00003] 201 RGCTE

[0149] In some embodiments, the assembly domain comprises amino acid substitutions that remove cysteine residues. In some embodiments, the assembly domain comprises C76A and/or C100A substitutions. In some embodiments, the assembly domain comprises C76A, C100A, C165A, and/or C203A substitutions.

[0150] Illustrative assembly domain sequences are provided in Table 11. In each case, the N-terminal MK is optional and not included when calculating sequence identity, but is shown only for numbering purposes, i.e., MK is included in the reference sequence but not necessarily in the assembly domain of the nanostructure.

TABLE-US-00015 TABLE11 SEQ ID Pentamer NO: Sequence Interface 9 (MK)MEELFKKHKIVAVLRANSVEEAKKKALAVFLGGVHLIE none ITFTVPDADTVIKELSFLKEMGAIIGAGTVTSVEQCRKAVES GAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKAMKLG HTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNVCE WFKAGVLAVGVGSALVKGTPVEVAEKAKAFVEKIRGCTE 10 (MK)IEELFKKHKIVAVLRANSVEEAKKKALAVFLGGVHLIE E33L,K61M, ITFTVPDADTVIKELSFLKEMGAIIGAGTVTSVEQCRKAVES D187V,R190A GAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKAMKLG HTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNVCE WFKAGVLAVGVGSALVKGTPVEVAEKAKAFVEKIRGCTE 11 (MK)MEELFKKHKIVAVLRANSVEEAKKKALAVFLGGVHLIE E33L,K61M, ITFTVPDADTVIKELSFLKEMGAIIGAGTVTSVEQARKAVES D187V,R190A GAEFIVSPHLDEEISQFAKEKGVFYMPGVMTPTELVKAMKLG HTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNVCE WFKAGVLAVGVGSALVKGTPVEVAEKAKAFVEKIRGCTE 12 (MK)MEELFKKHKIVAVLRANSVEEAKMKALAVFVGGVHLIE E33V,K61L, ITFTVPDADTVIKELSFLKELGAIIGAGTVTSVEQCRKAVES D187A,R190A GAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKAMKLG HTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNVCE WFKAGVLAVGVGSALVKGTIAEVAAKAAAFVEKIRGCTE 13 (MK)MEELFKKHKIVAVLRANSVEEAKKKALAVFLGGVDLIE E33L,K61M, ITFTVPDADTVIKELSFLKEMGAIIGAGTVTSVEQCRKAVES D187V,R190A GAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKAMKLG HTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNVCE WEKAGVQAVGVGEALNKGTPVEVAEKAKAFVEKIRGCTE 14 (MK)MEELFKKHKIVAVLRANSVEEAKKKALAVFMGGVDLIE E33M,K61L, ITFTVPDADTVIKELSFLKELGAIIGAGTVTSVEQCRKAVES D187A,R190A GAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKAMKLG HTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNVCE WFKAGVQAVGVGEALNKGTPAEVAEKAKAFVEKIRGCTE

Two-Component Nanostructures

[0151] In some embodiments, the assembly domain comprises amino acid substitutions that remove hydrophobic residues and/or add polar residues. Illustrative assembly domains having such substitutions are provided in Int'l Pat. Appl. No. WO2021163481A1, the entire contents of which are incorporated by reference herein. In some embodiments, such amino acid substitutions increase secretion of the assembly domain when the polynucleotides encoding it are expressed in cell.

[0152] SEQ ID NO: 9 is shown below with illustrative positions in bold:

TABLE-US-00016 (SEQIDNO:9) 1 [00004] 41 ITFTVPDADTVIKELSFLKEMGAIIGAGTVTSVEQCRKAV 81 ESGAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKA 121 MKLGHTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVN 161 [00005] 201 RGCTE

[0153] In some embodiments, the assembly domain comprises F32Y; H37D/E/K/N/Q/R; F43Q; F168D/E/K/N/Q/R/S/T/Y; K169D/E/N/Q; L173D/E/N/Q/S; A174S; S179D/E; K183D/E, and/or T185D/E/K/N/Q/S amino acid substitutions.

[0154] In some embodiments, the assembly domain comprises 1, 2, 3, or 4 of H37D, L173Q, S179E, and V182N substitutions. In some embodiments, the assembly domain comprises H37D, L173Q, S179E, and V182N amino acid substitutionsfor example as in SEQ ID NO: 13:

TABLE-US-00017 (SEQIDNO:13) 1 [00006] 41 ITFTVPDADTVIKELSFLKEMGAIIGAGTVTSVEQCRKAV 81 ESGAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKA 121 MKLGHTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVN 161 [00007] 201 RGCTE

[0155] In some embodiments, the mulitimerization domain comprises polypeptides at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of a protein below (Table 12), wherein residues in parentheses are optional. In some embodiments, the mulitimerization domain is covalently linked to a polypeptide or antigenic fragment thereof of CMV gB protein.

TABLE-US-00018 TABLE12 SEQ ID NO: Sequence 15 (MG)KYDGSKLRIGILHARWNAEIILALVLGALKRLQEFGVK I53_dn5A* RENIIIETVPGSFELPYGSKLFVEKQKRLGKPLDAIIPIGVL IKGSTMHFEYICDSTTHQLMKLNFELGIPVIFGVLTCLTDEQ AEARAGLIEGKMHNHGEDWGAAAVEMATKFN 16 (MG)KYDGSKLRIGILHARGNAEIILALVLGALKRLQEFGVK I53_dn5A.1 RENIIIETVPGSFELPYGSKLFVEKQKRLGKPLDAIIPIGVL IRGSTPHFDYIADSTTHQLMKLNFELGIPVIFGVITADTDEQ AEARAGLIEGKMHNHGEDWGAAAVEMATKFN 17 (MG)KYDGSKLRIGILHARGNAEIILELVLGALKRLQEFGVK I53_dn5A.2 RENIIIETVPGSFELPYGSKLFVEKQKRIGKPLDAIIPIGVL IRGSTAHFDYIADSTTHQLMKLNFELGIPVIFGVLTTESDEQ AEERAGTKAGNHGEDWGAAAVEMATKFN 18 (M)EEAELAYLLGELAYKLGEYRIAIRAYRIALKRDPNNAEA I53_dn5B WYNLGNAYYKQGRYREAIEYYQKALELDPNNAEAWYNLGNAY YERGEYEEAIEYYRKALRLDPNNADAMQNLLNAKMREE 19 (M)KMEELFKKHKIVAVLRANSVEEAIEKAVAVFAGGVHLIE I53-50A ITFTVPDADTVIKALSVLKEKGAIIGAGTVTSVEQCRKAVES GAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKAMKLG HTILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNVCE WFKAGVLAVGVGSALVKGTPDEVREKAKAFVEKIRGCTE 20 (M)NQHSHKDYETVRIAVVRARWHAEIVDACVSAFEAAMADI I53-50B GGDRFAVDVFDVPGAYEIPLHARTLAETGRYGAVLGTAFVVN GGIYRHEFVASAVIDGMMNVQLSTGVPVLSAVLTPHRYRDSD AHTLLFLALFAVKGMEAARACVEILAAREKIAA 21 (M)KMEELFKKHKIVAVLRANSVEEAIEKAVAVFAGGVHLIE I53-50A.1 ITFTVPDADTVIKALSVLKEKGAIIGAGTVTSVEQCRKAVES GAEFIVSPHLDEEISQFCKEKGVFYMPGVMTPTELVKAMKLG HDILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNVCE WFKAGVLAVGVGDALVKGDPDEVREKAKKFVEKIRGCTE 22 (M)KMEELFKKHKIVAVIRANSVEEAIEKAVAVFAGGVHLIE I53- ITFTVPDADTVIKALSVLKEKGAIIGAGTVTSVEQCRKAVES 50A.1NegT2 GAEFIVSPHIDEEISQFCKEKGVFYMPGVMTPTELVKAMKLG HDILKLFPGEVVGPEFVEAMKGPFPNVKFVPTGGVDLDDVCE WFDAGVLAVGVGDALVEGDPDEVREDAKEFVEEIRGCTE 23 (M)KMEELFKKHKIVAVLRANSVEEAIEKAVAVFAGGVHLIE I53- ITFTVPDADTVIKALSVLKEKGAIIGAGTVTSVEQCRKAVES 50A.1PosT1 GAEFIVSPHIDEEISQFCKEKGVFYMPGVMTPTELVKAMKLG HDILKLFPGEVVGPQFVKAMKGPFPNVKFVPTGGVNLDNVCK WEKAGVLAVGVGKALVKGKPDEVREKAKKFVKKIRGCTE 24 (M)NQHSHKDHETVRIAVVRARWHAEIVDACVSAFEAAMRDI I53-50B.1 GGDRFAVDVFDVPGAYEIPLHARTLAETGRYGAVLGTAFVVN GGIYRHEFVASAVIDGMMNVQLDTGVPVLSAVLTPHRYRDSD AHTLLFLALFAVKGMEAARACVEILAAREKIAA 25 (M)NQHSHKDHETVRIAVVRARWHAEIVDACVSAFEAAMRDI I53- GGDRFAVDVFDVPGAYEIPLHARTLAETGRYGAVLGTAFVVD 50B.1NegT2 GGIYDHEFVASAVIDGMMNVQLDTGVPVLSAVLTPHEYEDSD ADTLLFLALFAVKGMEAARACVEILAAREKIAA 26 (M)NQHSHKDHETVRIAVVRARWHAEIVDACVSAFEAAMRDI I53- GGDRFAVDVFDVPGAYEIPLHARTLAETGRYGAVLGTAFVVN 50B.4PosT1 GGIYRHEFVASAVINGMMNVQLNTGVPVLSAVLTPHNYDKSK AHTLLFLALFAVKGMEAARACVEILAAREKIAA

[0156] In another aspect, the disclosure provides a nanostructure comprising any polypeptide of the disclosure. In some embodiments, the assembly domain comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to 153-50A (SEQ ID NO: 19), 153-50A.1 (SEQ ID NO: 21), 153-50A.1NegT2 (SEQ ID NO: 22), or 153-50A.1PosT1 (SEQ ID NO: 23). In some embodiments, the assembly domain comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to 153_dn5B (SEQ ID NO: 18). In some embodiments, the nanostructure comprises a second polypeptide. In some embodiments, the second polypeptide comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to 153-50B (SEQ ID NO: 20), 153-50B.1 (SEQ ID NO: 24), 153-50B.1NegT2 (SEQ ID NO: 25), or 153-50B.4PosT1 (SEQ ID NO: 26). In some embodiments, the second polypeptide comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to 153_dn5A* (SEQ ID NO: 15), 153_dn5A.1 (SEQ ID NO: 16), or 153_dn5A.2 (SEQ ID NO: 17).

Ferritin-Based Nanostructures

[0157] In some embodiments, the assembly domain is a ferritin polypeptide. In some embodiments, the assembly domain of a ferritin protein nanostructure comprises a polypeptide sequence at least 70%, at least 80%, at least 90%, at least 95%, at least 98% identical, at least 99% identical, at least 100% identical to any one of the following amino acid sequences:

TABLE-US-00019 (SEQIDNO:27) MLSKDIIKLLNEQVNKEMNSSNLYMSMSSWCYTHSLDGAGLFLFDHAAEE YEHAKKLIIFLNENNVPVQLTSISAPEHKFEGLTQIFQKAYEHEQHISES INNIVDHAIKSKDHATFNFLQWYVAEQHEEEVLFKDILDKVELIGNENHG LYLADQYVKGIAKSRKS (SEQIDNO:28) MLKPEMIEKLNEQMNLELYSSLLYQQMSAWCSYHTFEGAAAFLRRHAQEE MTHMQRLFDYLTDTGNLPRINTVESPFAEYSSLDELFQETYKHEQLITQK INELAHAAMTNQDYPTFNFLQWYVSEQHEEEKLFKSIIDKLSLAGKSGEG LYFIDKELSTLDAQN (SEQIDNO:29) NFHQDCEAGLNRTVNLKFHSSYVYLSMASYFNRDDVALSNFAKFFRERSE EEKEHAEKLIEYQNQRGGRVFLQSVEKPERDDWANGLEALQTALKLQKSV NQALLDLHAVAADKSDPHMTDFLESPYLSESVETIKKLGDHITSLKKLWS SHPGMAEYLFNKHTLG (SEQIDNO:30) QFSKDIEKLLNEQVNKEMQSSNLYMSMSSWCYTHSLDGAGLFLFDHAAEE YEHAKKLIIFLNENNVPVQLTSISAPEHKFEGLTQIFQKAYEHEQHISES INNIVDHAIKSKDHATFNFLQWYVAEQHEEEVLFKDILDKIELIGNENHG LYLADQYVKGIAKSRKSGS (SEQIDNO:31) SGESQVRQNFKPEMEEKLNEQMNLELYSSLLYQQMSAWCSYHTFEGAAAF LRRHAQEEMTHMQRLFDYLTDTGNLPRINTVESPFAEYSSLDELFQETYK HEQLITQKINELAHAAMTNQDYPTFNFLQWYVSEQHEEEKLFKSIIDKLS LAGKSGEGLYFIDKELSTLDGS

[0158] In some embodiments, the assembly domain comprises a polypeptide sequence at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 27, SEQ ID NO: 28, SEQ ID NO: 29, SEQ ID NO: 30, or SEQ ID NO: 31.

Linker

[0159] In some embodiments, the encoded polypeptide includes a linker between regions. A wide variety of polypeptide sequences can be used and are well known in the art. In some embodiments, the linker comprises a Gly-Ser linker (i.e., a linker consisting of glycine and serine residues) of any suitable length. In some embodiments, the Gly-Ser linker is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acids in length. Non-limiting examples of Glys-Ser linkers are presented below:

TABLE-US-00020 (SEQIDNO:298) GS (SEQIDNO:299) GSS (SEQIDNO:300) GSGS (SEQIDNO:301) GSGGSGSGSGGS (SEQIDNO:302) GSGGSGSGSGGS (SEQIDNO:303) GSEKAAKAEEAARK

[0160] In another aspect, the disclosure provides a composition comprising one or more of the polypeptides or fusion proteins of the disclosure linked to a scaffold. The scaffold may be any surface to which the polypeptides or fusion proteins may be bound, including but not limited to a bead, glass, polystyrene, a nanoparticle, a protein scaffold, etc.

[0161] In some embodiments, the scaffold comprises a protein scaffold.

[0162] In some embodiments, the polypeptide is covalently linked to a protein subunit of the protein scaffold to form a fusion protein.

[0163] In some embodiments, the protein subunit of the protein scaffold comprises a polypeptide with sequence identity of at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% to the sequence of a protein listed in Table 12, wherein residues in parentheses are optional.

Polynucleotide

[0164] In another aspect, the disclosure provides a polynucleotide encoding any polypeptide of the disclosure or any nanostructure of the disclosure. The polynucleotide sequence may comprise RNA or DNA. Isolated polynucleotides refers to those that have been removed from their normal surrounding nucleic acid sequences in the genome or in cDNA sequences.

[0165] Such polynucleotide sequences may comprise additional sequences useful for promoting expression and/or purification of the encoded protein, including but not limited to polyA sequences, modified Kozak sequences, and sequences encoding epitope tags, export signals, and secretory signals, nuclear localization signals, and plasma membrane localization signals. It will be apparent to those of skill in the art, based on the teachings herein, what polynucleotide sequences will encode the proteins of the disclosure.

[0166] In some embodiments, the polynucleotide is a messenger RNA (mRNA).

[0167] In another aspect, the disclosure provides a lipid nanoparticle (LNP) comprising an mRNA encoding any polypeptide of the disclosure or any nanostructure of the disclosure.

[0168] In another aspect, the disclosure provides an expression vector comprising any polynucleotide of the disclosure operatively linked to a suitable control sequence. Recombinant expression vector includes vectors that operatively link a nucleic acid coding region or gene to any control sequences capable of effecting expression of the gene product. Control sequences operably linked to the nucleic acid sequences of the disclosure are nucleic acid sequences capable of effecting the expression of the nucleic acid molecules. The control sequences need not be contiguous with the nucleic acid sequences, so long as they function to direct the expression thereof. Thus, for example, intervening untranslated yet transcribed sequences can be present between a promoter sequence and the nucleic acid sequences and the promoter sequence can still be considered operably linked to the coding sequence. Other such control sequences include, but are not limited to, polyadenylation signals, termination signals, and ribosome binding sites. Such expression vectors can be of any type known in the art, including but not limited to plasmid and viral-based expression vectors. The control sequence used to drive expression of the disclosed nucleic acid sequences in a mammalian system may be constitutive (driven by any of a variety of promoters, including but not limited to, CMV, SV40, RSV, actin, EF) or inducible (driven by any of a number of inducible promoters including, but not limited to, tetracycline, ecdysone, steroid responsive). The construction of expression vectors for use in transfecting prokaryotic cells is also well known in the art, and thus can be accomplished via standard techniques. (See, for example, Sambrook, Fritsch, and Maniatis, in: Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory Press, 1989: Gene Transfer and Expression Protocols. pp. 109-128, ed. E. J. Murray, The Humana Press Inc., Clifton, N.J.), and the Ambion 1998 Catalog (Ambion, Austin, TX). The expression vector can be replicable in the host organisms either as an episome or by integration into host chromosomal DNA. Suitable expression vectors and hosts are well known in the art. In some embodiment, the expression vector comprises a plasmid. However, the disclosure is intended to include other expression vectors that serve equivalent functions, such as viral vectors.

[0169] In another aspect, the disclosure provides a host cell comprising any polynucleotide of the disclosure, any expression vector of the disclosure, and/or any polypeptide of the disclosure. The cells can be transiently or stably transfected or transduced. Such transfection or transduction of expression vectors into prokaryotic and eukaryotic cells can be accomplished via any technique known in the art, including but not limited to standard bacterial transformations, calcium phosphate co-precipitation, electroporation, or liposome mediated-, DEAE dextran mediated-, polycationic mediated-, or viral mediated transfection. (See, for example, Molecular Cloning: A Laboratory Manual (Sambrook, et al., 1989, Cold Spring Harbor Laboratory Press; Culture of Animal Cells: A Manual of Basic Technique, 2nd Ed. (R. I. Freshney. 1987. Liss, Inc. New York, NY).

[0170] In some embodiments, the polynucleotide is a messenger RNA (mRNA). Methods of generating a polynucleotide either by chemical synthesis or by in vitro transcription (IVT) (for mRNA) are well known in the art.

[0171] The basic structure of an encoding mRNA can resemble mature eukaryotic mRNA and can include some or all of the following features including (i) a protein-encoding open reading frame (ORF), flanked by (ii) S and 3 untranslated regions (UTRs), and at the end sides (iii) a 7-methyl guanosine 5 cap structure and (iv) a 3 poly(A) tail. The non-coding structural features can be individually optimized to modulate the mRNA stability, translation efficiency, and immunogenicity. By incorporating modified nucleosides, mRNA transcripts referred to as nucleoside-modified mRNA can be produced with reduced immunostimulatory activity, and therefore an improved safety profile can be obtained. In addition, modified nucleosides allow the design of mRNA vaccines with strongly enhanced stability and translation capacity, as they can avoid the direct antiviral pathways that are induced by type IFNs and are programmed to degrade and inhibit invading mRNA. For instance, the replacement of uridine with pseudouridine reduces the activity of 2-5-oligoadenylate synthetase, which regulates the mRNA cleavage by RNase L. In addition, lower activities are measured for protein kinase R, an enzyme that is associated with the inhibition of the mRNA translation process.

[0172] Polynucleotides of the disclosure may include one or more modified (e.g., altered or alternative) nucleobases, nucleosides, nucleotides, or combinations thereof. The polynucleotides can include any useful modification or alteration, such as to the nucleobase, the sugar, or the internucleoside linkage (e.g., to a linking phosphate/to a phosphodiester linkage/to the phosphodiester backbone). In certain some embodiment, alterations (e.g., one or more alterations) are present in each of the nucleobase, the sugar, and the internucleoside linkage. Alterations according to the present disclosure may be alterations of polynucleotides, e.g., the substitution of the 2-OH of the ribofuranosyl of an RNA ring to 2-H, threose nucleic acids (TNAs), glycol nucleic acids (GNAs), peptide nucleic acids (PNAs), locked nucleic acids (LNAs), or hybrids thereof.

[0173] The polynucleotide may include a 5-cap structure. The 5-cap structure of a polynucleotide is involved in nuclear export and increasing polynucleotide stability and binds the mRNA Cap Binding Protein (CBP). A S-UTR may be provided as a flanking region to the mRNA. In some embodiment the polynucleotide sequence is codon-optimized. The polynucleotide may include a polyA sequence or polyadenylation signal, terms used interchangeably.

Pharmaceutical Compositions

[0174] In another aspect, the present disclosure provides pharmaceutical compositions, comprising one or more polypeptides, fusion proteins, protein nanostructures, compositions, nucleic acids, LNPs, expression vectors, and/or host cells of the disclosure and a pharmaceutically acceptable carrier. The pharmaceutical compositions of the disclosure can be used, for example, in the methods of the disclosure described below. The pharmaceutical composition may comprise in addition to the polypeptide of the disclosure (a) a lyoprotectant; (b) a surfactant: (c) a bulking agent: (d) a tonicity adjusting agent: (e) a stabilizer; (f) a preservative and/or (g) a buffer, for example.

[0175] In some embodiments, the buffer is a Tris buffer, a histidine buffer, a phosphate buffer, a citrate buffer or an acetate buffer. The pharmaceutical composition may also include a lyoprotectant, e.g, sucrose, sorbitol or trehalose. In certain some embodiments, the pharmaceutical composition includes a preservative e.g, benzalkonium chloride, benzethonium, chlorohexidine, phenol, m-cresol, benzyl alcohol, methylparaben, propylparaben, chlorobutanol, o-cresol, p-cresol, chlorocresol, phenylmercuric nitrate, thimerosal, benzoic acid, and various mixtures thereof. In other embodiments, the pharmaceutical composition includes a bulking agent, like glycine. In vet other embodiments, the pharmaceutical composition includes a surfactant e.g., polysorbate-20, polysorbate-40, polysorbate-60, polysorbate-65, polysorbate-80 polysorbate-85, poloxamer-188, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan monooleate, sorbitan trilaurate, sorbitan tristearate, sorbitan trioleaste, or a combination thereof. The pharmaceutical composition may also include a tonicity adjusting agent, e.g., a compound that renders the formulation substantially isotonic or isoosmotic with human blood. Exemplary tonicity adjusting agents include sucrose, sorbitol, glycine, methionine, mannitol, dextrose, inositol, sodium chloride, arginine and arginine hydrochloride. In other embodiments, the pharmaceutical composition additionally includes a stabilizer. e.g., a molecule which, when combined with a protein of interest substantially prevents or reduces chemical and/or physical instability of the protein of interest in lyophilized or liquid form. Exemplary stabilizers include sucrose, sorbitol, glycine, inositol, sodium chloride, methionine, arginine, and arginine hydrochloride.

[0176] The polypeptides, fusion proteins, protein nanostructures, compositions, nucleic acids, expression vectors, and/or host cells may be the sole active agent in the pharmaceutical composition, or the composition or vaccine may further comprise one or more other active agents suitable for an intended use.

[0177] The polypeptides, fusion proteins, protein nanostructures, compositions, pharmaceutical compositions, nucleic acids, expression vectors, and/or host cells of the disclosure may be used for any suitable purpose, including but not limited to treat or limit development of CMV infections. For example, the polypeptides, fusion proteins, protein nanostructures, compositions, pharmaceutical compositions, nucleic acids, expression vectors, and/or host cells may be used to elicit an immune response to CMV. One type of immune response is a B-cell response, which results in the production of antibodies against the antigen that elicited the immune response.

[0178] Protective antibodies elicited by methods of this disclosure can protect against viral infections by affecting any step in the life cycle of the virus. For example, protective antibodies may prevent CMV from attaching to a cell, entering a cell, releasing viral ribonucleoproteins into the cytoplasm, forming new viral particles in the infected cell, and/or budding new viral particles from the infected host cell membrane. Antibodies elicited by the methods of this disclosure preferably prevent CMV from attaching to or entering the host cell, prevent fusion of viral membranes with endosomal membranes, or prevent release of newly formed virus from the infected host cell.

[0179] One aspect of this disclosure is a vaccine composition (vaccine) comprising any polypeptide, fusion protein, or composition disclosed herein, to protect subjects against infection by CMV. Vaccines of this disclosure can also contain other components such as adjuvants, buffers and the like. Exemplary adjuvants include aluminum phosphate, benzyalkonium chloride, ubenimex, and QS21: genetic adjuvants such as the IL-2 gene or fragments thereof, the granulocyte macrophage colony-stimulating factor (GM-CSF) gene or fragments thereof, the IL-18 gene or fragments thereof, the chemokine (C-C motif) ligand 21 (CCL21) gene or fragments thereof, the IL-6 gene or fragments thereof, CpG. LPS. TLR agonists, and other immune stimulatory genes: protein adjuvants such IL-2 or fragments thereof, the granulocyte macrophage colony-stimulating factor (GM-CSF) or fragments thereof, IL-18 or fragments thereof, the chemokine (C-C motif) ligand 21 (CCL21) or fragments thereof, IL-6 or fragments thereof, CpG, LPS, TLR agonists and other immune stimulatory cytokines or fragments thereof; lipid adjuvants such as cationic liposomes, N3 (cationic lipid), monophosphoryl lipid A (MPL1); other adjuvants including cholera toxin, enterotoxin. Fms-like tyrosine kinase-3 ligand (Flt-3L), bupivacaine, marcaine, and levamisole.

Methods of Use and Treatment

[0180] This disclosure provides methods of vaccinating a subject against CMV, the method comprising administering polypeptides, fusion proteins, protein nanostructurex, compositions, pharmaceutical compositions, nucleic acids, LNPs, expression vectors, and/or host cells to the subject such that an immune response against CMV is produced in the subject.

[0181] The subject may be any suitable subject, including but not limited to humans and other primates, including non-human primates such as chimpanzees and other apes and monkey species: farm animals such as cattle, sheep, pigs, seals, goats and horses: domestic mammals such as dogs and cats; laboratory animals including rodents such as mice, rats and guinea pigs; birds, including domestic, wild and game birds such as chickens, turkeys and other gallinaceous birds, ducks, geese, and the like. In some embodiments, the subject is a newborn. In some embodiments, the subject is an infant. In some embodiments, the subject is pregnant. In some embodiments, the subject is immunocompromised or is at risk of becoming immunocompromised such as a candidate for organ, stem cell or bone marrow transplant.

[0182] In the vaccination methods of this disclosure, the subject being vaccinated may have been exposed to CMV. As used herein, the term exposed indicate the subject has come in contact with a person or animal that is known to be infected with CMV. Vaccines of this disclosure may be administered by any suitable technique, by means including, but not limited to, traditional syringes, needleless injection devices, or microprojectile bombardment gene guns. Suitable routes of administration include, but are not limited to, parenteral delivery, such as intramuscular, intradermal, subcutaneous, or intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, or intraocular injection.

[0183] A composition provided herein may be co-administered with other treatments, such as other vaccines. In some embodiments, a subject treated in accordance with a method provided herein may also be administered one or more seasonal or pandemic vaccines such as an influenza vaccine or a SARS-Cov2 vaccine. In some embodiments, a subject treated in accordance with the methods provided herein may also be administered a pneumococcal. Recombinant Zoster (Shingles), or Tdap vaccine. One, two, or more vaccines may be co-administered with an immunogenic composition provided herein. Co-administration includes both concurrently as well as subsequent administration. For example, the one, two, or more vaccines and an immunogenic composition provided herein may be administered on the same day. In some embodiment, the one, two, or more vaccines and an immunogenic composition provided herein are administered within 1 hour, within 2 hours, within 3 hours, within 4 hours, within 5 hours, within 6 hours, within 8 hours, withing 10 hours, or within 12 hours.

[0184] In another aspect, provide herein is a method of treating a subject suffering from viral infection. As used herein, treat or treating includes, but is not limited to accomplishing one or more of the following: (a) reducing viral titer in the subject: (b) limiting any increase of viral titer in the subject: (c) reducing the severity of viral infection symptoms; (d) limiting or preventing development of symptoms after viral infection: (e) inhibiting worsening of symptoms of viral infection: (f) limiting or preventing recurrence of symptoms of viral infection in subjects that were previously symptomatic for viral infection; and/or (g) increasing survival. In some embodiments, a method of vaccinating decreases the subject's risk of becoming infected with a virus. In some embodiments, a method of vaccinating limits the development of a viral infection. In some embodiments, a method of vaccinating decreases the severity of the symptoms of a viral infection.

[0185] In some embodiment, the methods provided herein may be used to prevent a viral infection or illness (e.g., pneumonia or acute respiratory disease) in a subject. As used herein, prevent or preventing includes, but is not limited to accomplishing one or more of the following: (a) generating an immune response (antibody and/or cell-based, e.g., CD4 T cells, memory B cells, and/or CD8 T cells) to a virus in the subject expected to confer protection against infection caused by or associated with Cytomegalovirus in the subject: (b) generating neutralizing antibodies against Cytomegalovirus in the subject expected to reduce the severity of one or more Cytomegalovirus-caused symptoms in the subject; (c) preventing

[0186] Cytomegalovirus infection in a subject, detected as an increase in the titer of the virus of the subject or by an increase in one or more symptoms of viral infection: (d) reducing the risk of Cytomegalovirus infection within a population of subjects; or (e) causing a seroresponse (or seroconversion) of a subject, such as generating neutralizing antibodies against Cytomegalovirus at least 4-fold higher than a baseline antibody level in the subject. Prevention may be assessed by comparing immune responses, especially correlates of protection, in subjects administered a vaccine to the same subjects before administration (termed baseline), subjects administered a placebo, or subjects administered a comparator vaccine.

[0187] As used herein, limiting the development a viral infection, refers to accomplishing one or more of the following: (a) generating an immune response (antibody and/or cell-based, e.g., CD4 T cells, memory B cells, and/or CD8 T cells) to Cytomegalovirus in the subject expected to limit an increase in viral titer or symptoms in the subject: (b) generating neutralizing antibodies against Cytomegalovirus in the subject at a level expected to limit an increase in viral titer or symptoms in the subject: (c) causing reduced viral titers in the subject after exposure to Cytomegalovirus compared to subjects not administered the protein complex; and (d) caused reduced incidence or severity of symptoms after Cytomegalovirus infection. Illustrative symptoms of Cytomegalovirus infection include, but are not limited to, fever, fatigue, swollen glands, sore throat and muscle ache.

[0188] Further, the methods provided herein may be used to prevent or limit development of infection with an original strain of Cytomegalovirus and/or infection with a variant strain of Cytomegalovirus.

[0189] Clinical efficacy of a Cytomegalovirus vaccine can be assessed by various means known in the art, including but not limited to placebo-controlled clinical efficacy studies to measure viral load or symptoms of viral disease in vaccinated versus control subjects. Correlates of protection may also be defined, such as neutralizing antibody titers (typically expressed as geometric mean titers), fold increases above baselines (typically expressed as geometric fold rise), and seroresponse rate (a percentage of subjects that achieve a fold rise in neutralizing antibody titers above a predetermined threshold). Guidance on direct and surrogate measures of clinical efficacy for respiratory diseases are available, for example, in Guidance for Industry: Clinical Data Needed to Support the Licensure of Seasonal Inactivated Influenza Vaccines. U.S. Food & Drug Administration (May 2007) and Respiratory Syncytial Virus Infection: Developing Antiviral Drugs for Prophylaxis and Treatment Guidance for Industry. U.S. Food & Drug Administration (October 2017).

[0190] In some embodiments, the methods described herein generate an immune response in a subject in the subject not known to be infected with a virus, wherein the immune response serves to limit development of infection and symptoms of a viral infection. In some embodiments, the immune response comprises generation of neutralizing antibodies and/or cell-based responses against a virus. In some embodiments, the immune response comprises generation of gB protein-specific or other trimer-specific responses with a mean geometric titer of at least 110.sup.3, at least 110.sup.4, at least 110.sup.5, at least 110.sup.6, at least 110.sup.7, at least 110.sup.8, or at least 110.sup.9. In a further embodiment, the immune response comprises generation of antibodies against multiple antigenic epitopes on the gB protein-specific or other viral protein trimer.

[0191] In another aspect, the disclosure provides a method of generating an immune response to CMV gB protein in a subject in need thereof, comprising administering to the subject, an effective amount of the pharmaceutical composition of the disclosure.

[0192] In one aspect, the methods provided herein may result in an increase in antibody titers in a subject, e.g., in an increase in virus-specific neutralizing antibodies or virus-specific binding antibodies. Antibody titers may be determined using any suitable assays known in the art or described herein including, without limitation, binding enzyme-linked immunosorbent assays (ELISA), enzyme-linked immune absorbent spot (ELISpot), competition ELISAs, immunoprecipitation, immunoblotting, and agglutination assays.

[0193] In some embodiments, a neutralization or microneutralization (MN) assay may be used to measure increases in neutralizing antibodies in a subject after administration of a protein complex as described herein. Microneutralization refers to a neutralizing assay performed in a miniaturized format, such as a 96-well plate. A (micro) neutralization assay is used to test for the inhibition of a virus by antibodies (e.g., purified antibodies, serum, or plasma). The assay measures the level of antibodies present in a sample that are able to neutralize a virus in vitro. Generally, microneutralization assays for clinical samples are performed with a serial dilution of serum mixed with a fixed concentration of virus. Methods for performing (micro) neutralization assays are well known. Illustrative microneutralization assays are described, see, e.g., van Baleen et al. Vaccine 35 (2017) 46-52.

[0194] If neutralizing antibodies specific for Cytomegalovirus are present in a sample, the virus will be neutralized, and infection of cells (e.g., HEp-2 cells) is inhibited. Immunofluorescence levels indicating viral infection may be analyzed using, e.g., a CTL ImmunoSpot UV analyzer, equipped with BioSpot analysis software for automated counting of infected cells. Results are generally reported in international units per milliliter (IU/mL).

[0195] By baseline is meant a measurement of antibodies prior to administration of the first dose of an immunogenic composition provided herein.

[0196] In another aspect, the methods provided herein may result in an increase in immune cells in a subject (e.g., an increase in virus-specific memory B cells and/or virus-specific T cells). The number of immune cells in a subject may be determined using any suitable assay known in the art or described herein, including, without limitation, FACS and flow cytometry.

[0197] In some embodiments, the composition is administered via any suitable route, including intranasally, sublingually, orally, parentally, by inhalation spray, rectally, or topically in dosage unit formulations containing conventional pharmaceutically acceptable carriers, adjuvants, and vehicles. The term parenteral as used herein includes, subcutaneous, intravenous, intra-arterial, intramuscular, intrasternal, intratendinous, intraspinal, intracranial, intrathoracic, infusion techniques or intraperitoneally.

[0198] In another aspect, the disclosure provides a method of treating or preventing CMV infection in a subject in need thereof, comprising administering to the subject, an effective amount of the pharmaceutical composition of the disclosure.

[0199] In another aspect, the disclosure provides a vaccine comprising one or more of the polypeptides, fusion protein, protein nanostrucure, composition, nucleic acid, LNP expression vector, and/or the host cell of the disclosure and a pharmaceutically acceptable carrier.

[0200] In some embodiments, the vaccine of the disclosure further comprises any other component as appropriate for an intended use, including but not limited to any other CMV antigens, including but not limited to one or more of CMV proteins gH, gL, UL128, UL130 and UL131, or antigenic portions thereof; or a pentamer complex of CMV proteins gH, gL, UL128, UL130 and UL131 or antigenic portions thereof.

[0201] In another aspect, the disclosure provides a method for treating or limiting development of an CMV infection, comprising administering to a subject in need thereof an amount effective to treat or limit development of the CMV infection of a polypeptide, fusion protein, composition, vaccine, nucleic acid, expression vector, host cell, pharmaceutical composition, and/or vaccine of the disclosure.

Kits

[0202] The disclosure further provides kits, which may be used to prepare and administer compositions of the disclosure. In some embodiments, a kit provided herein comprises a composition as disclosed herein, and instructions for use in a method of the disclosure. In some embodiments, a kit comprises one or more unit doses as disclosed herein, and instructions for use in a method of the disclosure. In some embodiments, the kit comprises a vial comprising a single dose of a pharmaceutical composition provided herein. In some embodiments, a kit comprises a vial comprising multiple doses provided herein. In some embodiments, a kit further comprises instructions for use of the pharmaceutical composition. In some embodiments, a kit further comprises a diluent for preparing dilutions of the pharmaceutical composition prior to administration. In some embodiments, the pharmaceutical composition comprises an adjuvant. In some embodiments, a kit comprises a pharmaceutical composition and an adjuvant which may be mixed prior to administration. In another aspect, the disclosure provides a composition, method, or use as described herein.

EXAMPLES

[0203] Provided are sequences of recombinant CMV gB proteins in which, while not wishing to be held by theory, mutations have been added to disrupt the postfusion state and/or stabilize the prefusion state of the gB protein. Regions of gB (FIG. 1A) were identified with sub-optimal local stability for the prefusion state, and specific mutations which are intended to both improve these structural features and/or destabilize the postfusion state are provided. For example, insertion of a glycosylation site at a position exposed in the postfusion conformation is shown in FIG. 2B (compared to prefusion. FIG. 2A). Designed gB antigens have been characterized by size exclusion chromatography (SEC) as trimeric but show distinctly different biophysical profiles as assessed by nano differential scanning fluorimetry (nanoDSF) and dynamic light scattering (DLS). Further negative stain electron microscopy (NS-EM) confirms that designed CMV antigens less often favor the postfusion state, while instead showing minor conformations that better resemble the prefusion state. Together, the mutations provided herein are potentially useful for diverse forms of vaccines containing recombinant gB antigens, including protein-based, nucleic acid-based or vectored vaccines.

Example 1: Materials and Methods

[0204] Expression and protein purification: Genes encoding mutant CMV gB antigens (corresponding to residues 20-698, 20-702, or 20-704 of SEQ ID NO: 1) were cloned in the pCMV/R vector using the XbaI and AvrII restriction sites. In addition to various designed mutations, all sequences also contained the mutations I156H, H157R, W240N. Y242T, C246S, R457S, R460S to make fusion loops more polar, remove an unpaired cysteine, and remove a poly-basic cleavage site. All sequences were preceded by a MGILPSPGMPALLSLVSLLSVLLMGCVAETGT (SEQ ID NO: 304) secretion signal and C-terminally tagged with either a GGSHHHHHHHH (SEQ ID NO: 305) sequence to allow for purification, a GSGYIPEAPRDGQAYVRKDGEWVLLSTFLGGHHHHHHHH (SEQ ID NO: 306) histidine-tagged T4 fibritin foldon sequence to allow for both improved trimerization and purification, or a GSRMKQIEDKIEEILSKIYHIENEIARIKKLIGERGGHHHHHHHH (SEQ ID NO: 307) histidine-tagged GCN4 sequence to allow for both improved trimerization and purification.

[0205] All proteins were produced in Expi293F cells grown in suspension using Expi293F expression medium (Life Technologies) at 37 C., 70% humidity. 8% CO.sub.2 rotating at 150 rpm. Cell cultures were transfected using PEI-MAX (Polyscience) with cells grown to a density of 3.0 million cells per mL and cultivated for 3 days. Supernatants were clarified by centrifugations at 4000 ref, addition of PDADMAC to a final concentration of 0.0375% (Sigma Aldrich) and a second spin at 4000 ref.

[0206] His-tagged proteins were purified from clarified supernatants via a batch bind method where each clarified supernatant was supplemented with 1 M Tris-HCl pH 8.0 to a final concentration of 45 mM and 5 M NaCl to a final concentration of 310 mM. Talon cobalt affinity resin (Takara) was added to the treated supernatants and allowed to incubate for 15 minutes with gentle shaking. Resin was collected using vacuum filtration with a 0.2 m filter and transferred to a gravity column. The resin was washed with 20 mM Tris pH 8.0, 300 mM NaCl, and the protein was eluted with 3 column volumes of 20 mM Tris pH 8.0, 300 mM NaCl, 300 mM imidazole. Protein purity after affinity purification was assessed by SDS-PAGE (both reducing and non-reducing). Eluates were concentrated and applied to a Superose 6 Increase 10/300 column (Cytiva) pre-equilibrated with 25 mM tris pH 8.0, 150 mM NaCl, 5% glycerol for preparative size exclusion chromatography (SEC). Peaks corresponding to the trimeric species were identified based on elution volume. Purified protein was stored at 4 C.

[0207] Thermal melts: All proteins were diluted to 1 mg/mL in 25 mM tris pH 8.0, 150 mM NaCl, 5% glycerol prior to thermal measurements. Measurements of melting temperatures were determined from thermal denaturation melt curves using an UNcle (UNchained Labs) based on the barycentric mean (BCM) of intrinsic tryptophan fluorescence, with data collected from 20-95 C. using a thermal ramp of 1 C. per minute in a background of 25 mM Tris pH 8.0, 150 mM NaCl and 5% glycerol. Melting temperatures were calculated using UNcle analysis software.

[0208] Negative stain electron microscopy: Protein was adsorbed to glow-discharged carbon-coated copper grids for 1 min prior to a 3 wash with water and 2% uranyl formate staining Micrographs were recorded using the Leginon software on a 100 kV FEI Tecnai G2 Spirit with a Gatan Ultrascan 4000 4k4k CCD camera at 67,000 nominal magnification. The defocus ranged from 1.0 to 2.0 mm and the pixel size was 1.6 . Particles were picked automatically in a reference free manner using DogPicker. Contrast transfer function (CTF) estimation was performed using GCTF. Class averages were generated using Relion2.1.

Example 2: Computational Design of gB Protein

[0209] Rationale structure-based design using the crystal structure of CMV gB protein (PDB 7KDP) was used to identify amino-acid substitutions that would stabilize the prefusion conformation or destabilize the postfusion conformation of gB protein. Briefly, the prefusion (PDB 7KDP) and postfusion (PDB 7KDD) structures of CMV gB were analyzed using PyMol software to identify regions with different structural conformations in either state. For C terminus and domain-domain groups (Table 13), multiple residues in these regions were selected for design using computational protocols from Rosetta software (Leman et al. Nat. Methods 17 (7): 665-680 (2020)), in which selected residues were allowed to be simultaneously mutated by Rosetta to improve noncovalent interactions within the prefusion structure. Output sequences from Rosetta with different types and/or numbers of mutations were scored using Rosetta in both prefusion and postfusion structures to predict energetic effects towards both states (Table 13. energetic difference), and sets of mutations that were predicted to stabilize the prefusion state and/or destabilize the postfusion state were selected for experimental analysis. For the disulfide group (Table 13), pairs of residues were selected that are distant in the postfusion structure, but in close enough proximity in the prefusion structure to form a disulfide bond if both mutated to cysteine. For the glycosylation group (Table 13), the E167T mutation was made to glycosylate N165. N165 is strongly buried in the postfusion structure and not capable of accommodating an added glycan without introducing steric clashes, but solvent-exposed in the prefusion structure to allow ample space for an added glycan.

[0210] Table 13 shows predicted computationally predicted stabilization of CMV gB protein. Representative wild-type sequence (SEQ ID NO: 1-3) was modified to include amino acid substitutions at I156H, H157R. W240N. Y242T, C246S, R457S, R460S. This representative sequence was further modified as in Table 13.

TABLE-US-00021 TABLE 13 Summary of CMV mutation data Energenic difference (preTrimer (mutPre preMonomer) wtPre) preTrimer (postTrimer (mutPost Group Mutant Mutations preMonomer postMonomer) wtPost) C Cpack1- E686L, Y690F, 156 REU postfusion 32 REU terminus short R693V, V694L 14 REU Cpack1- E686L, Y690F, 212 REU prefusion 406 REU long R693V, V694L, 41 REU K700A, V702Q Cpack2- D679N, L680E, 166 REU postfusion 24 REU short E681N, R685Q, 6 REU E686L, Y690F, R693V, V694L, K695D Cpack2- D679N, L680E, 236 REU prefusion 396 REU long: E681N, R685Q, 64 REU E686L, Y690F, R693V, V694L, K695D, K700A, V701L, V702Q Cpack3- V677T, F678N, 166 REU prefusion 168 REU short D679N, L680E, 32 REU E681N, E682S, R685Q, E686L, N688E, Y690F, R693V, V694L, K695D Cpack3- V677T, F678N, 179 REU prefusion 573 REU long D679N, L680E, 113 REU E681N, E682S, R685Q, E686L, N688E, Y690F, R693V, V694L, K695D, D699K, K700A, V701L, V702Q Cpack4- V677T, F678N, 243 REU prefusion 325 REU short D679H, L680E, 104 REU E681N, E682S, M684S, R685Q, E686L, F687A, N688E, Y690F, Q692S, R693V, V694L, K695D, Y696R Cpack4- V677T, F678N, 261 REU prefusion 562 REU long D679H, L680E, 122 REU E681N, E682S, M684S, R685Q, E686L, F687A, N688E, Y690F, Q692S, R693V, V694L, K695D, Y696R, D699K, K700A, V701L, V702E Domain- dII-dIII- S367I, T374F No effect on trimer stability in either 67 REU domain pack1 prefusion or postfusion conformation. dI-dIV- Y218F, Q591F, Slightly postfusion 62 REU dV-pack1 S668A stabilizes the 9 REU trimer 14 REU dI-dIV- N220S, V552L 50 REU prefusion 158 REU dV-pack2 -38 REU dI-dIV- Y218F, N220S, 62 REU prefusion 177 REU dV-pack3 V552L, Q591F, -33 REU S668A dI-dIV- Y218F, N220S, 61 REU prefusion 189 REU dV-pack4 V552L, Q591Y, -32 REU S668A dI-dIV- N220S, V552L, 45 REU prefusion 252 REU dV-pack5 Q591Y -33 REU dI-dIV- N220S, V552L, 48 REU prefusion 151 REU dV-pack6 Q591F -36 REU C-beta distance C-beta distance in in prefusion postfusion Disulfide Disulf1 D217C, S587C 3.9 Angstrom 134.6 Angstrom Disulf2 D217C, Y589C 4.4 Angstrom 138.4 Angstrom Glycosylation N165glyc E167T N/A

[0211] The first value of Table 13 is the energetic difference between the prefusion trimer and the prefusion monomer (pre Trimer-preMonomer). The second value of Table 13 is the difference between the above and the equivalent difference for the postfusion state ((pre Trimer-preMonomer)(post Trimer-postMonomer)). The final value of the Table 13 is the difference is energy change between pre- and postfusion states by adding the mutations ((mutPre-wtPre)(mutPost-wtPost)). REU means Rosetta Energy Units. Methods for calculating these parameters are provided in Park et al. J. Chem. Theory & Computation 12 (12) 6201-12 (2016). Monomer calculations come from a model where each chain in the trimer is translated 1000 Angstrom apart to simulate a dissociated state.

Example 3: Results

[0212] Referring to FIG. 3, recombinant gB designs tested all included at least the soluble ectodomain of gB from the Towne strain from residues 20-698, with some designs containing additional residues up to residue 704. All designs tested featured an exogenous N-terminal signal peptide and a C-terminal octa-histidine tag, with some designs also featuring an exogenous trimerization domain between the C-terminus of the gB ectodomain and the octa-histidine tag. All designs tested also featured polar mutations in the fusion loop to assist protein solubility (I156H. H157R, W240N, Y242T), a mutation to an unpaired cysteine (C246S) and mutations to a polybasic cleavage site (R457S, R460S). Collectively, these are termed Bg-KO. Without being bound by theory, these mutations should impact the expression level but not the prefusion:postfusion ratio. Therefore, it is expected that constructs lacking these amino acid substitutions would still have the desired prefusion preference induced by the designed mutations described in Example 2 and tested here. In other words, the Bg-KO substitutions are an optional feature of the constructs.

[0213] FIG. 4 shows size exclusion chromatography (SEC) (Superose 6 Increase 10/300 GL, Cytiva) of multiple gB antigens featuring designed mutations. The designed constructs had homogenous elution profiles consistent with trimeric gB antigens. A comparable postfusion gB antigen lacking designed mutations is shown as a reference. The dimensions of the postfusion structure of gB (PDB 7KDD) notably feature a great length (17 nm) and a relatively small width (7 nm). Because SEC elution volume is influenced by a molecule's radius of gyration and hydrodynamic diameter, prefusion gB should eluate later than postfusion gB. The five different designs with the Cpack1-long mutations alone or with additional mutations show later elution volumes, which is consistent with the samples containing an increase ratio of prefusion to postfusion gB.

[0214] FIG. 5 shows dynamic light scattering (DLS) of multiple gB antigens featuring designed mutations, showing distinctly different hydrodynamic diameters and polydispersities, which is indicative of conformational changes. A comparable gB antigen lacking designed mutations is shown as a reference. Prefusion gB should have a larger hydrodynamic diameter than postfusion gB. Cpack1-long, in combination with Disulf1, dII-dIII pack 1 and GCN4, or Disulf 1 and di-dIV-dV-pack 1, show larger DLS diameter than the postfusion control.

[0215] FIG. 6 shows nano differential scanning fluorimetry (nanoDSF) of multiple gB antigens featuring designed mutations, with nanoDSF measured using intrinsic tryptophan fluorescence. A comparable gB antigen lacking designed mutations is shown as a reference. Prefusion gB should be less thermally stable than postfusion gB, due to the low energy of the postfusion state needed for membrane fusion. Compared to the postfusion control, which showed two melting transitions, addition of designed mutations led to different thermal profiles in which one or more melting temperatures was lowered.

[0216] FIG. 7 shows negative stain electron micrographs (NS-EM) micrographs of multiple gB antigens featuring designed mutations. A comparable gB antigen lacking designed mutations is shown as a reference, which more monodispersely features an elongated profile consistent with the postfusion state. All constructs with designed mutations showed less postfusion character compared to the postfusion control. The combination of Cpack1-long and other design features led to greater prevalence of conformations that are not similar to the postfusion structure. Specifically, particles with more spherical character were observed with the combination of Cpack1-long, Disulf1 and dI-dIV-dV-pack 1 mutations, which is more similar to the prefusion structure.

[0217] FIG. 8 shows NS-EM 2D class averages of one gB antigen featuring designed mutations and a comparable gB antigen lacking designed mutations.

[0218] The foregoing description of the embodiments of the invention has been presented for the purpose of illustration; it is not intended to be exhaustive or to limit the invention to the precise forms disclosed. Persons skilled in the relevant art can appreciate that many modifications and variations are possible in light of the above disclosure. Accordingly, the scope of the invention is limited only by the following claims.