Glycosylated engineered interferon beta proteins

Abstract

Disclosed herein are engineered, non-naturally occurring protein variants with de novo glycosylation sites having wildtype-like biological activity of wildtype interferon beta. Also disclosed herein are vectors encoding the non-naturally occurring protein variants described herein. Also disclosed herein are pharmaceutical compositions comprising the non-naturally occurring protein variants described herein or a vector encoding the non-naturally occurring protein variants described herein. Also disclosed herein are methods of treating a disease by administering to a subject the non-naturally occurring protein variants described herein, a vector encoding the non-naturally occurring protein variants described herein, or a pharmaceutical composition comprising the non-naturally occurring protein variants described herein or a vector described herein.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority under 35 U.S.C. § 119 from U.S. Provisional Application No. 63 / 566,081 filed on March 15, 2024, the disclosure of which is incorporated herein by reference in their entirety. STATEMENT AS TO FEDERALLY SPONSORED RESEARCH

[0002] This invention was made with government support under Contract number 1R43HG012537-01A1 awarded by The National Institutes of Health. The government has certain rights in the invention. SUMMARY

[0003] Provided herein are engineered interferon beta (IFN-beta) proteins or a functional fragments thereof, comprising a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1. In some embodiments, the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208. In some embodiments, the modification comprises an amino acid substitution, a deletion, or an insertion in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modification is an amino acid substitution, and the amino acid substitution comprises at least one amino acid substitution in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modification is a deletion of one or more amino acids in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modification comprises an insertion of one or more amino acids in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the at least one amino acid substitution results in an increase in a biological activity of the engineered IFN-beta protein as compared to a biological activity of the wildtype protein. In some embodiments, the at least one amino acid substitution results in a decrease in a biological activity of the engineered IFN-beta protein as compared to a biological activity of the wildtype protein. In some embodiments, the increase in the biological activity of biological activity comprises an increase in signaling, an increase in protein expression of one or more biological targets, an increase in mRNA expression, or any combination thereof, as measured by an in vitro assay. In some embodiments, the increase in signaling comprises an increase in IFNAR1-mediated signaling, as measured by an in vitro assay. In some embodiments, increase in signaling comprises an increase in IFNAR2-mediated signaling, as measured by an in vitro assay. In some embodiments, wherein the modification comprises: (a) an amino acid substitution that results in a conversion of a motif in the amino acid sequence of SEQ ID NO: 1 that is not a glycosylation motif into a glycosylation motif, thereby introducing a glycosylation motif into the engineered protein; (b) an amino acid substitution that results in a conversion of a glycosylation motif in the amino acid sequence of SEQ ID NO: 1 into a motif that is not a glycosylation motif, thereby eliminating a glycosylation motif from the engineered protein; or (c) a combination of (a) and (b). In some embodiments, the at least one amino acid substitution results in the conversion of 1, 2, 3, 4, or more motif(s) in the amino acid sequence of SEQ ID NO: 1 that are not glycosylation motifs into 1, 2, 3, 4, or more motif(s) that are glycosylation motif(s), thereby introducing 1, 2, 3. 4, or more glycosylation motif(s) into the engineered protein. In some embodiments, the at least one amino acid substitution results in the conversion of 1,2, 3, 4, or more glycosylation motif(s) in the amino acid sequence of SEQ ID NO: 1 into 1, 2, 3, 4, or more motif(s) that are not glycosylation motifs, thereby eliminating 1, 2, 3, 4, or more glycosylation motif(s) into the engineered protein. In some embodiments, the glycosylation motif has a consensus sequence of N-X-S / T, wherein N is an Asparagine that is N-glycosylated at the amide bond in the side chain of the Asparagine, X is any amino acid except for proline, S is a serine, and T is a threonine. In some embodiments, the modification comprises an amino acid substitution that results in a conversion of a motif in the amino acid sequence of SEQ ID NO: 1 that is not a site for a post-translational modification into a post-translational modification consensus sequence motif, thereby introducing a site for a post translational modification into the engineered protein. In some embodiments, the post-translational modification consensus sequence motif is selected from the amino acid sequences in TABLE 5. In some embodiments, the amino acid substitution results in the addition of a post-translational modification on the engineered protein at the site of the post-translational modification consensus sequence motif that is not present in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the post-translational modification is selected from the group consisting of: glycosylation, phosphorylation, lipidation, glycation, proline isomerization, methylation, acetylation, hydroxylation sites, and proteolysis. In some embodiments, the engineered protein is glycosylated. In some embodiments, the engineered protein is aglycosylated. In some embodiments, the engineered protein binds to an interferon-a / p receptor-1(IFNARI) or an interferon-a / p receptor-2 (IFNAR2) heterodimer subunits at a level greater than or equal to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. as measured by enzyme-linked immunosorbent assay (ELISA), or by measuring signaling activity of an Interferon Stimulated Response Element (ISRE)-Green fluorescent protein (GFP) reporter in a live human cell in vitro assay. In some embodiments, the engineered protein binds to and activates an IFNAR1 or an IFNAR2 heterodimer subunits at a level greater than or equal to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by ELISA, or by measuring signaling activity of an ISRE-GFP reporter in a live human cell in vitro assay. In some embodiments, the engineered protein binds to and inhibits an activity of an IFNAR1 or an IFNAR2 heterodimer subunits at a level greater than or equal to the wildlype IFN-beta protein as measured by ELISA, or by measuring signaling activity of an ISRE-GFP reporter in a live human cell in vitro assay. In some embodiments, the engineered protein is an agonist of IFNAR1 or IFNAR2 subunits. In some embodiments, the engineered protein binds and activates IFNAR1 or IFNAR2 at a similar level than the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by an ISRE-GFP reporter assay or an ISRE luciferase reporter assay. In some embodiments, the engineered protein reduces an extracellular level of a pro-inflammatory cytokine, as compared to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by ELISA. In some embodiments, the pro-inflammatory cytokine is a chemokine, an interferon (IFN), an interleukin (IL), a lymphokine, a tumor necrosis factor (TNF), or any combination thereof. In some embodiments, the pro-inflammatory cytokine is IL-ip, IL-6, IL-8, IL-9, IL-12, IL-15, IL-17, IL-18, IFN-y, TNF-a, TNF-P, or any combination thereof. In some embodiments, the engineered protein increases an extracellular level of an anti-inflammatory cytokine, as compared to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. as measured by ELISA. In some embodiments, the anti-inflammatory cytokine is a chemokine, an IFN, an IL, a lymphokine, a TNF, a Transforming growth factor (TGF), or any combination thereof. In some embodiments, the anti-inflammatory cytokine is IL-4, IL-10, IL-11, IL-13, IFN-a, TGF-P, or any combination thereof. In some embodiments, the engineered protein binds to IFNAR1 or IFNAR2 subunits at a level less than the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by ELISA. In some embodiments, the engineered protein is an antagonist of IFNAR1 or IFNAR2 subunits. In some embodiments, the engineered protein binds and inhibits an activity of the IFNAR1 or IFNAR2 at a level greater than wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by an ISRE GFP reporter assay or an ISRE luciferase reporter assay. In some embodiments, the engineered protein is a recombinant IFN-beta or a portion thereof. In some embodiments, the engineered protein is glycosylated, carboxylated, hydroxylated, sulfated, phosphorylated, albuminated, conjugated to a polyethylene glycol (PEG) moiety, or any combination thereof. In some embodiments, a glycosylation site is introduced in the engineered protein that results in diminished binding of an antibody to the engineered protein, relative to an amount of binding of the antibody to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the modification results in a reduction of immunogenicity levels when administered to a subject, as compared to an immunogenicity level when the wildtype IFN-beta with the amino acid sequence of SEQ ID NO: 1 is administered to a subject. In some embodiments, the reduction of immunogenicity levels comprises a reduced level of anti-drug antibody in the subject when the engineered protein is administered to the subject, as compared a level of anti-drug antibody when the wildtype IFN-beta with the amino acid sequence of SEQ ID NO: 1 is administered, as measured by ELISA on a sample obtained from the subject. In some embodiments, the engineered protein, when administered to a subject in need thereof results in a reduction in a level of inflammation in a subject, relative to a level of inflammation in the subject prior to the administering. In some embodiments, the reduction in the level of inflammation is determined by analyzing levels of an autoantibody, a C reactive protein (CRP), a proteolytic en / yme. an inflammatory mediator, a marker of ongoing inflammation, or any combination thereof, as measured by ELISA. In some embodiments, the reduction in the level of inflammation in the subject results in a reduction in serum levels of C reactive protein as measured by high-sensitivity C-reactive protein (hs-CRP) test. In some embodiments, the reduction in serum levels of C reactive protein comprises a plasma concentration in the subject of from about 0.1 mg / dL to about 2.9 mg / dL. In some embodiments, the administering to the subject results in a reduction in blood levels of a pro-inflammatory cytokine as compared to the blood levels of one or more pro-inflammatory cytokines before the administering, as measured by ELISA. In some embodiments, the functional fragment thereof comprises at least: 80%, 85%, 90%, 91%, 92%, 93%, 94% 95%, 96%, 97%, 98%, 99%, or 100% sequence identity7 to any one of SEQ ID NO: 2 - SEQ ID NO: 208. In some embodiments, the functional fragment thereof comprises at least: 80%, 85%, 90%, 91%, 92%, 93%, 94% 95%, 96%, 97%, 98%, 99%, or 100% sequence length to any one of SEQ ID NO: 2 -SEQ ID NO: 208. In some embodiments, the functional fragment thereof comprises at least: 80%, 85%, 90%, 91%, 92%, 93%, 94% 95%, 96%, 97%, 98%, 99%, or 100% sequence homology7 to any one of SEQ ID NO: 2 - SEQ ID NO: 208. In some embodiments, the engineered protein comprises an amino acid sequence that comprises a natural amino acid, or a non-natural amino acid.

[0004] Also provided herein are methods of making a engineered IFN-beta protein in a cell, comprising introducing into the cell a vector encoding the engineered IFN-beta protein disclosed herein. In some embodiments, the introducing results in expression of the engineered IFN-beta protein in the cell. In some embodiments, the engineered protein is isolated from the cell. In some embodiments, the vector comprises a mammalian expression plasmid. In some embodiments, the engineered IFN-beta protein comprises a tag. In some embodiments, the tag is CBP, FLAG, GST, Myc, poly-His, or V5. In some embodiments, the tag comprises a cleavable tag or a non-cleavable tag.

[0005] Provided herein are polynucleotides encoding an engineered interferon beta (IFN-beta) protein or a functional fragment thereof, comprising a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1. In some embodiments, the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208. In some embodiments, the polynucleotide comprises at least: 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of SEQ ID NO: 209 - SEQ ID NO: 342. In some embodiments, the polynucleotide comprises any one of SEQ ID NO: 209 - SEQ ID NO: 342. In some embodiments, the polynucleotide comprises at least: 80%, 85%, 90%, 91%, 92%. 93%. 94%. 95%. 96%. 97%. 98%. 99%. or 100% sequence length to any one of SEQ ID NO: 209 - SEQ ID NO: 342. In some embodiments, each codon in the polynucleotide can independently be replaced by a degenerate codon. In some embodiments, the polynucleotide comprise one or more degenerate codons. In some embodiments, the polynucleotide comprises a nucleic acid mimic. In some embodiments, the nucleic acid mimic comprises a phosphorothioate nucleic acid, a phosphoramidate nucleic acid, a morpholino nucleic acid, a hexitol nucleic acid (HNA), a peptide nucleic acid (PNA), or a locked nucleic acid (LNA). In some embodiments, one or more codons in the polynucleotide is modified. In some embodiments, one or more residues of an amino acid in the engineered IFN-beta protein is substituted for a non-natural amino acid. Also provided herein are vectors comprising the polynucleotides disclosed herein.

[0006] Also provided herein are pharmaceutical compositions in unit dose form comprising: (a) the engineered proteins or the polynucleotides disclosed herein; and a pharmaceutically acceptable excipient, earner, or diluent. In some embodiments, the pharmaceutical composition is encapsulated. In some embodiments, the pharmaceutical composition is in the form of a liquid. In some embodiments, the pharmaceutical composition is formulated for local, systemic, or topical administration. In some embodiments, the pharmaceutical composition is formulated for oral, nasal, pulmonary, buccal, transdermal, subcutaneous, intraduodenal. enteral, parental, intravenous, or intramuscular administration. In some embodiments, the pharmaceutical composition is formulated for controlled-release. In some embodiments, the pharmaceutical composition is formulated for single-dosage administration.

[0007] Also provided herein are methods of treating a disease in a subject, the methods comprising: administering to the subject a therapeutically effective amount of a composition comprising (a) an engineered interferon beta (IFN-beta) protein or a functional fragment thereof; and (b) a pharmaceutically acceptable excipient, carrier, or diluent. In some embodiments, the engineered IFN-beta protein comprises a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1. In some embodiments, the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208. In some embodiments, the disease is an infection. In some embodiments, the infection is a viral infection. In some embodiments, the viral infection is chronic Hepatitis B infection, or a chronic Hepatitis C infection. In some embodiments, the disease is a demyelinating disorder. In some embodiments, the demyelinating disorder is selected from the group consisting of Multiple Sclerosis (MS), Acute Disseminated Encephalomyelitis (ADEM), Acute Hemorrhagic Leucoencephalitis (AHLE), Balo’s disease (Concentric Sclerosis), Charcot-Marie-Tooth disease (CMT). Guillain-Barre Syndrome (GBS), HTLV-I Associated Myelopathy (HAM), and Neuromyelitis Optica (Devic’s Disease). In some embodiments, the MS is relapsing remitting MS, non-relapsing MS, clinically isolated syndrome, and secondary progressive forms. In some embodiments, the subject is a human subject. In some embodiments, the administering is by parenchymal injection, intra-thecal injection, intra-ventricular injection, intra-cisternal injection, intratumoral injection, subcutaneous injection, intraperitoneal injection, a surgical route, or any combination thereof. In some embodiments, the administering occurs from about once a day, about twice a day, about once a week, about twice a week, about once every two weeks, about once a month, about once every7 3 months, about once every 6 months, about once every7 9 months, to about once a year. In some embodiments, the administering is at a dose selected from the group consisting of about 50 mg. about 75 mg. about 100 mg. about 150 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg, about 1000 mg, about 1200 mg, about 1500 mg, or about 2000 mg. In some embodiments, the administering is at a dose selected from the group consisting of 1 mg / kg. 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg. and 10 mg / kg. In some embodiments, the administering reduces a volume of a tumor in the subject. In some embodiments, the administering inhibits or diminishes the grow th a tumor in the subject. In some embodiments, the methods further comprise administering a second therapeutic. In some embodiments, the second therapeutic is an antibody, a peptide, an antibody drug conjugate, a CAR-T, a cancer vaccine, or any combination thereof. In some embodiments, the methods further comprise administering an immunotherapy. In some embodiments, the immunotherapy' comprises a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor is a PD-1, a PD-L1, CTLA-4 inhibitor. In some embodiments, the administering induces activation of an IFN-stimulated response element (ISRE) or a Gamma interferon activation site (GAS) element, w herein the activation of the ISRE or GAS induces an interferon-like activity7 as measured by a luciferase reporter assay.

[0008] Also provided herein are methods of treating a disease or a condition in a subject, the methods comprising: administering to the subject a therapeutically effective amount of engineered proteins disclosed herein, the polynucleotides disclosed herein, or the pharmaceutical composition disclosed herein, thereby treating the subject.

[0009] Provided herein are methods of treating a demyelinating disorder, the methods comprising: administering to the subject a therapeutically effective amount of a composition comprising: (a) an engineered interferon beta (IFN-beta) protein or a functional fragment thereof, comprising a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1, wherein the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208; and (b) a pharmaceutically acceptable excipient, carrier or diluent. In some embodiments, the demyelinating disorder is selected from the group consisting of Multiple Sclerosis (MS), Acute Disseminated Encephalomyelitis (ADEM), Acute Hemorrhagic Leucoencephalitis (AHLE), Balo’s disease (Concentric Sclerosis), Charcot-Marie-Tooth disease (CMT), Guillain-Barre Syndrome (GBS), HTLV-I Associated Myelopathy (HAM), and Neuromyelitis Optica (Devic’s Disease). In some embodiments, the demyelinating disorder is a MS, wherein the MS is relapsing remitting MS, non-relapsing MS, clinically isolated syndrome, and secondary progressive forms.

[0010] Also provided herein are kits comprising the engineered proteins disclosed herein, the polynucleotides disclosed herein, or the pharmaceutical composition disclosed herein,, and a container. INCORPORATION BY REFERENCE

[0011] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The novel features of the invention are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the invention are utilized, and the accompanying drawings of which:

[0013] FIG. 1 depicts the design and application of the high-throughput molecular function assay system for assessing mutational effects on gene activity (MEGA) and to produce MEGAmaps.

[0014] FIG. 2 illustrates the application of the high-throughput molecular function assay system to screen variants of IFN-beta for biosimilar activity.

[0015] FIG. 3 depicts an overview of the high-throughput molecular function assay system to assess tethered IFN-beta autocrine signaling.

[0016] FIG. 4 depicts a graph flow-cytometry profile of ISRE reporter activity in IFN-beta treated cells vs. untreated. Untreated HEK-293 interferon-responsive reporter cells is used as control (grey; untreated) to compare with HEK-293 cells treated with purified wildtype IFN(3 ligand (dark grey; treated). The x-axis depicts relative level fluorescence of ZsGreen and the y-axis depicts the number events (cell counts).

[0017] FIG. 5 depicts a schematic of the purification of HIS-tagged engineered protein as described herein.

[0018] FIGs. 6A-6B depicts a schematic for determination of EC50 of IFN-beta signaling in engineered protein as described herein. FIG 6A. depicts a 96-well plate assay for measuring EC50 of IFN signaling for comparing IFN chimera. FIG 6B. Dose response curve output from the 96-well ISRE reporter assay.

[0019] FIG. 7 depicts an assay to determine immunogenicity of IFN leads using Peripheral Blood Mononuclear Cells (PBMCs).

[0020] FIG. 8A-8B depicts a schematic of the purification of un-tagged engineered protein as described herein. FIG. 8A depicts a schematic of the purification. FIG. 8B depicts amino acid sequence of 6xHIS and the Enterokinase Light Chain Cleavage Site (EKCS) protease site upstream of the engineered protein described herein, with the cleavage position of enterokinase indicated by a dashed vertical line.

[0021] FIG. 9 depicts the dose response of the engineered proteins described herein to determine the EC50 values in an antiviral Cytopathic Effect (CPE) assay.

[0022] FIG. 10 illustrates a schematic to determine biomarker expression in human peripheral blood mononuclear cells (PBMCs) after treatment with the engineered proteins described herein.

[0023] FIG. 11 depicts the prophylactic assessment of engineered proteins in Experimental Autoimmune Encephalomyelitis (EAE) mouse model for Multiple Sclerosis (MS).

[0024] FIG. 12 depicts the liver toxicity assessment of engineered proteins described herein in mice.

[0025] FIG. 13 depicts the pharmacokinetic study of the engineered proteins described herein in humans.

[0026] FIG. 14 depicts the immunogenicity study of the engineered proteins described herein in humans.

[0027] FIGs. 15A-15N depict the verification of the engineered proteins described herein on SDS-PAGE.

[0028] FIGs. 16A-16L depict the plate reader output after cells were treated with exemplary engineered HIS-tagged engineered IFN-beta proteins or HIS-tagged wildtype IFN-beta protein. The x-axis depicts the ligand concentration, and the y-axis depicts the ISRE reporter activity.

[0029] FIGs. 17A-17G depict the flow cytometry measurements in cells treated with exemplary HIS-tagged engineered IFN-beta proteins increased activity (GOF variant) as compared to wildtype at different concentrations. The x-axis depicts the ISRE reporter activity, and the y-axis depicts the cell counts.

[0030] FIGs. 18A-18F depict quantitative RT-PCR determination of OAS1 mRNA expression in cells in expressing exemplary HIS-tagged engineered IFN-beta proteins as compared to HIS-tagged wildtype IFN beta protein. The x-axis ligand concentration (ng / mL), and the y-axis depicts OAS1 mRNA (normalized to HPRT). DETAILED DESCRIPTION OF THE INVENTION

[0031] Unless defined otherwise, all terms of art, notations and other technical and scientific terms or terminology used herein are intended to have the same meaning as is commonly understood by one of ordinary skill in the art to which the claimed subject matter pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a substantial difference over what is generally understood in the art.

[0032] Throughout this application, various embodiments may be presented in a range format. The description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

[0033] The singular forms '‘a”, “an”, and “the” are used herein to include plural references unless the context clearly dictates otherwise. Accordingly, unless the contrary is indicated, the numerical parameters set forth in this application are approximations that can vary7 depending upon the desired properties sought to be obtained.

[0034] The terms “determining”, “measuring”, “evaluating”, “assessing,” “assaying,” and “analyzing” are often used interchangeably herein to refer to forms of measurement and include determining if an element may be present or not (for example, detection), or the amount of an element. These terms can include quantitative and qualitative determinations. Assessing can be alternatively relative or absolute. “Detecting the presence of’ includes determining the amount of something present, as well as determining whether it may be present or absent.

[0035] The term “substantially” refers to a qualitative condition that exhibits at least 70 % of a total range or degree of a feature or characteristic of interest.

[0036] The term “operably coupled” refers to functional linkage between a regulatory sequence and a nucleic acid sequence resulting in expression of the latter.

[0037] Unless otherwise indicated, open terms for example “contain,” “containing,” “include,” “including,” and the like mean comprising.

[0038] As used herein, the term, “about” or “approximately,” means within an acceptable error range for the particular value and includes a range of up to 10% of a given value. Where particular values are described in the application and claims, unless otherwise stated the term “about” meaning within an acceptable error range for the particular value should be assumed.

[0039] When used herein, a percentage of a material (e.g.. a biological material, an excipient, a compound) of a composition is with respect to a total weight of a composition. In some cases, a percentage of a material of a composition is with respect to a total volume of a composition. In some cases, “Percentage by weight” or “w / w” means ratio of the mass of the specified ingredient verses the mass of the entire composition (e.g., dosage unit).

[0040] The term “homology” can refer to the relationship among sequences whereby there is some extent of likeness, typically due to descent from a common ancestral sequence. Homologous sequences can share homology' based on genic, structural, functional and / or behavioral properties. Similarity of a polypeptide to a reference polypeptide is generally determined based on conservation of certain properties of amino acids (such as conservative substitutions), and the degree of similarity can be determined using known computer programs such the Bestfit program (Wisconsin Sequence Analysis Package, Version 8 for Unix, Genetics Computer Group, University Research Park, 575 Science Drive, Madison, Wis. 53711).

[0041] The term “native” or “wildtype,” in reference to interferon beta (IFN-beta), refers to biologically active, naturally-occurring IFN-beta, including biologically active, naturally -occurring IFN-beta variants. As used herein, the terms “interferon beta,” “IFN-beta,” “IFN-P” and “IFNP” are used interchangeably, and refer to a polypeptide or protein with at least 95% sequence identity to SEQ ID NO: 1.

[0042] The term includes the 166 amino acid human IFN-beta mature sequence (SEQ ID NO: 1).

[0043] The term "glycosylation" refers to post-translational protein modification in which carbohydrates, particularly glycans, are attached to functional groups of amino acids. Glycosylation is the process by which carbohydrates are converted to nitrogen on the side chain of asparagine or arginine (N-glycosylation); hydroxyl oxygens of serine, threonine, tyrosine, hydroxylysine or hydroxyproline side chains (O-glycosylation); phosphate of phosphoserine (phosphoglycosylation); or attached to a carbon of the tryptophan side chain (C-glycosylation).

[0044] The terms “non-naturally occurring protein,’' “engineered protein,” and “engineered IFN-beta protein” as used herein are used interchangeably. As used herein, the terms "engineered IFN-beta protein,” “non-naturally occurring IFN-beta polypeptides” or “non-naturally occurring IFN-beta proteins” is used interchangeably and refers to an engineered interferon-beta polypeptide or protein as disclosed herein.

[0045] The term "glycosylation motif refers to an amino-acid residue sequence that tends to be post-translationally modified by glycosylation (i.e., enzyme-catalyzed covalent linkage of one or more carbohydrate moieties) during protein biosynthesis. In some instances, “glycosylation motif as used herein refers to a consensus sequence motif that contains a site for glycosylation. In some embodiments, the site for glycosylation comprises an amino acid residue that is capable of forming a covalent linkage, wherein the covalent linkage is to a carbohydrate moiety’. In some embodiments, the covalent linkage occurs via a nitrogen atom. In some embodiments, the nitrogen atom is in a sidechain amide group of an asparagine [Asn] residue).

[0046] The term “aglycosylate” or “aglycosylated” or “aglycosylation” as used herein, refers to a protein that bypass glycosylation, thereby preventing glycosyl groups from being added to the protein.

[0047] The term “subject,” “host,” “individual,” and “patient” are as used interchangeably herein to refer to an animal, typically mammalian animals. Any suitable mammal can be administered a composition as described herein or be treated by a method as described herein. Non-limiting examples of mammals include humans, non-human primates (e.g., apes, gibbons, chimpanzees, orangutans, monkeys, macaques, and the like), domestic animals (e.g., dogs and cats), farm animals (e.g., horses, cows, goats, sheep, pigs) and experimental animals (e.g., mouse, rat, rabbit, guinea pig). Mammals can be any age or at any stage of development, for example a mammal can be neonatal, infant, adolescent, adult or in utero. In some embodiments, a subject is a human. Humans can be more than about: 1, 2, 5, 10, 20, 30, 40, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115 or about 120 years of age. Humans can be less than about: 1, 2, 5, 10, 20, 30, 40, 50, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115 or about 120 years of age. In some cases, a human can be less than about 18 years of age. In some cases, human is from about 1 week to about 5 weeks old, 1 month to about 12 months old. from about 1 year to about 20 years, from about 15 years to about 50 years, from about 40 years to about 80 years, or from about 60 years to about 110 years. In some cases, a human is more than about 18 years of age. A human may be a pediatric subject. A human may be an adult subject. A human is a child subject. A mammal such as a human is bom a male or a female. In some embodiments, a subject has or is suspected of having a disease or condition, such as a cancer. The subject is a patient, such as a patient being treated for a condition or a disease, such as a cancer. In some cases, a subject is a responder to cancer therapy. In some cases, a subject is a non-responder to a cancer therapy. A subject is predisposed to a risk of developing a condition or a disease. A subject is in remission from a condition or a disease. In some instances, a subject is healthy. A subject may be a subject in need thereof. A subject may have received a positive diagnosis of the cancer. A subject may have received a cancer therapy that failed to treat a cancer.

[0048] A “therapeutically effective amount” refers to an amount of a composition as disclosed herein with or without additional agents that is effective to achieve its intended purpose, for example to treat a disease. Individual patient needs may vary. Generally, the dosage required to provide an effective amount of the composition will vary, depending on the age, health, physical condition, sex, weight, extent of the disease of the recipient, frequency of treatment and the nature and scope of the disease or condition.

[0049] As used herein, the terms “treatment” or “treating” refers to a pharmaceutical or other intervention regimen for obtaining beneficial or desired results in the recipient. Beneficial or desired results include but are not limited to a therapeutic benefit and / or a prophylactic benefit. A therapeutic benefit refers to eradication or amelioration of one or more symptoms of an underlying disorder being treated. For example, a therapeutic benefit can comprise shrinking a tumor, at least partially inhibiting a tumor to spread, reducing the size of a tumor, inhibiting a tumor to grow, slowing tumor grow th, or any combination thereof. Also, a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement may be observed in the subject, notwithstanding that the subject may still be afflicted with the underlying disorder. A prophylactic effect includes delaying, preventing, or eliminating the appearance of a disease or condition, delaying, or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof. For a prophylactic benefit, a subject at risk of developing a particular disease, or a subject reporting one or more of the physiological symptoms of a disease may undergo a treatment disclosed herein, even though a diagnosis of this disease may not have been made.

[0050] As used herein, the term percent “identity,” in the context of two or more nucleic acid or polypeptide sequences, refers to two or more sequences or subsequences that have a specified percentage of nucleotides or amino acid residues that are the same, when compared and aligned for maximum correspondence, as measured using one of the sequence comparison algorithms described below (e.g., BLASTP and BLASTN or other algorithms available to persons of skill) or by visual inspection.

[0051] Depending on the application, the percent ‘'identity” can exist over a region of the sequence being compared, e.g., over a functional domain, or, alternatively, exist over the full length of the two sequences to be compared.

[0052] “Encoding” refers to the inherent property of specific sequences of nucleotides in a polynucleotide, such as a gene, a cDNA, or an mRNA, to serve as templates for synthesis of other polymers and macromolecules in biological processes having either a defined sequence of nucleotides (i.e., rRNA, tRNA and mRNA) or a defined sequence of amino acids and the biological properties resulting therefrom. Thus, a gene encodes a protein if transcription and translation of mRNA corresponding to that gene produces the protein in a cell or other biological system. Both the coding strand, the nucleotide sequence of which is identical to the mRNA sequence and is usually provided in sequence listings, and the non-coding strand, used as the template for transcription of a gene or cDNA, is referred to as encoding the protein or other product of that gene or cDNA.

[0053] As used herein “endogenous” refers to any material from or produced inside an organism, cell, tissue, or system.

[0054] As used herein, the term “exogenous” refers to any material introduced from or produced outside an organism, cell, tissue, or system.

[0055] The term “expression” as used herein is defined as the transcription and / or translation of a particular nucleotide sequence driven by its promoter.

[0056] “Expression vector"’ refers to a vector comprising a recombinant polynucleotide comprising expression control sequences operatively linked to a nucleotide sequence to be expressed. An expression vector comprises sufficient cis-acting elements for expression; other elements for expression is supplied by the host cell or in an in vitro expression system. Expression vectors include all those known in the art, such as cosmids, plasmids (e.g., naked or contained in liposomes) and viruses (e.g., Sendai viruses, lentiviruses, retroviruses, adenoviruses, and adeno-associated viruses) that incorporate the recombinant polynucleotide.

[0057] A '‘vector” is a composition of matter which comprises an isolated nucleic acid and which is used to deliver the isolated nucleic acid to the interior of a cell. Numerous vectors are known in the art including, but not limited to, linear polynucleotides, polynucleotides associated with ionic or amphiphilic compounds, plasmids, and viruses. Thus, the term “vector” includes an autonomously replicating plasmid or a virus. The term should also be construed to include nonplasmid and non-viral compounds which facilitate transfer of nucleic acid into cells, such as, for example, polylysine compounds, liposomes, and the like. Examples of viral vectors include, but are not limited to, Sendai viral vectors, adenoviral vectors, adeno-associated virus vectors, retroviral vectors, lentiviral vectors, and the like.

[0058] “Sequence identity” as used herein, refers to the similarity between two polymeric molecules, e.g., between two nucleic acid molecules, such as, two DNA molecules or two RNA molecules, or between two polypeptide molecules. When a subunit position in both of the two molecules is occupied by the same monomeric subunit; e.g., if a position in each of two DNA molecules is occupied by adenine, then they are the same at that position. The sequence identity between two sequences is a direct function of the number of matching positions; e.g., if half (e.g., five positions in a polymer ten subunits in length) of the positions in two sequences are similar, the two sequences have 50% sequence identity, if 90% of the positions (e.g., 9 of 10), are matched, the two sequences have 90% sequence identity. In some instances, the phrase “sequence identity” means the percentage of identical subunits at corresponding positions in two sequences (e.g., nucleic acid sequences, amino acid sequences) when the two sequences are aligned to maximize subunit matching, therefore, taking into account gaps and insertions resulting from indels.

[0059] In the context of the present invention, the following abbreviations for the commonly occurring nucleic acid bases are used. “A” refers to adenosine, “C” refers to cytosine, “G” refers to guanosine, “T” refers to thymidine, and “U” refers to uridine.

[0060] Unless otherwise specified, a “nucleotide sequence encoding an amino acid sequence” includes all nucleotide sequences that are degenerate versions of each other and that encode the same amino acid sequence. The phrase nucleotide sequence that encodes a protein or an RNA may also include introns to the extent that the nucleotide sequence encoding the protein may in some versions contain an intron(s).

[0061] The term “linked” refers to functional linkage between a regulatory sequence and a heterologous nucleic acid sequence resulting in expression of the latter. For example, a first nucleic acid sequence is operably linked with a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Generally, operably linked DNA sequences are contiguous and, where necessary to join two protein coding regions, in the same reading frame.

[0062] The term “polynucleotide” as used herein is defined as a chain of nucleotides. Furthermore, nucleic acids are polymers of nucleotides. Thus, nucleic acids and polynucleotides as used herein are interchangeable. One skilled in the art has the general knowledge that nucleic acids are polynucleotides, which is hydrolyzed into the monomeric “nucleotides.” The monomeric nucleotides is hydrolyzed into nucleosides. As used herein polynucleotides include, but are not limited to, all nucleic acid sequences which are obtained by any means available in the art, including, without limitation, recombinant means, i.e., the cloning of nucleic acid sequences from a recombinant library or a cell genome, using ordinary cloning technology and PCR™. and the like, and by synthetic means.

[0063] As used herein, the terms “peptide.” “polypeptide,” and “protein” are used interchangeably, and refer to a compound comprised of amino acid residues covalently linked by peptide bonds. A protein or peptide must contain at least two amino acids, and no limitation is placed on the maximum number of amino acids that comprise a protein’s or peptide’s sequence. Polypeptides include any peptide or protein comprising two or more amino acids joined to each other by peptide bonds. As used herein, the term refers to both short chains, which also commonly are referred to in the art as peptides, oligopeptides and oligomers, for example, and to longer chains, which generally are referred to in the art as proteins, of which there are many types. “Polypeptides” include, for example, biologically active fragments, substantially homologous polypeptides, oligopeptides, homodimers, heterodimers, variants of polypeptides, modified polypeptides, derivatives, analogs, fusion proteins, among others. The polypeptides include natural peptides, recombinant peptides, synthetic peptides, or a combination thereof.

[0064] Ranges: throughout this disclosure, various aspects of the invention are presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3. from 1 to 4, from 1 to 5, from 2 to 4. from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0065] Although this disclosure has been described in terms of certain implementations and uses, other implementations and other uses, including implementations and uses which do not provide all of the features and advantages set forth herein, are also within the scope of this disclosure. Components, elements, features, acts, or steps is arranged or performed differently than described and components, elements, features, acts, or steps is combined, merged, added, or left out in various implementations. All possible combinations and sub combinations of elements and components described herein are intended to be included in this disclosure. No single feature or group of features is necessary or indispensable.

[0066] Certain features that are described in this disclosure in the context of separate embodiments are implemented in combination in a single embodiment. Conversely, various features that are described in the context of a single implementation is also implemented in multiple embodiments separately or in any suitable sub combination. Moreover, although features may be described above as acting in certain combinations, one or more features from a claimed combination can in some cases be excised from the combination, and the combination may be claimed as a sub combination or variation of a sub combination.

[0067] While operations may be described in the specification in a particular order, such operations need not be performed in the particular order described or in sequential order, or that all operations be performed, to achieve desirable results. Other operations that are not depicted or described is incorporated in the example methods and processes. For example, one or more additional operations can be performed before, after, simultaneously, or between any of the described operations. Additionally, the operations may be rearranged or reordered in some implementations. Also, the separation of various components in the implementations described above should not be understood as requiring such separation in all implementations, and it should be understood that the described components and systems can generally be integrated together in a single product or packaged into multiple products. Additionally, some implementations are within the scope of this disclosure.

[0068] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. Overview

[0069] Disclosed herein are engineered interferon (IFN)-beta proteins, where the engineered proteins described herein are single glycosylated, double glycosylated, multi-glycosylated or aglycosylated. The engineered proteins as described herein have binding affinity to an interferon-a / (3 receptor-1 (IFNAR1) or interferon-a / p receptor-2 (IFNAR2) heterodimer subunits, which is substantially similar or greater than the binding activity of the wildtype IFN-beta protein. IFN-beta is a biologic used to reduce inflammation and treat inflammatory conditions, such as Multiple Sclerosis. These novel engineered proteins are designed to have increased serum halflife, increased solubility, increased signaling, and reduced immunogenicity.

[0070] Also disclosed herein are compositions comprising non-naturally occurring protein, methods of making the engineered protein, and methods for treating disease. In some embodiments, the engineered protein as described herein is a recombinant IFN-beta protein. In some embodiments, the engineered protein can act as a signaling agent. In some embodiments, the biological activity of the engineered IFN-beta protein is identical to wildtype IFN-beta. Glycosylated engineered proteins

[0071] Disclosed herein are engineered interferon beta (IFN-beta) proteins or functional fragments thereof, comprising a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1, wherein the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208. Also disclosed herein are engineered, non-naturally occurring interferon (IFN)-beta protein variants comprising an amino acid substitution that results in a conversion of a motif in the amino acid sequence of SEQ ID NO: 1 that is not a glycosylation motif into a glycosylation motif, thereby introducing a glycosylation motif into the engineered protein. In some embodiments, the amino acid substitution results in the conversion of a glycosylation motif in the amino acid sequence of SEQ ID NO: 1 into a motif that is not a glycosylation motif, thereby eliminating a glycosylation motif from the engineered protein. In some embodiments, the amino acid substitution introduces, eliminates or both introduces and eliminates one or more glycosylation sites in the amino acid sequence of SEQ ID NO: 1. The engineered proteins has substantially wildtype IFN-beta like biological activity. In some embodiments, the engineered proteins described herein comprise a polypeptide sequence having a sequence with at least: 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of SEQ ID NO: 2 - SEQ ID NO: 208. Such variants can be used as a treatment for inflammatory diseases or conditions such as multiple sclerosis and cancer.

[0072] In some embodiments, the engineered protein has an amino acid substitution (e.g., one or more amino acid substitutions) recited in TABLE 1 below, relative to a wildtype IFN-beta protein having an amino acid sequence of SEQ ID NO: 1 (MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQQFQKEDAALT IYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINHLKTVLEEKLEKEDFTRGKLMS SLHLKRYYGRILHYLKAKEYSHCAWTIVRVEILRNFYFINRLTGYLRN). An engineered, non-naturally occurring protein variant having an amino acid substitution recited in TABLE 1 which results in similar biological activity (e.g., binding activity’) to a wildtype IFN-beta protein. TABLE 1. Exemplary IFN-beta amino acid substitutions Amino Acid Substitution - Mutated Codon A68N-AAC & F70T-ACC jVNN-AAL^________ A89N-AAC & V91T-ACC C17N-AAC & K19T-ACC____________________ C31N-AAC & K33T-ACC____________________ D34N-AAC & M36T-ACC___________________ D39N-AAC & P41T-ACC____________________ D39T-ACC D54N-AAC_&A56T-ACC__________ D73N-AAC___________________ E104N-AAC & L106T-ACC E107N-AAC & E109T-ACC E29N-AAC & C31T-ACC__________ E42N-AAC & I44T-ACC___________ E43N-AAC & K45T-ACC__________ E53N-AAC & A55T-ACC__________ E61N-AAC & L63T-ACC E61N-AAC & L63T-ACC & N80Q-CAG E81N-AAC & I83T-ACC___________ F15N-AAC_&C17T-ACC__________ F38N-AAC & I40T-ACC___________ F50N-AAC & K52T-ACC__________ F67N-AAC & I69T-ACC___________ F67N-AAC & I69T-ACC & N80Q-CAG F67T-ACC______________________ F67T-ACC & N80Q-CAG__________ F70N-AAC & Q72T-ACC__________ F70N-AAC & Q72T-ACC & N80Q-CAG F8N-AAC & Q10T-ACC G26N-AAC & L28T-ACC__________ G78N-AAC & N80T-ACC__________ G7N-AAC & L9T-ACC H93N-AAC & I95T-ACC___________ H97N-AAC & K99T-ACC__________ I59N-AAC & E61T-ACC___________ I59N-AAC & E61T-ACC & N80Q-CAG I66N-AAC & A68T-ACC___________ I66N-AAC & A68T-ACC & N80Q-CAG I69N-AAC & R71T-ACC___________ I69N-AAC & R71T-ACC & N80Q-CAG I95N-AAC & H97T-ACC___________ K105N-AAC & E107T-ACC_________ K108N-AAC & DI10T-ACC________ K19N-AAC & L21T-ACC__________ K33N-AAC & R35T-ACC__________ K45N-AAC & L47T-ACC__________ K52N-AAC & D54T-ACC__________ K99N-AAC & V101T-ACC_________ L106N-AAC & K108T-ACC_________ L20N-AAC & W22T-ACC L21N-AAC & Q23T-ACC__________ L24N-AAC & G26T-ACC__________ L28N-AAC & Y30T-ACC L32N-AAC & D34T-ACC L47N-AAC & Q49T-ACC____________ L57N-AAC & I59T-ACC & N80Q-CAG L5N-AAC & G7T-ACC L63N-AAC_&N65T-ACC____________ L63N-AAC&N65T-ACC & N80Q-CAG L6N-AAC & F8T-ACC______________ L87N-AAC & A89T-ACC____________ L88N-AAC & N90T-ACC____________ L88T-ACC L98N-AAC______________________ L98T-ACC_______________________ L9N-AAC & RI IT-ACC_____________ M36N-AAC & F38T-ACC____________ M62N-AAC & Q64T-ACC___________ M62N-AAC & Q64T-ACC & N80Q-CAG N80Q-CAG & A89N-AAC & V91T-ACC N80Q-CAG & E81N-AAC & I83T-ACC N80Q-CAG & H93N-AAC & I95T-ACC N80Q-CAG & I83N-AAC & E85T-ACC N80Q-CAG & I95N-AAC & H97T-ACC N80Q-CAG & K105N-AAC & E107T-ACC N80Q-CAG & L102N-AAC & E104T-ACC N80Q-CAG & L87N-AAC & A89T-ACC N80Q-CAG & L88N-AAC & N90T-ACC N80Q-CAG & L88T-ACC____________ N80Q-CAG & L98N-AAC___________ N80Q-CAG & L98T-ACC____________ N80Q-CAG & Q94N-AAC & N96T-ACC N80Q-CAG & T82N-AAC & V84T-ACC N80Q-CAG & V101N-AAC & E103T-ACC N80Q-CAG & V84N-AAC & N86T-ACC N80Q-CAG & V91N-AAC & H93T-ACC N80Q-CAG & Y92N-AAC & Q94T-ACC N80Q-CAG & Y92T-ACC___________ Q10N-AAC_____________________ Q16N-AAC & Q18T-ACC Q16T-ACC_______________________ Q18N-AAC & L20T-ACC____________ Q46N-AAC & Q48T-ACC____________ Q48N-AAC & F50T-ACC____________ Q49N-AAC & Q51T-ACC____________ Q51N-AAC & E53T-ACC Q64N-AAC & I66T-ACC_____________ Q64N-AAC & I66T-ACC & N80Q-CAG Q72N-AAC RI 1N-AAC R27N-AAC & E29T-ACC___________ R27T-ACC______________________ R35N-AAC & N37T-ACC R71N-AAC & D73T-ACC___________ R71N-AAC & D73T-ACC & N80Q-CAG S12N-AAC &N14T-ACC__________ S13N-AAC &F15T-ACC___________ S2N-AAC & N4T-ACC S75N-AAC S75N-AAC & N80Q-CAG__________ S76N-AAC & G78T-ACC___________ S76N-AAC & G78T-ACC & N80Q-CAG T100N-AAC & L102T-ACC__________ T58N-AAC & Y60T-ACC___________ T58N-AAC & Y60T-ACC &N80Q-CAG T77N-AAC & W79T-ACC & N80Q-CAG V84N-AAC & N86T-ACC__________ V91N-AAC & H93T-ACC___________ W22N-AAC & L24T-ACC__________ W79N-AAC & E81T-ACC__________ Y30N-AAC & L32T-ACC Y3N-AAC & L5T-ACC____________ Y60N-AAC & M62T-ACC__________ Y60N-AAC & M62T-ACC & N80Q-CAG

[0073] In some embodiments, the amino acid substitution results in the conversion of 1, 2, 3, 4, or more motif(s) in the amino acid sequence of SEQ ID NO: 1 that are not glycosylation motifs into 1, 2, 3. 4, or more motif(s). In some embodiments, the conversion results in the introduction of a glycosylation motif into the non-naturally occurring protein. In other embodiments, the amino acid substitution results in the conversion of 1, 2, 3, 4, or more glycosylation motif(s) in the amino acid sequence of SEQ ID NO: 1 into 1, 2, 3, 4, or more motif(s) that are not glycosylation motifs. In some embodiments, the conversion results in the elimination of a glycosylation motif into the non-naturally occurring protein. In some embodiments, a single glycosylation site is introduced. In other instances, a double-glycosylation site is introduced. In some instances, the double-glycosylation site contains a native glycosylation site of wildty pe IFN-beta (Asn 80) protein. In other embodiments, the native glycosylation site of wildtype IFN-beta protein is eliminated in the non-naturally occurring protein. In some embodiments, the non-naturally occurring protein is aglycosylated.

[0074] In some embodiments, the non-naturally occurring protein as disclosed herein having an amino acid substitution as recited in TABLE 1 can be present as a salt, such as a pharmaceutically acceptable salt. In some embodiments, the pharmaceutically acceptable salt can include: acetate, acrylate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, bisulfite, bitartrate, bromide, butyrate, butyn-l,4-dioate, camphorate, camphorsulfonate, caproate, caprylate, chlorobenzoate, chloride, citrate, cyclopentanepropionate, decanoate, digluconate, dihydrogenphosphate, dinitrobenzoate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hexyne-1,6-dioate, hydroxy benzoate, y-hydroxybutyrate. hydrochloride, hydrobromide, hydroiodide, 2-hydroxy ethanesulfonate, iodide, isobutyrate, lactate, maleate, malonate, methanesulfonate, mandelate, metaphosphate, methanesulfonate, methoxybenzoate, methylbenzoate, monohydrogenphosphate, 1-napthalenesulfonate, 2-napthalenesulfonate, nicotinate, nitrate, palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, pyrosulfate, pyrophosphate, propiolate, phthalate, phenylacetate, phenylbutyrate, propanesulfonate, salicylate, succinate, sulfate, sulfite, succinate, suberate, sebacate, sulfonate, tartrate, thiocyanate, tosylate, undeconate and xylenesulfonate.

[0075] In some embodiments, the non-naturally occurring protein having an amino acid substitution recited in TABLE 1 can be encoded by a vector (e.g., a DNA vector). In some cases, the vector can be a viral vector such as a lentiviral vector or an adeno-associated viral vector.

[0076] In some embodiments, the non-naturally occurring protein variant has an amino acid substitution (e.g.. one or more amino acid substitutions) recited in TABLE 2 below, relative to a wildtype IFN-beta protein having an amino acid sequence of SEQ ID NO: 1. A non-naturally occurring protein variant having an amino acid substitution where the amino acid substitution introduces aN-linked glycosylation site recited in TABLE 2 has similar biological activity (e.g., binding activity) to a wildtype IFN-beta protein. TABLE 2. Exemplary IFN-beta amino acid substitutions Amino Acid Substitution - Mutated Codon L57G-GGC D34N-AAC & M36T-ACC D39N-AAC & P41T-ACC D54N-AAC & A56T-ACC D73N-AAC E104N-AAC & L106T-ACC E107N-AAC & E109T-ACC E42N-AAC & I44T-ACC E43N-AAC & K45T-ACC E53N-AAC & A55T-ACC E81N-AAC &I83T-ACC F15N-AAC &C17T-ACC F50N-AAC & K52T-ACC G26N-AAC & L28T-ACC G78N-AAC & N80T-ACC G7N-AAC & L9T-ACC H97N-AAC & K99T-ACC I95N-AAC & H97T-ACC kiosn^aa^&eTotgacc K108N-AAC & D110T-ACC K19N-AAC & L21T-ACC K33N-AAC & R35T-ACC K45N-AAC & L47T-ACC K52N-AAC & D54T-ACC L106N-AAC & K108T-ACC L24N-AAC & G26T-ACC L28N-AAC & Y30T-ACC L32N-AAC & D34T-ACC L47N-AAC & Q49T-ACC L57N-AAC & I59T-ACC & N80Q-CAG L5N-AAC & G7T-ACC L6N-AAC & F8T-ACC L88T-ACC L98T-ACC L9N-AAC & R11T-ACC N80Q-CAG & E81N-AAC & I83T-ACC N80Q-C AG & I83N-AAC & E85T-ACC N80Q-CAG & L88T-ACC N80Q-CAG & L98T-ACC N80Q-CAG & T82N-AAC & V84T-ACC N80Q-CAG & V84N-AAC & N86T-ACC Q10N-AAC Q16N-AAC & Q18T-ACC Q16T-ACC Q46N-AAC & Q48T-ACC Q48N-AAC & F50T-ACC Q49N-AAC & Q51T-ACC Q51N-AAC & E53T-ACC Q72N-AAC RI 1N-AAC R27N-AAC & E29T-ACC R27T-ACC R35N-AAC & N37T-ACC R71N-AAC & D73T-ACC S12N-AAC &N14T-ACC S13N-AAC & F15T-ACC S2N-AAC & N4T-ACC S75N-AAC S76N-AAC & G78T-ACC S76N-AAC & G78T-ACC & N80Q-CAG T100N-AAC & L102T-ACC V84N-AAC & N86T-ACC Y30N-AAC & L32T-ACC Y3N-AAC & L5T-ACC

[0077] In some embodiments, the engineered, non-naturally occurring protein having an amino acid substitution as recited in TABLE 2 introduces a glycosylation motif. In some embodiments, the non-naturally occurring IFN-beta is modified to comprise 1, 2, 3, 4, or more glycosylation sites than wildtype IFN-beta protein. In some embodiments, introduction of glycosylation site is accomplished by insertion of a consensus sequence motif. In some embodiments, the consensus sequence motif is a post-translational modification (PTM) consensus sequence motif as recited in TABLE 5 below. In some embodiments, the glycosylation motif has a consensus sequence motif. In some embodiments, the consensus motif is (i) N-X-S / T, where X is independently selected from any amino acid except proline; (ii) D / E-X-N-Z-S / T, where X and Z are independently-selected from any amino acid except proline; or (hi) N-X-C, where X is independently selected from any amino acid except proline.

[0078] In some embodiments, the engineered, non-naturally occurring protein having an amino acid substitution as recited in TABLE 2 introduces an amino acid substitution and can be further post-translationally modified. In some embodiments, the modifications can include: glycosylation, phosphorylation, lipidation, glycation, proline isomerization, methylation, acetylation, hydroxylation sites, and proteolysis. In some embodiments, the non-naturally occurring protein is gly cosylated, carboxylated, hydroxylated, sulfated, phosphory lated, albuminated, conjugated to a polyethylene glycol (PEG) moiety, or any combination thereof. In some embodiments, the engineered IFN-beta protein or the functional fragment thereof is glycosylated. In some embodiments, the engineered IFN-beta protein or the functional fragment thereof is double glycosylated. In some embodiments, the engineered IFN-beta protein or the functional fragment thereof is multi-glycosylated. In some embodiments, the engineered IFN-beta protein or the functional fragment thereof is aglycosylated.

[0079] In some embodiments, the engineered IFN-beta protein or a functional fragment thereof comprises at least one amino acid substitutions. In some embodiments, the at least one amino acid substitution is selected from TABLE 1 or TABLE 2. In some embodiments, the engineered IFN-beta protein comprises an amino acid sequence having at least about: 80%, 85%, 90%, 91%, 92 %, 93%, 94%, 95%, 96% 97%, 98% or 99% sequence identity to any one of SEQ ID NO: 2 -SEQIDNO: 136. TABLE 3. Exemplary engineered IFN-beta protein sequences SEQ ID NO: Mutant Amino Acid Sequence 2 K105N- AAC|E107T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQK.EDAALT1YEMLQNIFAIFRQDSSSTGWNET1VENLLANVYHQI NHLKTVLEENLTKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 3 M117N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLNSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 4 E107N- AAC|E109T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQK.EDAALT1YEMLQNIFAIFRQDSSSTGWNET1VENLLANVYHQI NHLKTVLEEKLNKTDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 5 S119N- AACH121T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSNLTLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 6 R11N-AAC MSYNLLGFLQNSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 7 L6T-ACC MSYNLTGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 8 Q10N-AAC MSYNLLGFLNRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 9 Q16T-ACC MSYNLLGFLQRSSNFTCQKLLWQLNGRLEYCLKDRMNFDIPEE1KQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 10 R27T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGTLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 11 S12N-AACN14T-ACC MSYNLLGFLQRNSTFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 12 S13N- AAC|F15T- ACC MSYNLLGFLQRSNNTQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 13 F15N-AACC17T-ACC MSYNLLGFLQRSSNNQTQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 14 Q16N-AAC|Q18T-ACC MSYNLLGFLQRSSNFNCTKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 15 K19N-AACL21T-ACC MSYNLLGFLQRSSNFQCQNLTWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 16 S2N-AAC|N4T-ACC MNYTLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 17 Q23N- AAC|N25T- ACC MSYNLLGFLQRSSNFQCQKLLWNLTGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 18 L24N-AACG26T-ACC MSYNLLGFLQRSSNFQCQKLLWQNNTRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 19 G26N-AAC|L28T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNNRTEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 20 R27N-AACE29T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGNLTYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 21 L28N-AACY30T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRNETCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 22 Y3N-AACL5T-ACC MSNNTLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 23 Y30N-AACL32T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLENCTKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 24 L32N-AAC|D34T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCNKTRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 25 K33N- AAC|R35T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLNDTMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 26 D34N-AAC|M36T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKNRTNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 27 R35N-AACN37T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDNMTFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 28 D39N-AACP41T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFNITEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 29 P41N- AACE43T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDINETIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 30 E42N- AACI44T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPNETKQ LQQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQ INHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 31 E43N-AACK45T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPENITQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 32 I44N-AACQ46T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEENKTL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 33 K45N-AAC|L47T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEINQT QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 34 Q46N-AAC|Q48T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKNL TQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 35 L47N-AACQ49T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQN QTFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 36 Q48N- AAC|F50T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL NQTQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 37 Q49N-AACQ51T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QNFTKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 38 L5N-AACG7T-ACC MSYNNLTFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 39 F50N-AAC|K52T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQNQTEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 40 Q51N-AACE53T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFNKTDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 41 K52N- AAC|D54T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQNETAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 42 E53N-AACA55T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKNDTALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 43 D54N- AACA56T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKENATLTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 44 L6N-AACF8T-ACC MSYNLNGTLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 45 G7N-AACL9T-ACC MSYNLLNFTQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 46 F8N- AAC|Q10T- ACC MSYNLLGNLTRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 47 L9N-AACR11T- ACC MSYNLLGFNQTSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 48 D73N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQNSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 49 D73N-AAC|N80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQNSSSTGWQETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 50 E103N- AAC|K105T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLNETLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 51 E104N- AAC|L106T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLENKTEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 52 E109N-AAC|F111T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKNDTTRGKLMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 53 E81N- AACI83T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNNTTVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 54 E85N- AAC|L87T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVNNTLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 55 G78N-AACN80T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTNWTETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 56 H97N-AACK99T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NNLTTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 57 I83N-AAC|E85T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETNVTNLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 58 K108N-AACD110T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLENETFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 59 K99N- AAC|V101T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLNTTLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 60 L102N- AAC|E104T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVNETKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 61 L106N- AAC|K108T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKNETEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 62 L57N- AAC|I59T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAANTTYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 63 L88T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLTANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 64 L98T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHTKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 65 N80Q-CAGE103N-AACK105T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLKTVLNETLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 66 N80Q- CAG|E104N- AACL106T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLKTVLENKTEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 67 N80Q- CAG|E107N- AACE109T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLKTVLEEKLNKTDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 68 N80Q-CAGE109N- AAC|F111T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLKTVLEEKLEKNDTTRGKLMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 69 N80Q-CAGE81N-AACI83T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQNTTVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 70 N80Q-CAGE85N- AAC|L87T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVNNTLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 71 N80Q-CAGH97N-AACK99T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NNLTTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 72 N80Q-CAGI83N- AAC|E85T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETNVTNLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 73 N80Q- CAG|K105N- AACE107T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLKTVLEENLTKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 74 N80Q- CAG|K108N- AACD110T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLKTVLEEKLENETFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 75 N80Q-CAGK99N-AAC V101T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLNTTLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 76 N80Q-CAGL102N- AAC|E104T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLKTVNETKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 77 N80Q- CAG|L106N- AACK108T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLKTVLEEKNETEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 78 N80Q-CAGL88T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLTANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 79 N80Q-CAGL98T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHTKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 80 N80Q-CAG|Q94N- AAC|N96T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHNI THLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 81 N80Q-CAGT100N-AACL102T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLKNVTEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHC A WTIVRVEILRNFYFINRLTGYLRN* 82 N80Q-CAGT82N-AACV84T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQENITENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 83 N80Q- CAG|V101N- AACE103T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQI NHLKTNLTEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 84 N80Q-CAGV84N- AAC|N86T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETINETLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 85 Q72N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRNDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 86 Q72N-AAC|N80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRNDSSSTGWQETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 87 Q94N-AAC|N96T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHNI THLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 88 R71N- AAC|D73T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFNQTSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 89 R71N- AACD73T- ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFNQTSSSTGWQETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 90 S74N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDNSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 91 S74N-AAC|N80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDNSSTGWQETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 92 S75N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSNSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 93 S75N-AAC|N80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSNSTGWQETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 94 S76N-AACG78T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSNTTWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 95 S76N- AACG78T- ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSNTTWQETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 96 T100N- AACL102T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKNVTEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 97 T58N-AACY60T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALNITEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 98 T77N-AACW79T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSNGTNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 99 T77N- AACW79T- ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSNGTQETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 100 T82N- AAC V84T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNENITENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 101 V101N- AACE103T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTNLTEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 102 V84N-AACN86T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETINETLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 103 W79N-AACE81T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGNNTTIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 104 A135N-AACE137T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKNKTYSHCA WTIVRVEILRNFYFINRLTGYLRN* 105 E137N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKNYSHC AWTIVRVEILRNFYFINRLTGYLRN* 106 F156N-AACN158T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYNITRLTGYLRN* 107 G114N- AACL116T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRNKTMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 108 G162N- AACL164T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTNYTRN* 109 H121N- AAC|K123T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLNLTRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 110 H131N- AAC|L133T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILNYTKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 111 H140N- AAC A142T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSNCT WTIVRVEILRNFYFINRLTGYLRN* 112 I145N- AACR147T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTNVTVEILRNFYFINRLTGYLRN* 113 K115N- AACM117T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGNLTSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 114 K134N-AAC|K136T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLNATEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 115 K136N- AAC Y138T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKANETSHCA WTIVRVEILRNFYFINRLTGYLRN* 116 L116N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKNMSSLHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 117 L122N-AACR124T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHNKTYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 118 L130N-AAC|Y132T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRINHTLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 119 L151N- AACN153T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEINRTFYFINRLTGYLRN* 120 L160N-AACG162T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRNTTYLRN* 121 L160T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRTTGYLRN* 122 L164N-AACN166T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYNRT* 123 R113N- AACK115T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTNGTLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 124 R124N- AAC Y126T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKNYTGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 125 R128N-AACL130T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGNITHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 126 R147N-AACE149T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVNVTILRNFYFINRLTGYLRN* 127 R159N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINNLTGYLRN* 128 S118N- AACL120T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMNSTHLKRYYGRILHYLKAKEYSHC AWTIVRVEILRNFYFINRLTGYLRN* 129 T112N- AAC|G114T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFNRTKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* 130 T144N- AAC V146T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WNITRVEILRNFYFINRLTGYLRN* 131 T161N- AAC|Y163T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLNGTLRN* 132 V148N-AACI150T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRNETLRNFYFINRLTGYLRN* 133 Y138N-AACH140T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKENSTCA WTIVRVEILRNFYFINRLTGYLRN* 134 Y155T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFTFINRLTGYLRN* 135 Y163N-AACR165T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGNLTN* 136 A89M- MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLMNVYHQI NHLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCA WTIVRVEILRNFYFINRLTGYLRN* TABLE 4. Exemplary engineered IFN-beta protein sequences SEQID NO: Mutant Amino Acid Sequence 137 Y92N-AACQ94T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVNHTINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 138 D39T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFTIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 139 C17N-AACK19T-ACC MSYNLLGFLQRSSNFQNQTLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 140 Q18N-AACL20T-ACC MSYNLLGFLQRSSNFQCNKTLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 141 L20N-AACW22T- ACC MSYNLLGFLQRSSNFQCQKNLTQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 142 L21N-AACQ23T-ACC MSYNLLGFLQRSSNFQCQKLNWTLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 143 W22N-AAC|L24T-ACC MSYNLLGFLQRSSNFQCQKLLNQTNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 144 E29N-AACC31T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLNYTLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 145 C31N- AAC|K33T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYNLTDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 146 M36N-AAC|F38T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRNNTDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 147 F38N-AACI40T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNNDTPEEIKQL QQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQIN HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TIVRVEILRNFYFINRLTGYLRN* 148 A68N-AACF70T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFNITRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 149 A68N-AACF70T-ACC|N80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFNITRQDSSSTGWQETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 150 A89N-AACV91T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLNNTYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 151 E61N-AAC|L63T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYNMTQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 152 E61N-AACL63T-ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYNMTQNIFAIFRQDSSSTGWQETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 153 F67N-AACI69T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNINATFRQDSSSTGWNETIVENLLANVYHQIN HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TIVRVEILRNFYFINRLTGYLRN* 154 F67N-AACI69T-ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNINATFRQDSSSTGWQETIVENLLANVYHQIN HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TIVRVEILRNFYFINRLTGYLRN* 155 F67T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNITAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 156 F67T-ACC|N80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNITAIFRQDSSSTGWQETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 157 F70N-AACQ72T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAINRTDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 158 F70N- AAC|Q72T- ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1NRTDSSSTGWQETIVENLLANVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 159 H93N- AAC|I95T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYNQTN HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TIVRVEILRNFYFINRLTGYLRN* 160 I59N-AACE61T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTNYTMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQIN HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TI VRVEILRNFYFINRLTGYLRN* 161 I59N- AACE61T- ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTNYTMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQIN HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TI VRVEILRNFYFINRLTGYLRN* 162 I66N-AACA68T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNNFT1FRQDSSSTGWNETIVENLLANVYHQ1N HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TIVRVEILRNFYFINRLTGYLRN* 163 I66N- AACA68T- ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNNFTIFRQDSSSTGWQETIVENLLANVYHQIN HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TIVRVEILRNFYFINRLTGYLRN* 164 I69N-AACR71T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKED AALTIYEMLQN1FANFTQD SSSTGWN ET1 VEN LLAN V YHQ1N HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TIVRVEILRNFYFINRLTGYLRN* 165 I69N- AACR71T- ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFANFTQDSSSTGWQETIVENLLANVYHQIN HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TIVRVEILRNFYFINRLTGYLRN* 166 L63N-AACN65T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMNQT1FA1FRQDSSSTGWNETIVENLLANVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 167 L63N- AAC|N65T- ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMNQTIFAIFRQDSSSTGWQETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 168 L87N-AACA89T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWNETIVENNLTNVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 169 L88N- AAC|N90T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLNATVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 170 L98N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWNETIVENLLANVYHQ1NH NKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 171 M62N-AACQ64T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYENLTNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 172 M62N-AAC|Q64T-ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYENLTNIFAIFRQDSSSTGWQET1VENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 173 N80Q- CAGA89N-AACV91T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLNNTYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 174 N80Q-CAGH93N-AAC|I95T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYNQTN HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TI VRVEILRNFYFINRLTGYLRN* 175 N80Q-CAG|I95N- AACH97T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVYHQNN TLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TI VRVEILRNFYFINRLTGYLRN* 176 N80Q-CAGL87N-AAC A89T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWQETIVENNLTNVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 177 N80Q- CAGL88N- AACN90T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLNATVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 178 N80Q-CAGL98N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWQETIVENLLANVYHQ1NH NKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 179 N80Q- CAGV91N-AACH93T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANNYTQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 180 N80Q-CAGY92N- AAC|Q94T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWQETIVENLLANVNHTINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 181 N80Q- CAGY92T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWQETIVENLLANVTHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 182 Q64N-AACI66T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLNNTFA1FRQDSSSTGWNETIVENLLANVYHQ1N HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TIVRVEILRNFYFINRLTGYLRN* 183 Q64N-AACI66T-ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLNNTFAIFRQDSSSTGWQETIVENLLANVYHQIN HLKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAW TIVRVEILRNFYFINRLTGYLRN* 184 T58N- AACY60T- ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALNITEMLQN1FA1FRQDSSSTGWQETIVENLLANVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 185 V91N-AAC|H93T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANNYTQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 186 Y60N-AACM62T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTINETLQNIFAIFRQDSSSTGWNET1VENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 187 Y60N- AAC|M62T- ACCN80Q-CAG MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTINETLQNIFAIFRQDSSSTGWQETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 188 Y92T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVTHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 189 A142N-AAC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCNWTI VRVEILRNFYFINRLTGYLRN* 190 C141N- AACW143T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWNETIVENLLANVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHNATTI VRVEILRNFYFINRLTGYLRN* 191 E149N-AACL151T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVNITRNFYFINRLTGYLRN* 192 F154N-AACF156T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWNETIVENLLANVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNNYTINRLTGYLRN* 193 I129N- AACH131T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRNLTYLKAKEYSHCAWT I VRVEILRNFYFINRLTGYLRN* 194 I150N- AACR152T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWNETIVENLLANVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVENLTNFYFINRLTGYLRN* 195 I157N- AACR159T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILRNFYFNNTLTGYLRN* 196 K123N- AAC|Y125T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWNETIVENLLANVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLNRTYGRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 197 L133N- AAC|A135T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYNKTKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 198 R152N-AACF154T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWNETIVENLLANVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI VRVEILNNTYFINRLTGYLRN* 199 S139N-AACC141T- ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYNHTAWTI VRVEILRNFYFINRLTGYLRN* 200 V146N- AAC V148T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWNETIVENLLANVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCAWTI NRTEILRNFYFINRLTGYLRN* 201 W143N-AACI145T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHYLKAKEYSHCANTT VRVEILRNFYFINRLTGYLRN* 202 Y125N-AACG127T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRNYTRILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 203 Y126N-AACR128T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVENLLANVYHQINH LKTVLEEKLEKEDFTRGKLMSSLHLKRYNGTILHYLKAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN* 204 Y132N-AACK.134T-ACC MSYNLLGFLQRSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPEEIKQLQ QFQKEDAALTIYEMLQN1FA1FRQDSSSTGWNETIVENLLANVYHQ1NH LKTVLEEKLEKEDFTRGKLMSSLHLKRYYGRILHNLTAKEYSHCAWTI VRVEILRNFYFINRLTGYLRN*

[0080] In some embodiments, the engineered IFN-beta proteins described having an amino acid sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 136 can have an increase in biological activity as compared to wildtype IFN-beta having an amino acid sequence of SEQ ID NO: 1. In some embodiments, the engineered IFN-beta proteins described having an amino acid sequence of any one of SEQ ID NO: 137 - SEQ ID NO: 204 can have a decrease in biological activity as compared to wildtype IFN-beta having an amino acid sequence of SEQ ID NO: 1. In some embodiments, the engineered IFN-beta proteins can have more than biological activity of more than 100% of the biological activity of the wildtype IFN-beta having an amino acid sequence of SEQ ID NO: 1. In some embodiments, the engineered IFN-beta proteins can have more than 90% of the biological activity of the wildtype IFN. In some embodiments, the engineered IFN-beta proteins can have more than 80% of the biological activity of the wildtype IFN. In some embodiments, the engineered IFN-beta proteins can have more than 70% of the biological activity of the wildtype IFN. In some embodiments, the engineered IFN-beta proteins can have more than 60% of the biological activity of the wildtype IFN. In some embodiments, the engineered IFN-beta proteins can have more than 50% of the biological activity of the wildty pe IFN. In some embodiments, the engineered IFN-beta proteins can have more than 40% of the biological activity of the wildtype IFN. In some embodiments, the engineered IFN-beta proteins can have more than 30% of the biological activity of the wildtype IFN. In some embodiments, the engineered IFN-beta proteins can have more than 20% of the biological activity of the wildtype IFN. In some embodiments, the engineered IFN-beta proteins can have more than 10% of the biological activity of the wildtype of which it is deduced (i.e., the wildtype IFN of which the coding sequence has been mutated to obtain the engineered IFN-beta).

[0081] In some embodiments, the engineered IFN-beta protein binds to an interferon-a / p receptor-l(IFNARl) or interferon-a / p receptor-2 (IFNAR2) heterodimer subunits at a level greater than or equal to the wildtype IFN-beta protein. In some embodiments, the binding affinity is measured by enzyme-linked immunosorbent assay (ELISA). In some embodiments, the engineered IFN-beta protein binds to an interferon-a / p receptor-1 (IFNARI) or interferon-a / p receptor-2 (IFNAR2) heterodimer subunits at a level greater than or equal to the wildtype IFN-beta protein as measured by ELISA and / or a GigaAssay e.g., measuring signaling activity to an ISRE-GFP reporter in a live human cell line in vitro assay. In some embodiments, the engineered IFN-beta protein is an agonist of IFNAR1 or IFNAR2 subunits.

[0082] In some embodiments, the engineered IFN-beta protein binds and activates IFNAR1 or IFNAR2 at a level greater than the wildtype IFN-beta protein with the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the engineered IFN-beta protein is an agonist of IFNAR1 or IFNAR2 subunits In some embodiments, the engineered IFN-beta protein binds and inhibits IFNAR1 or IFNAR2 at a level greater than the wildtype IFN-beta protein with the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the engineered IFN-beta protein is an antagonist of IFNAR1 or IFNAR2 subunits. In some embodiments, the activity7 of IFNAR1 or IFNAR2 is measured by a reporter assay. In some embodiments, the reporter assay is an Interferon Stimulated Response Element (ISRE) Green fluorescent protein (GFP) reporter assay or an ISRE luciferase reporter assay.

[0083] In some embodiments, the engineered IFN-beta protein reduces extracellular level of a pro-inflammatory7 cytokine as compared to wildtype IFN-beta protein. In some embodiments, the pro-inflammatory' cytokine is a chemokine, an interferon (IFN), an interleukin (IL), a lymphokine, a tumor necrosis factor (TNF), or any combination thereof. In some embodiments, the pro-inflammatory' cytokine is IL-ip, IL-6, IL-8, IL-9, IL-12, IL-15, IL-17, IL-18, IFN-y, TNF-a, TNF-P, or any combination thereof. In other embodiments, the engineered IFN-beta protein increases extracellular level of an anti-inflammatory7 cytokine as compared to wildtype IFN-beta protein. In some embodiments, the anti-inflammatory cy tokine is a chemokine, an IFN, an IL, a lymphokine. a TNF. a Transforming growth factor (TGF), or any combination thereof. In some embodiments, the anti-inflammatory' cytokine is IL-4, IL-10, IL-11, IL-13, IFN-a, TGF-P, or any combination thereof.

[0084] In some embodiments, the engineered IFN-beta protein having amino acid sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 136 has reduced immunogenicity when administered to a subject, as compared to administering the wildtype IFN-beta with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the reduced immunogenicity comprises a reduced level of anti-drug antibody in the subject when the engineered IFN-beta protein is administered to the subject, as compared a level of anti-drug antibody when the wildtype IFN-beta with the amino acid sequence of SEQ ID NO: 1 is administered, as measured by ELISA on a sample obtained from the subject. In some embodiments, the engineered IFN-beta protein, when administered to a subject in need thereof, reduces a level of inflammation in a subject, relative to a level of inflammation in the subject prior to the administering. In some embodiments, the reduced level of inflammation is determined by analyzing levels of an autoantibody, a C reactive protein (CRP), a proteolytic en / yme. an inflammatory mediator, a marker of ongoing inflammation, or any combination thereof, as measured by ELISA. In some embodiments, the reduced level of inflammation in the subject results in reduced serum levels of C reactive protein as measured by high-sensitivity C-reactive protein (hs-CRP) test. In some embodiments, the reduced serum levels of C reactive protein comprises a plasma concentration in the subject of from about 0.1 mg / dL to about 2.9 mg / dL. In some embodiments, the administering of the engineered IFN-beta protein as disclosed herein to the subject results in reduced blood levels of a pro-inflammatory cytokine as compared to the blood levels of one or more pro-inflammatory cytokines before the administering, as measured by ELISA.

[0085] In some embodiments, the engineered IFN-beta protein has an amino acid substitution (e.g., one or more amino acid substitutions) recited in TABLE 1 or TABLE 2 below, relative to a wildtype IFN-beta protein having an amino acid sequence of SEQ ID NO: 1. An engineered IFN-beta protein variant having an amino acid substitution recited in TABLE 1 or TABLE 2 has similar biological activity (e.g., binding activity) to a wildtype IFN-beta protein.

[0086] In some embodiments, the engineered IFN-beta protein comprises an amino acid sequence having at least about: 80%. 85%, 90%, 91%, 92 %, 93%, 94%, 95%, 96% 97%, 98% or 99% sequence identity to any one of SEQ ID NO: 137 - SEQ ID NO: 208.

[0087] In some embodiments, the non-naturally occurring protein having an amino acid substitution where the amino acid substitution results in the production of a post-translational modification (PTM) consensus sequence motif as recited in TABLE 5 below. In some embodiments, a “x” or “X"’ in the PTM consensus sequence motif as recited in TABLE 5 below can represent any amino acid.

[0088] In some embodiments, the non-naturally occurring protein having an amino acid substitution having an amino acid substitution as recited in TABLE 2 can be present as a salt, such as a pharmaceutically acceptable salt. In some embodiments, the pharmaceutically acceptable salt can include: acetate, acrylate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, bisulfite, bitartrate, bromide, butyrate, butyn-1,4-dioate, camphorate, camphorsulfonate, caproate, caprylate, chlorobenzoate, chloride, citrate, cyclopentanepropionate, decanoate, digluconate, dihydrogenphosphate, dinitrobenzoate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, glycolate, hemisulfate, heptanoate, hexanoate, hexyne-1,6-dioate, hydroxybenzoate, y-hydroxybutyrate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy ethanesulfonate, iodide, isobutyrate, lactate, maleate, malonate, methanesulfonate, mandelate, metaphosphate, methanesulfonate, methoxybenzoate, methylbenzoate, monohydrogenphosphate, 1-napthalenesulfonate, 2-napthalenesulfonate, nicotinate, nitrate, palmoate. pectinate, persulfate. 3-phenylpropionate, phosphate, picrate, pivalate, propionate, pyrosulfate, pyrophosphate, propiolate, phthalate, phenylacetate, phenylbutyrate, propanesulfonate, salicylate, succinate, sulfate, sulfite, succinate, suberate, sebacate, sulfonate, tartrate, thiocyanate, tosylate, undeconate and xylenesulfonate. TABLE 5. Exemplary IFN-beta Consensus Sequence Motif Amino Acid Motif [AG][KR] [AG]Rx[AG]_________________________ [DE] [DE] [DEL]XYXX[FMLVI] [DE] [DE]X[LIV]XXYXX[LIV] [DE]X[LIV]XYXX[LIV] [DE]x[ST] [FILMVWY]x[DE]____________ [DE]XY___________________________ [DE]YY___________________________ [DG][AV]DD_______________________ [EAD] [EA] [EA] [IEV]Y[ED] [ED] [IVE] [EIV] [ED] [ED] [ED] [ED] Y[VEI] F [IVF]__________ [ED] [ED] [EDG] [IVL] Y[GED]E[FILV] [DE] [ED][ED]XXXYVA [ED]FY[GA][FY][AG]_________________ [ED]GSG[DE]_______________________ [ED]xK[VILFP] [ED]Y[IFLV] [ED]YY[KR]________________________ [EDY]Y___________________________ [EP][FIL]YAX[FILV] [EP] [MLIVF] Y [GA]X[MLI VFY] A_______ [FWYL] x[HAT] D [ ACFGHILMNSTVWY] [ILV]ExD____________________________ [ILV]xxR[VF][GS]______________________ [ILV]xxYx[LV]_________________________ [IVL]Yx[PF] [IVL]Yxx[PF]_____________________________________________ [IVL]Yxxx[PF]____________________________________________ [IVL]Yxxxx[PF] [IVL]Yxxxxx[PF]__________________________________________ [KR][AST]K[DNQK]__________________________________ [KR]R_______________________________________________ [KR]xRxx[ST]____________________________________________ [KR]xx[DE]xxxY [KR]xx[ST]_______________________________________________ [KR]xxx[DE]xxY________________________________________ [KRHDE]C[DE]______________________________________ [KRHEQSTAG] G[FYLIVM] [ST] [IT] [LIVP]E[LIVMFWSTAG] [LN][RI]TY____________________________________________ [RK][RK][RK]x[ST]_______________________________________ [RK] [RK]x[ST]___________________________________________ [RK][RK]x[ST]___________________________________________ [RK] [RK]x[ST]___________________________________________ [RKTC][IVL]Y[TQHS]Y[IL]QSR_________________________ [ST] [ED] [FL] [FW]_____________________________________________ [ST]P______________________________________________________ [ST]P______________________________________________________ [ST]Px[KR]_______________________________________________ [ST]Q___________________________________________________ [ST]x[KR] [ST]xx[DE]_________________________________________________ [ST]xxE__________________________________________________ [ST]xxx[ST]________________________________________________ [ST]xxx[ST]________________________________________________ [ST]xxx[ST]________________________________________________ [ST]xxx[ST] [ST]xxx[ST]________________________________________________ [ST]xxx[ST]________________________________________________ [ST]xxx[ST]________________________________________________ [ST]xxx[ST]________________________________________________ [ST]xxxS___________________________________________________ [VILAFP]KxE_______________________________________ [VILMFPC]KxE____________________________________ [WL]EHD_______________________________________ [WL]EHD_______________________________________ [WL]EHD[ACFGHILMNSTVWY]_____________________ [YL]EVD_________________________________________ [YL]EVD [YW]EVD_____________________________________ <G[DERKHPFYW]xx[STAGCN][ACDEFGHIKLMNQRSTVWY] <MG[CGNQST]xx[STAGCN][ACDEFGHIKLMNQRSTVWY] <MGxxx[ST] A[EA]EEY[FV]F[LFMIV]F___________________________ AEV[IVLF]YAA[PF]F_______________________________ C [AVLIFYWM] [AVLIFYWM] [ACDEFGHIKNPQRSTVWY]> C [ AVLIFYWM] [ AVLIFYWM] [LM] >__________________ C [AVLIFYWM] [AVLIFYWM]x>____________________ C[CFGHIKLMPRSTVWY] [LIVM]x>__________________ C[CFGHIKLMPRSTVWY] [LIVM]x>__________________ Cx[DN]xxxx[FY]xCxC________________________________ Cx [DN] xxxx[FY] xCxC CxSxPC CxxM>______________________________________ Cxxx[ST]C___________________________________________ D[ED] [EDG] [IVL]Y[GE]E[FI]F__________________________ DDDDK[ACDEFGHIKLMNQRSTVWY]______________ DDDDxxS___________________________________ DDDxxS_____________________________________ DEVD[ACFGHILMNSTVWY]_____________________ DEVD[ACFGHILMNSTVWY]_____________________ DEVD[ACFGHILMNSTVWY]_____________________ DGADD________________________________ DGADD DGADD________________________________ DGVDD________________________________ DGVDD DGVDD________________________________ DGVDD________________________________ DxxIYE__________________________________________ DxxSIIxFF___________________________________________ DYYR_________________________________ DYYR E[DFY]YM___________________________________ E[FDY]Y_______________________________________ E[FILMTVWY]TYGx[FILMVWY]___________________ EE[DE]IYFFFF____________________________________ EE[END]Y[MF] [MF] [MFIE] [MF]_______________________ EEEIY[GE]EFD__________________________________ EEEYMMMM__________________________ EEIYYYVH_________________________________ EFY[AG]TY[AG]_________________________________ EFYAX[VI]G[RKH]S______________________________ ENLYFQ[GS]____________________________________ ERIYARTK ETVRFQ[GS]____________________________________ EX[IVLF]Y[GA]V[LVFI][FLVI]________________________ ExIYGVLF EXIYXXPX ExxYxQ[GS]_____________ ExxYxQS_______________ ExYxF F[MLVI]Y______________ FEADG______________ FxRxxSFF_______________ FxxF[ST][FY]______________ FxxF[ST][FY]> G[KR][KR] GPx___________________ GPxSPxP________________ IETD[ACFGHILMNSTVWY] IYAxP__________________ IYAxP__________________ IYGxL_________________ KAGSRGLTS__________ KKRxxS_______________ KxxxRxxTxL KxxxSPxHR LETD[ACFGHILMNSTVWY] LGxGD LxRQMS______________ LxRxx[ST] LxRxx[ST] LxRxx[ST] LxRxx[ST] Nx[TS] Px[ST]_____________________ Px[ST]P___________________ PxTP QPSxxE________________ R[KR]x[ST]_______________ R[KR]x[ST]_______________ R[KR]x[ST]_______________ RDXYXTDYYR________ RKKx[ST]_______________ RKxSF________________ RKxSFK______________ RKxSFK______________ RQxSFD______________ RRFxSLR______________ RRKxSxKR RRxSI___________________ RRxSR_______________ Rx[KR]R Rx[RK]R Rx[ST]_____________________ Rx[ST]_____________________ Rx[ST]_____________________ RxGSF_________________ RxRxx[ST] RxRxx[ST] Rxx[ST] Rxx[ST]V________________ RxxR RxxRR RxxRxS RxxSFF RxxSFRR SGxG_________________ SxD____________________ Sxx[ST]_____________________ SxxxS T[FL]CGT TxY__________________ VEHD[ACFGHILMNSTVWY] VEID[ACFGHILMNSTVWY] WxxW______________ WxxW______________ YASI__________________ YIDL__________________ YMXM____________ YMXM____________ YMXM____________ Yxx[LIV]_________________ YXX[LIV] YxxQ_________________ YxxxYY

[0089] In some embodiments, the non-naturally occurring protein having an amino acid substitution recited in TABLE 5 can be encoded by a vector (e.g., a DNA vector). In some cases, the vector can be a viral vector such as a lentiviral vector or an adeno-associated viral vector.

[0090] In some embodiments, the non-naturally occurring protein as disclosed herein is a recombinant IFN-beta protein. In some embodiments, the non-naturally occurring protein binds to an interferon-a / p receptor-l(IFNARl) or interferon-a / p receptor-2 (IFNAR2) heterodimer subunits at a level greater than or equal to the wildtype IFN-beta protein. In some embodiments, the binding affinity is measured by enzyme-linked immunosorbent assay (ELISA). In some embodiments, the non-naturally occurring protein binds to an interferon-a / p receptor-1(IFNARI) or interferon-a / p receptor-2 (IFNAR2) heterodimer subunits at a level greater than or equal to the wildtype IFN-beta protein as measured by ELISA and / or a GigaAssay e.g., measuring signaling activity to an ISRE-GFP reporter in a live human cell line in vitro assay. In some embodiments, the non-naturally occurring protein is an agonist of IFNAR1 or IFNAR2 subunits. In other embodiments, the non-naturally occurring protein binds and activates IFNAR1 or IFNAR2 at a level substantially similar to the wildtype IFN-beta protein with the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the non-naturally occurring protein binds and activates IFNAR1 or IFNAR2 at a level greater than the wildtype IFN-beta protein with the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the non-naturally occurring protein is an agonist of IFNAR1 or IFNAR2 subunits In some embodiments, the non-naturally occurring protein binds and inhibits IFNAR1 or IFNAR2 at a level greater than the wildtype IFN-beta protein with the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the non-naturally occurring protein is an antagonist of IFNAR1 or IFNAR2 subunits. In some embodiments, the activity of IFNAR1 or IFNAR2 is measured by a reporter assay. In some embodiments, the reporter assay is an Interferon Stimulated Response Element (ISRE) Green fluorescent protein (GFP) reporter assay or an ISRE luciferase reporter assay.

[0091] In some embodiments, the non-naturally occurring protein reduces extracellular level of a pro-inflammatory cytokine as compared to wildtype IFN-beta protein. In some embodiments, the pro-inflammatory cytokine is a chemokine, an interferon (IFN), an interleukin (IL), a lymphokine, a tumor necrosis factor (TNF), or any combination thereof. In some embodiments, the pro-inflammatory cytokine is IL-1 (3, IL-6, IL-8, IL-9, IL-12. IL-15, IL-17, IL-18, IFN-y, TNF-a. TNF-p. or any combination thereof. In other embodiments, the non-naturally occurring protein increases extracellular level of an anti-inflammatory cytokine as compared to wildtype IFN-beta protein. In some embodiments, the anti-inflammatory cytokine is a chemokine, an IFN, an IL, a lymphokine, a TNF, a Transforming growth factor (TGF), or any combination thereof. In some embodiments, the anti-inflammatory cytokine is IL-4, IL-10, IL-11. IL-13, IFN-a, TGF-p, or any combination thereof.

[0092] In some embodiments, a glycosylation site is introduced in the non-naturally occurring protein that results in diminished binding of an antibody to the non-naturally occurring protein relative to an amount of binding of the antibody to the wildtype IFN-beta protein. In some embodiments, the non-naturally occurring beta protein has reduced immunogenicity when administered to a subject, as compared to administering the wildtype IFN-beta with the amino acid sequence of SEQ ID NO: 1. In some embodiments, the reduced immunogenicity comprises a reduced level of anti-drug antibody in the subject when the non-naturally occurring protein is administered to the subject, as compared a level of anti-drug antibody when the wildtype IFN- beta with the amino acid sequence of SEQ ID NO: 1 is administered, as measured by ELISA on a sample obtained from the subject. In some embodiments, the non-naturally occurring beta protein, when administered to a subject in need thereof, reduces a level of inflammation in a subject, relative to a level of inflammation in the subject prior to the administering. In some embodiments, the reduced level of inflammation is determined by analyzing levels of an autoantibody, a C reactive protein (CRP), a proteolytic enzy me, an inflammatory mediator, a marker of ongoing inflammation, or any combination thereof, as measured by ELISA. In some embodiments, the reduced level of inflammation in the subject results in reduced serum levels of C reactive protein as measured by high-sensitivity C-reactive protein (hs-CRP) test. In some embodiments, the reduced serum levels of C reactive protein comprises a plasma concentration in the subject of from about 0.1 mg / dL to about 2.9 mg / dL. In some embodiments, the administering of the non-naturally occurring protein as disclosed herein to the subject results in reduced blood levels of a pro-inflammatory cytokine as compared to the blood levels of one or more pro-inflammatory' cytokines before the administering, as measured by ELISA. Polynucleotides encoding engineered IFN-beta proteins

[0093] Provided herein are polynucleotides encoding the engineered IFN-beta proteins disclosed herein. In some embodiments, the polynucleotide disclosed herein encodes an engineered interferon beta (IFN-beta) protein or a functional fragment thereof, comprising a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1, wherein the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 -SEQ ID NO: 208. Also provided herein are nucleic acids encoding the engineered IFN-beta proteins disclosed herein. In some embodiments, the nucleic acid sequence comprise a sequence at least: 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% similar to any one of SEQ ID NO: 2 - SEQ ID NO: 208. In some cases, the nucleic acid is a nucleic acid construct. In some embodiments, a nucleic acid construct comprises a polynucleotide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208. The polynucleotide of claim 55, wherein polynucleotide sequence at least: 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of SEQ ID NO: 209 - SEQ ID NO: 342. In some embodiments, the polynucleotide sequence comprises a nucleotide sequence derived from a full length engineered protein, or a fragment thereof.

[0094] In some embodiments, each codon in the polynucleotide can independently be replaced by a degenerate codon. In some embodiments, the polynucleotide sequence comprise one or more degenerate codons. In some embodiments, the polynucleotide comprises a nucleic acid mimic. In some embodiments, the nucleic acid mimic comprises a phosphorothioate nucleic acid, a phosphoramidate nucleic acid, a morpholino nucleic acid, a hexitol nucleic acid (HNA), a peptide nucleic acid (PNA), or a locked nucleic acid (LNA). In some embodiments, each codon independently codes for one or more amino acids. In some embodiments, the one or more amino acids comprise alanine, arginine, aspartic acid, glutamine, glutamic acid, glycine, praline, serine, leucine, cysteine, valine, lysine, methionine, tryptophan, phenylalanine, arginine, tyrosine, threonine, isoleucine, histidine, lysine and asparagine. In some embodiments, one or more codons in the polynucleotide is modified.

[0095] In some embodiments, one or more residues of an amino acid in the engineered protein is substituted for a non-natural amino acid. Non-limiting examples of non-natural amino acids include azidohomoalanine, homoproparglyglycine, p-bromophenylalanine, p-iodophenylalanine, azidophenylalanine, acetylphenylalanine and ethynylephenylalanine. In some embodiments, the polynucleotide encodes a polypeptide comprising a non-natural amino acid. In some embodiments, a chemical moiety is added to the non-natural amino acid. In some embodiments, the chemical moiety comprises cytotoxins, pharmaceutical drugs, dyes or fluorescent labels, a nucleophilic or electrophilic group, a ketone or aldehyde, azide or alkyne compounds, photocaged groups, tags, a peptide, a polypeptide, a protein, an oligosaccharide, poly(ethylene) glycol with any molecular weight and in any geometry, polyvinyl alcohol, metals, metal complexes, polyamines, imidizoles, carbohydrates, lipids, biopolymers, particles, solid supports, a polymer, a targeting agent, an affinity group, any agent to which a complementary reactive chemical group can be attached, biophysical or biochemical probes, isotypically-labeled probes, spin-label amino acids, fluorophores, ary l iodides and bromides. In some cases, the non-natural amino acid residue is fluorinated, electroactive or unsaturated. In some embodiments, the polypeptide further comprises a sequence encoding a linker, or a tag.

[0096] In some embodiments, the engineered IFN-beta protein as disclosed herein is encoded by a vector (e.g., a DNA vector). In some cases, a vector comprise the nucleic acids or nucleic acid constructs disclosed herein. In some cases, the vector is a viral vector such as a lentiviral vector or an adeno-associated viral vector. In some embodiments, the engineered IFN-beta protein as disclosed herein is encoded by a nucleic acid sequence. TABLE 6. Polynucleotide sequences encoding exemplary engineered proteins SEQ ID NO: Mutant Sequence 209 K105N- AACE10 7T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAACCTGACCAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 210 M117N- AAC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCAACAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 211 E107N-AACE10 9T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGAACAAGACCGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 212 S119N- AACH12 1T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAACCTGACCCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 213 RI 1N- AAC ATGAGCTACAACTTGCTTGGATTCCTACAAAACAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 214 L6T-ACC ATGAGCTACAACTTGACCGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 215 Q10N- AAC ATGAGCTACAACTTGCTTGGATTCCTAAACAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 216 Q16T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTACC TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 217 R27T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGACCCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 218 S12N- AACN14 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAACAGCACCTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 219 S13N- AACF15 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAACAATACCCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 220 F15N- AAC|C17 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATAACCAG ACCCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 221 Q16N-AAC|Q18 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTAAC TGTACCAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 222 K19N- AACL21 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAACCTCACCTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 223 S2N-AACN4T -ACC ATGAACTACACCTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 224 Q23N- AAC|N25 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGAACCTTACCGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 225 L24N- AACG26 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAAAACAATACCAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 226 G26N- AACL28 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATAACAGGACCGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 227 R27N- AACE29 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAACCTTACCTACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 228 L28N- AAC|Y30 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGAACGAAACCTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 229 Y3N-AACL5T-ACC ATGAGCAACAACACCCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 230 Y30N-AACL32 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAAAACTGCACC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 231 L32N- AACD34 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCAAC AAGACCAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 232 K33N- AACR35 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AACGACACCATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 233 D34N- AAC|M36 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGAACAGGACCAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 234 R35N- AAC|N37 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAACATGACCTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 235 D39N- AACP41 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTAACATCACCGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 236 P41N- AACE43 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCAACGAGACCATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 237 E42N-AACI44T -ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTAACGAGACCAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 238 E43N- AAC|K45 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGAACATTACCCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 239 I44N- AAC|Q46 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGAACAAGACCCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 240 K45N- AACL47 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAACCAGACCCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 241 Q46N-AAC|Q48 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGAACCTGAC CCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 242 L47N-AAC|Q49 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGAACCA GACCTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 243 Q48N- AAC|F50 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGAA CCAGACCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCC AGAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGA ATGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAA ACCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTC ACCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGG GAGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTG GACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAG ACTTACAGGTTACCTCCGAAACTGA 244 Q49N-AAC|Q51 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GAACTTCACCAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 245 L5N- AAC|G7T -ACC ATGAGCTACAACAACCTTACCTTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 246 F50N- AAC|K52 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGAACCAGACCGAGGACGCCGCATTGACCATCTATGAGATGCTCC AGAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGA ATGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAA ACCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTC ACCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGG GAGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTG GACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAG ACTTACAGGTTACCTCCGAAACTGA 247 Q51N- AACE53 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCAACAAGACCGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 248 K52N- AACD54 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAACGAGACCGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 249 E53N- AAC|A55 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGAACGACACCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 250 D54N- AACA56 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGAACGCCACCTTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 251 L6N-AACF8T-ACC ATGAGCTACAACTTGAACGGAACCCTACAAAGAAGCAGCAATTTTCA GTGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCT C AAGGAC AGGATGAACTTTGACATCCCT GAGGAG ATT AAGCAGCTGC AGCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCC AGAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGA ATGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAA ACCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTC ACCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGG GAGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTG GACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAG ACTTACAGGTTACCTCCGAAACTGA 252 G7N-AACL9T-ACC ATGAGCTACAACTTGCTTAACTTCACCCAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 253 F8N- AAC|Q10 T-ACC ATGAGCTACAACTTGCTTGGAAACCTAACCAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 254 L9N- AACR11 T-ACC ATGAGCTACAACTTGCTTGGATTCAACCAAACCAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 255 D73N- AAC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAAACTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 256 D73N- AAC|N80 Q-CAG ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAAACTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 257 E103N-AAC|K10 5T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGAACGAGACCCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 258 E104N- AACL10 6T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGAACAAGACCGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 259 E109N- AACF111 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGAACGACACCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 260 E81N-AACI83T -ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TAACACTACCGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 261 E85N- AACL87 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTAACAACACCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 262 G78N- AAC|N80 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTAACTGGAC CGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 263 H97N- AAC|K99 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CAACCTGACCACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 264 I83N- AACE85 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTAACGTTACCAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 265 K108N- AAC|D11 OT-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAACGAGACCTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 266 K99N- AAC|V10 1T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAACACAACCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 267 L102N- AACE10 4T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCAACGAGACCAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 268 L106N-AAC|K10 8T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGAACGAGACCGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 269 L57N-AACI59T -ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCAAACACCACCTATGAGATGCTCC AGAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGA ATGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAA ACCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTC ACCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGG GAGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTG GACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAG ACTTACAGGTTACCTCCGAAACTGA 270 L88T- ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCACCGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 271 L98T- ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATACCAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 272 N80Q-CAGE10 3N-AACK10 5T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGAACGAGACCCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 273 N80Q-CAGE10 4N-AACL10 6T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGAACAAGACCGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 274 N80Q-CAGE10 7N- AAC|E10 9T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGAACAAGACCGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 275 N80Q-CAGE10 9N- AAC|F111 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGAACGACACCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 276 N80Q-CAGE81 N-AAC|I83T -ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GAACACTACCGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA Til N80Q-CAGE85 N- AAC|L87 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTAACAACACCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 278 N80Q-CAGH97 N- AAC|K99 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CAACCTGACCACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 279 N80Q-CAGI83N AAC|E85 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTAACGTTACCAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 280 N80Q-CAGK10 5N-AAC|E10 7T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAACCTGACCAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 281 N80Q- CAG|K10 8N- AAC|D11 OT-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAACGAGACCTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 282 N80Q-CAGK99 N- AACV10 1T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAACACAACCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 283 N80Q-CAGL10 2N-AACE10 4T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCAACGAGACCAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 284 N80Q-CAGL10 6N-AAC|K10 8T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGAACGAGACCGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 285 N80Q-CAGL88 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCACCGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 286 N80Q-CAGL98 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATACCAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 287 N80Q-CAG|Q94 N- AAC|N96 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATAACATAAC CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 288 N80Q-CAGT10 ON- AACL10 2T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAAACGTCACCGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 289 N80Q-CAGT82 N- AAC|V84 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGAACATTACCGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 290 N80Q-CAGV10 1N-AAC|E10 3T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAAACCTGACCGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 291 N80Q-CAGV84 N- AAC|N86 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGCA GGAGACTATTAACGAGACCCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 292 Q72N-AAC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGAAACGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 293 Q72N- AAC|N80 Q-CAG ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGAAACGATTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 294 Q94N-AACN96 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATAACATAAC CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 295 R71N- AACD73 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAACCAAACCTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 296 R71N- AACD73 T- ACC|N80 Q-CAG ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAACCAAACCTCATCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 297 S74N-AAC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATAACTCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 298 S74N- AAC|N80 Q-CAG ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATAACTCTAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 299 S75N-AAC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCAAACAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 300 S75N- AACN80 Q-CAG ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCAAACAGCACTGGCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 301 S76N- AAC|G78 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAACACTACCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 302 S76N- AACG78 T- ACC|N80 Q-CAG ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAACACTACCTGGCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 303 T100N- AACL10 2T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAAACGTCACCGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 304 T58N- AACY60 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGAACATCACCGAGATGCTCC AGAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGA ATGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAA ACCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTC ACCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGG GAGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTG GACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAG ACTTACAGGTTACCTCCGAAACTGA 305 T77N- AACW79 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCAACGGCACCAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 306 T77N- AACW79 T- ACC|N80 Q-CAG ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCAACGGCACCCA GGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 307 T82N- AACV84 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGAACATTACCGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 308 V101N- AACE10 3T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAAACCTGACCGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 309 V84N- AACN86 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTAACGAGACCCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 310 W79N- AACE81 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCAACAA TACCACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 311 A135N- AACE13 7T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGAACAAGACCTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 312 E137N-AAC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGAACTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 313 F156N-AAC|N15 8T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACAACATTACCAGA CTTACAGGTTACCTCCGAAACTGA 314 G114N- AACL11 6T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGAACAAAACCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 315 G162N- AACL16 4T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAAACTACACCCGAAACTGA 316 H121N- AACK12 3T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGAACCTGACCAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 317 H131N- AACL13 3T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGAACTACACCAAGGCCAAGGAGTACAGTCACTGTGCCTG GACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAG ACTTACAGGTTACCTCCGAAACTGA 318 H140N-AACA14 2T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTAACTGTACCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 319 I145N- AACR14 7T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCAACGTGACCGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 320 K115N- AAC|M11 7T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAACCTCACCAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 321 K134N- AACK13 6T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAACGCCACCGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 322 K136N-AACY13 8T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAACGAGACCAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 323 L116N- AAC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAAAACATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 324 L122N-AACR12 4T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACAACAAAACCTATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 325 L130N- AACY13 2T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTAACCATACCCTGAAGGCCAAGGAGTACAGTCACTGTGCCTG GACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAG ACTTACAGGTTACCTCCGAAACTGA 326 L151N- AAC|N15 3T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCAACAGGACCTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 327 L160N-AACG16 2T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA AACACAACCTACCTCCGAAACTGA 328 L160T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA ACCACAGGTTACCTCCGAAACTGA 329 L164N- AACN16 6T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACAACCGAACCTGA 330 RI 13N-AAC|K11 5T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAACGGAACCCTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 331 R124N- AACY12 6T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAACTATACCGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 332 R128N- AACL13 OT-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AACATTACCCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 333 R147N- AACE14 9T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAACGTGACCATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 334 R159N- AAC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAAC CTTACAGGTTACCTCCGAAACTGA 335 S118N- AACL12 OT-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAACAGTACCCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 336 T112N- AAC|G11 4T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA ACAGGACCAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 337 T144N-AACV14 6T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG AACATAACCAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 338 T161N- AACY16 3T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTAACGGTACCCTCCGAAACTGA 339 V148N- AACI150 T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAAACGAAACCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 340 Y138N-AACH14 0T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGAACAGTACCTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 341 Y155T- ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTACCTTCATTAACAGA CTTACAGGTTACCTCCGAAACTGA 342 Y163N-AACR16 5T-ACC ATGAGCTACAACTTGCTTGGATTCCTACAAAGAAGCAGCAATTTTCAG TGTCAGAAGCTCCTGTGGCAACTTAATGGGAGGCTTGAATACTGCCTC AAGGACAGGATGAACTTTGACATCCCTGAGGAGATTAAGCAGCTGCA GCAGTTCCAGAAGGAGGACGCCGCATTGACCATCTATGAGATGCTCCA GAACATCTTTGCTATTTTCAGACAAGATTCATCTAGCACTGGCTGGAA TGAGACTATTGTTGAGAACCTCCTGGCTAATGTCTATCATCAGATAAA CCATCTGAAAACAGTCCTGGAGGAGAAGCTGGAGAAGGAGGACTTCA CCAGGGGAAAACTCATGAGCAGTCTGCACCTGAAAAGATATTATGGG AGGATTCTGCATTACCTGAAGGCCAAGGAGTACAGTCACTGTGCCTGG ACCATAGTGAGAGTGGAAATCCTAAGGAACTTTTACTTCATTAACAGA CTTACAGGTAACCTCACCAACTGA Compositions

[0097] Provided herein are compositions comprising an engineered interferon beta (IFN-beta) protein or a functional fragment thereof, comprising a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1, and a pharmaceutically acceptable excipient, diluent, or carrier. In some embodiments, the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208. In some embodiments, an engineered, non-naturally occurring protein having an amino acid substitution recited in TABLE 1, or a vector encoding the engineered protein having an amino acid substitution recited in TABLE 1, can be present in a composition. In some cases, the composition can be a pharmaceutical composition with a pharmaceutically acceptable excipient, diluent, or carrier. In some embodiments, an engineered, non-naturally occurring protein having an amino acid substitution recited in TABLE 2, or a vector encoding the engineered protein having an amino acid substitution recited in TABLE 2, can be present in a composition. In some cases, the composition can be a pharmaceutical composition with a pharmaceutically acceptable excipient, diluent, or carrier.

[0098] In some embodiments, a pharmaceutical composition can comprise an excipient, such as a buffering agent, a preservative, a stabilizer, a binder, a compaction agent, a lubricant, a chelator, a dispersion enhancer, a disintegration agent, a flavoring agent, a sweetener, a coloring agent. In some instances, a pharmaceutical composition can comprise a diluent such as water, glycerol, methanol, ethanol, and other similar biocompatible diluents. In some embodiments, the pharmaceutical composition can be in unit dose form.

[0099] In some embodiments, the pharmaceutical composition can be in the form of a tablet, a liquid, a syrup, an oral formulation, an intravenous formulation, an intranasal formulation, an ocular formulation, an otic formulation, a suppository, and any combination thereof. Methods of treatment

[00100] Also provided herein are methods for treating a disease or condition, the method comprising administering to a subject in need thereof a therapeutically effective dose of engineered interferon beta (IFN-beta) protein or a functional fragment thereof, comprising a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1. In some embodiments, the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208. In some embodiments, an engineered, non-naturally occurring protein having an amino acid substitution recited in TABLE 1 or TABLE 2, a vector encoding the non-naturally occurring protein having an amino acid substitution recited in TABLE 1, or a pharmaceutical composition comprising the non-naturally occurring protein or vector, can be administered to a subject to treat a disease or condition. For example, the non-naturally occurring protein having an amino acid substitution recited in TABLE 1, vector encoding the non-naturally occurring protein having an amino acid substitution recited in TABLE 1, or pharmaceutical composition comprising the non-naturally occurring protein or vector, can be used to treat an inflammatory condition. In some embodiments, the non-naturally occurring protein having an amino acid substitution recited in TABLE 1, vector encoding the non-naturally occurring protein having an amino acid substitution recited in TABLE 1, or pharmaceutical composition comprising the non-naturally occurring protein or vector, can be used to treat a cancer (e.g., a melanoma, a bladder cancer, a head or neck cancer, a kidney cancer, a liver cancer, a non-small cell lung cancer, a viral infection (e.g.. a hepatitis infection such as chronic Hepatitis B infection or a chronic Hepatitis C infection), a systemic lupus (e.g., erythematosus), or multiple sclerosis (e.g., relapse remitting MS, non-relapsing MS, clinically isolated syndrome, and secondary progressive forms).

[00101] In some embodiments, the non-naturally occurring protein having an amino acid substitution recited in TABLE 1. vector encoding the non-naturally occurring protein having an amino acid substitution recited in TABLE 1, or pharmaceutical composition comprising the non-naturally occurring protein or vector, can be administered to a subj ect by a route of administration selected from: inhalation, otic, buccal, conjunctival, dental, endocervical, endosinusial, endotracheal, enteral, epidural, extra-amniotic, extracorporeal, hemodialysis, infiltration, interstitial, intraabdominal, intraamniotic, intraarterial, intraarticular, intrabiliary, intrabronchial, intrabursal, intracardiac, intracartilaginous, intracaudal, intracavemous, intracavitary7, intracerebroventricular, intracistemal, intracorneal, intracoronal, intracoronary, intracorpous cavemaosum. intradermal, intradiscal, intraductal, intraduodenal, intradural, intraepidermal, intraesophageal, intragastric. intragingival, intrahippocampal. intraileal, intralesional, intraluminal, intralymphatic, intramedullary, intrameningeal, intramuscular, intraocular, intraovarian, intrapericardial, intraperitoneal, intrapleural, intraprostatic, intrapulmonary, intrasinal, intraspinal, intrasynovial, intratendinous, intratesticular. intrathoracic, intratubular, intratumor, intratympanic, intrauterine, intravascular, intravenous, intravenous bolus, intravenous drip, intravesical, intravitreal, iontophoresis, irrigation, laryngeal, nasal, nasogastric, ophthalmic, oral, oropharyngeal, parenteral, percutaneous, periarticular, peridural, perineural, periodontal, rectal, retrobulbar, subarachnoid, subconjunctival, subcutaneous, sublingual, submucosal, topical, transdermal, transmucosal, transplacental, transtracheal, transtympanic, ureteral, urethral, vaginal, infraorbital, intraparenchymal, intrathecal, intraventricular, stereotactic, or any combination thereof.

[00102] In some embodiments, the non-naturally occurring protein having an amino acid substitution recited in TABLE 1. vector encoding the non-naturally occurring protein having an amino acid substitution recited in TABLE 1. or pharmaceutical composition comprising the non-naturally occurring protein or vector, can be co-administered along with a second therapeutic. In some embodiments, the second therapeutic can be an antibody, a peptide, an antibody drug conjugate, or any combination thereof. In some cases, the second therapeutic can be an immunotherapy agent such as a checkpoint inhibitor. In some embodiments, the checkpoint inhibitor can be a PD-1 inhibitor such as nivolumab or pembrolizumab, a CTLA-4 inhibitor such as ipilimumab, or a PD-L1 inhibitor such as atezolizumab, avelumab, or durvalumab.

[00103] In some embodiments, the non-naturally occurring protein having an amino acid substitution recited in TABLE 1. vector encoding the non-naturally occurring protein having an amino acid substitution recited in TABLE 1, or pharmaceutical composition comprising the non-naturally occurring protein or vector, can be administered 1, 2, 3, or 4 times a day; 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 15, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 times a week; or 1. 2, 3, 4, 5, 6, 7, 8. 9, 10, 11, 12, 13. 14. 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29. 30, 31, 32, 33, 34, 35, 36, 37, 38, 39. 40. 41. 42. 43. 44. 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55. 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, or 90 times a month. In some embodiments, the non-naturally occurring protein having an amino acid substitution recited in TABLE 1, vector encoding the non-naturally occurring protein having an amino acid substitution recited in TABLE 1. or pharmaceutical composition comprising the non-naturally occurring protein or vector, can be administered every day, every other day, once a week, or once a month.

[00104] In some embodiments, the non-naturally occurring protein having an amino acid substitution recited in TABLE 1. vector encoding the non-naturally occurring protein having an amino acid substitution recited in TABLE 1. or pharmaceutical composition comprising the non-naturally occurring protein or vector, can independently be administered at a dose selected from the group consisting of about: 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 0.6 mg / kg, 0.7 mg / kg, 0.8 mg / kg, 0.9 mg / kg, 1 mg / kg, 2 mg / kg, 3 mg / kg. 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, and 10 mg / kg; each with respect to a body weight of the subject.

[00105] In some embodiments, the non-naturally occurring protein having an amino acid substitution recited in TABLE 1, vector encoding the non-naturally occurring protein having an amino acid substitution recited in TABLE 1. or pharmaceutical composition comprising the non-naturally occurring protein or vector, can independently be administered at a dose of about: 10 ng, 11 ng, 12 ng, 13 ng, 14 ng, 15 ng, 16 ng, 17 ng, 18 ng, 19 ng, 20 ng, 21 ng, 22 ng, 23 ng, 24 ng, 25 ng, 26 ng, 27 ng, 28 ng, 29 ng, 30 ng, 31 ng, 32 ng, 33 ng, 34 ng, 35 ng, 36 ng, 37 ng, 38 ng, 39 ng, 40 ng, 41 ng, 42 ng, 43 ng, 44 ng, 45 ng, 46 ng. 47 ng, 48 ng, 49 ng, 50 ng, 51 ng, 52 ng, 53 ng, 54 ng. 55 ng, 56 ng. 57 ng, 58 ng. 59 ng, 60 ng. 61 ng, 62 ng, 63 ng, 64 ng, 65 ng. 66 ng, 67 ng, 68 ng, 69 ng, 70 ng, 71 ng, 72 ng, 73 ng, 74 ng, 75 ng, 76 ng, 77 ng, 78 ng, 79 ng, 80 ng, 81 ng, 82 ng, 83 ng, 84 ng, 85 ng, 86 ng, 87 ng, 88 ng, 89 ng, 90 ng, 91 ng, 92 ng, 93 ng, 94 ng, 95 ng, 96 ng, 97 ng, 98 ng, 99 ng, 100 ng, 110 ng, 120 ng, 130 ng, 140 ng, 150 ng, 160 ng, 170 ng. 180 ng, 190 ng, 200 ng, 210 ng, 220 ng. 230 ng, 240 ng, 250 ng, 260 ng, 270 ng. 280 ng. 290 ng, 300 ng, 310 ng, 320 ng, 330 ng, 340 ng, 350 ng, 360 ng, 370 ng, 380 ng, 390 ng, 400 ng, 410 ng, 420 ng, 430 ng, 440 ng, 450 ng, 460 ng, 470 ng, 480 ng, 490 ng, 500 ng, 510 ng, 520 ng, 530 ng, 540 ng, 550 ng, 560 ng, 570 ng, 580 ng, 590 ng, 600 ng, 610 ng, 620 ng, 630 ng, 640 ng, 650 ng, 660 ng, 670 ng, 680 ng, 690 ng, 700 ng, 710 ng, 720 ng, 730 ng, 740 ng, 750 ng, 760 ng. 770 ng, 780 ng, 790 ng, 800 ng, 810 ng, 820 ng, 830 ng, 840 ng, 850 ng, 860 ng, 870 ng, 880 ng, 890 ng, 900 ng, 910 ng, 920 ng, 930 ng, 940 ng, 950 ng, 960 ng, 970 ng, 980 ng, 990 ng, 1 jig, 1.1 jig, 1.2 jig, 1.3 jig, 1.4 jig, 1.5 jig, 1.6 jig, 1.7 jig, 1.8 jig, 1.9 jig, 2 jig, 2.1 jig, 2.2 jig, 2.3 jig, 2.4 jig, 2.5 jig, 2.6 jig, 2.7 jig, 2.8 jig, 2.9 jig, 3 jig, 3.1 jig, 3.2 jig, 3.3 jig, 3.4 jig, 3.5 jig, 3.6 jig, 3.7 jig, 3.8 jig, 3.9 jig, 4 jig, 4.1 jig, 4.2 jig, 4.3 jig, 4.4 jig, 4.5 jig, 4.6 jig, 4.7 jig, 4.8 jig, 4.9 jig, 5 jig, 5.1 jig, 5.2 jig, 5.3 jig, 5.4 jig, 5.5 jig, 5.6 jig, 5.7 jig, 5.8 jig, 5.9 jig, 6 jig, 6.1 jig, 6.2 jig, 6.3 jig, 6.4 jig, 6.5 jig, 6.6 jig, 6.7 jig, 6.8 jig, 6.9 jig, 7 jig, 7.1 jig, 7.2 jig, 7.3 jig, 7.4 jig, 7.5 jig, 7.6 jig, 7.7 jig, 7.8 jig, 7.9 jig, 8 jig, 8.1 jig, 8.2 jig, 8.3 jig, 8.4 jig, 8.5 jig, 8.6 jig, 8.7 jig, 8.8 jig, 8.9 jig, 9 jig, 9.1 jig, 9.2 jig, 9.3 jig, 9.4 pig, 9.5 pig, 9.6 gg, 9.7 jig, 9.8 jig, 9.9 jig, 10 jig. 11 jig, 12 jig, 13 jig, 14 jig, 15 jig, 16 gg, 17 gg, 18 gg, 19 gg, 20 gg, 21 gg, 22 gg, 23 gg, 24 gg, 25 jig, 26 gg, 27 gg, 28 jig, 29 gg, 30 jig, 31 jig, 32 gg, 33 jig, 34 gg, 35 jig, 36 jig, 37 gg, 38 jig, 39 gg, 40 jig, 41 jig, 42 jig, 43 gg, 44 gg, 45 gg, 46 gg, 47 gg, 48 gg, 49 gg, 50 jig, 51 gg, 52 gg, 53 jig, 54 jig. 55 jig, 56 gg, 57 gg, 58 gg, 59 gg, 60 gg, 61 gg, 62 gg, 63 gg, 64 gg, 65 gg, 66 jig, 67 jig. 68 jig. 69 gg, 70 gg, 71 gg, 72 gg, 73 gg, 74 gg, 75 gg, 76 gg, 77 gg, 78 gg, 79 jig, 80 jig, 81 jig, 82 jig, 83 jig, 84 gg, 85 gg, 86 gg, 87 gg, 88 gg, 89 gg, 90 gg, 91 gg, 92 gg, 93 jig, 94 jig, 95 jig, 96 gg, 97 gg, 98 gg, 99 gg, 100 gg, 110 gg, 120 gg, 130 gg, 140 gg, 150 jig, 160 jig. 170 jig, 180 jig, 190 jig, 200 jig, 210 gg, 220 gg, 230 gg, 240 gg, 250 gg, 260 gg, 270 jig. 280 jig, 290 gg, 300 gg, 310 gg, 320 gg, 330 gg, 340 gg. 350 gg, 360 gg, 370 gg, 380 jig, 390 jig, 400 jig, 410 jig, 420 gg, 430 gg, 440 gg, 450 gg, 460 gg, 470 gg, 480 gg, 490 gg, 500 jig, 510 jig, 520 jig, 530 gg, 540 gg, 550 gg, 560 gg, 570 gg, 580 gg, 590 gg, 600 gg, 610 jig, 620 jig, 630 jig, 640 gg, 650 gg, 660 gg, 670 gg, 680 gg, 690 gg, 700 gg, 710 gg, 720 gg, 730 jig. 740 jig, 750 gg, 760 gg, 770 gg, 780 gg, 790 gg, 800 gg. 810 gg, 820 gg, 830 gg. 840 jig, 850 jig, 860 jig, 870 gg, 880 gg, 890 gg, 900 gg, 910 gg, 920 gg, 930 gg, 940 gg, 950 gg, 960 jig, 970 jig, 980 gg, 990 gg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3 mg. 3.1 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.5 mg, 3.6 mg, 3.7 mg, 3.8 mg, 3.9 mg. 4 mg, 4.1 mg, 4.2 mg. 4.3 mg, 4.4 mg, 4.5 mg, 4.6 mg. 4.7 mg, 4.8 mg, 4.9 mg, 5 mg. 5.1 mg. 5.2 mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg, 5.8 mg, 5.9 mg, 6 mg, 6.1 mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7 mg, 7.1 mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg, 7.6 mg, 7.7 mg, 7.8 mg, 7.9 mg, 8 mg, 8.1 mg, 8.2 mg, 8.3 mg, 8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg. 8.8 mg, 8.9 mg, 9 mg. 9.1 mg, 9.2 mg, 9.3 mg, 9.4 mg. 9.5 mg, 9.6 mg, 9.7 mg, 9.8 mg, 9.9 mg. 10 mg. 11 mg. 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 mg, 23 mg, 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg. 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg. 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg. or 100 mg. EMBODIMENTS

[00106] Embodiment 1. A non-naturally occurring protein, comprising an amino acid sequence having an amino acid substitution with respect to a wildtype interferon beta (IFN-beta) protein with the amino acid sequence of SEQ ID NO: 1. wherein the amino acid substitution is selected from the amino acid substitutions in TABLE 1.

[00107] Embodiment 2. The non-naturally occurring protein of embodiment 1, wherein: a. the amino acid substitution results in a conversion of a motif in the amino acid sequence of SEQ ID NO: 1 that is not a glycosylation motif into a glycosylation motif, thereby introducing a glycosylation motif into the non-naturally occurring protein; b. the amino acid substitution results in a conversion of a glycosylation motif in the amino acid sequence of SEQ ID NO: 1 into a motif that is not a glycosylation motif, thereby eliminating a glycosylation motif from the non-naturally occurring protein; or c. a combination of (a) and (b).

[00108] Embodiment 3. The non-naturally occurring protein of embodiment 2, wherein the amino acid substitution results in the conversion of 1, 2, 3, 4, or more motif(s) in the amino acid sequence of SEQ ID NO: 1 that are not glycosylation motifs into 1, 2, 3, 4, or more motif(s) that are glycosylation motif(s), thereby introducing 1, 2, 3. 4, or more glycosylation motif(s) into the non-naturally occurring protein.

[00109] Embodiment 4. The non-naturally occurring protein of embodiment 2, wherein the amino acid substitution results in the conversion of 1, 2, 3, 4, or more glycosylation motif(s) in the amino acid sequence of SEQ ID NO: 1 into 1, 2, 3, 4, or more motif(s) that are not glycosylation motifs, thereby eliminating 1, 2. 3, 4, or more glycosylation motif(s) into the non-naturally occurring protein.

[00110] Embodiment 5. The non-naturally occurring protein of any one of embodiments 1-4, wherein the glycosylation motif has a consensus sequence of N-X-S / T, wherein N is an Asparagine that is N-glycosylated at the amide bond in the side chain of the Asparagine, X is any amino acid except for proline, S is a serine, and T is a threonine.

[00111] Embodiment 6. The non-naturally occurring protein of embodiment 5, wherein the amino acid substitution in the non-naturally occurring protein is selected from the amino acid substitutions in TABLE 2.

[00112] Embodiment 7. The non-naturally occurring protein of embodiment 5, wherein an asparagine, a serine, a threonine, or any combination thereof, is substituted.

[00113] Embodiment 8. The non-naturally occurring protein of embodiment 1, wherein the amino acid substitution results in a conversion of a motif in the amino acid sequence of SEQ ID NO: 1 that is not a site for a post-translational modification into a post-translational modification consensus sequence motif, thereby introducing a site for a post translational modification into the non-naturally occurring protein.

[00114] Embodiment 9. The non-naturally occurring protein of embodiment 8, wherein the post-translational modification consensus sequence motif is selected from the amino acid sequences in TABLE 5.

[00115] Embodiment 10. The non-naturally occurring protein of embodiment 9, wherein the amino acid substitution results in the addition of a post-translational modification on the non-naturally occurring protein at the site of the post-translational modification consensus sequence motif that is not present in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1.

[00116] Embodiment 11. The non-naturally occurring protein of embodiment 10, wherein the post-translational modification is selected from the group consisting of: glycosylation, phosphorylation, lipidation, glycation, proline isomerization, methylation, acetylation, hydroxylation sites, and proteolysis.

[00117] Embodiment 12. The non-naturally occurring protein of any one of embodiments 1-11, wherein the non-naturally occurring protein is glycosylated.

[00118] Embodiment 13. The non-naturally occurring protein of any one of embodiments 1-11, wherein the non-naturally occurring protein is aglycosylated.

[00119] Embodiment 14. The non-naturally occurring protein of any one of embodiments 1-13, wherein the non-naturally occurring protein binds to an interferon-a / p receptor-l(IFNARl) or interferon-a / p receptor-2 (IFNAR2) heterodimer subunits at a level greater than or equal to the wildtype IFN-beta protein as measured by enzyme-linked immunosorbent assay (ELISA) or by measuring signaling activity of an Interferon Stimulated Response Element (ISRE) Green fluorescent protein (GFP) reporter in alive human cell in vitro assay.

[00120] Embodiment 15. The non-naturally occurring protein of any one of embodiments 1-14, wherein the non-naturally occurring protein is an agonist of IFNAR1 or IFNAR2 subunits.

[00121] Embodiment 16. The non-naturally occurring protein of embodiment 15, wherein the non-naturally occurring protein binds and activates IFNAR1 or IFNAR2 at a level greater than the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by an ISRE-GFP reporter assay or an ISRE luciferase reporter assay.

[00122] Embodiment 17. The non-naturally occurring protein of any one of embodiments 1-13, wherein the non-naturally occurring protein reduces an extracellular level of a pro-inflammatory cytokine, as compared to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by ELISA.

[00123] Embodiment 18. The non-naturally occurring protein of embodiment 17, wherein the pro-inflammatory cytokine is a chemokine, an interferon (IFN), an interleukin (IL), a lymphokine, a tumor necrosis factor (TNF), or any combination thereof.

[00124] Embodiment 19. The non-naturally occurring protein of embodiment 18, wherein the pro-inflammatory cytokine is IL-Ip, IL-6, IL-8, IL-9, IL-12, IL-15, IL-17, IL-18. IFN-y, TNF-a, TNF-P, or any combination thereof.

[00125] Embodiment 20. The non-naturally occurring protein of any one of embodiments 1-13, wherein the non-naturally occurring protein increases an extracellular level of an antiinflammatory cytokine, as compared to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by ELISA.

[00126] Embodiment 21. The non-naturally occurring protein of embodiment 20, wherein the anti-inflammatory cytokine is a chemokine, an IFN, an IL, a lymphokine, a TNF, a Transforming growth factor (TGF), or any combination thereof.

[00127] Embodiment 22. The non-naturally occurring protein of embodiment 21, wherein the anti-inflammatory cytokine is IL-4, IL-10, IL-11, IL-13, IFN-a, TGF-P, or any combination thereof.

[00128] Embodiment 23. The non-naturally occurring protein of any one of embodiments 1-13, wherein the non-naturally occurring protein binds to IFNAR1 or IFNAR2 subunits at a level less than the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by ELISA.

[00129] Embodiment 24. The non-naturally occurring protein of any one of embodiments 1-14, wherein the non-naturally occurring protein is an antagonist of IFNAR1 or IFNAR2 subunits.

[00130] Embodiment 25. The non-naturally occurring protein of embodiment 24, wherein the non-naturally occurring protein binds and inhibits the activity of the IFNAR1 or IFNAR2 at a level greater than wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by an ISRE GFP reporter assay or an ISRE luciferase reporter assay.

[00131] Embodiment 26. The non-naturally occurring protein of any one of embodiments 1-25, wherein the non-naturally occurring protein is a recombinant IFN-beta or a portion thereof.

[00132] Embodiment 27. The non-naturally occurring protein of any one of embodiments 1-26, wherein the non-naturally occurring protein is glycosylated, carboxylated, hydroxylated, sulfated, phosphorylated, albuminated, conjugated to a polyethylene glycol (PEG) moiety, or any combination thereof.

[00133] Embodiment 28. The non-naturally occurring protein of embodiment 1-27, wherein a glycosylation site is introduced in the non-naturally occurring protein that results in diminished binding of an antibody to the non-naturally occurring protein relative to an amount of binding of the antibody to the wildtype IFN-beta protein.

[00134] Embodiment 29. The non-naturally occurring protein of any one of embodiments 1-28, wherein the non-naturally occurring beta protein has reduced immunogenicity' when administered to a subject, as compared to administering the wildtype IFN-beta with the amino acid sequence ofSEQIDNO: 1.

[00135] Embodiment 30. The non-naturally occurring protein of embodiment 28, wherein the reduced immunogenicity comprises a reduced level of anti-drug antibody in the subject when the non-naturally occurring protein is administered to the subject, as compared a level of anti-drug antibody when the wildtype IFN-beta with the amino acid sequence of SEQ ID NO: 1 is administered, as measured by ELISA on a sample obtained from the subject.

[00136] Embodiment 31. The non-naturally occurring protein of any one of embodiments 1-30, wherein the non-naturally occurring beta protein, when administered to a subject in need thereof, reduces a level of inflammation in a subject, relative to a level of inflammation in the subject prior to the administering.

[00137] Embodiment 32. The non-naturally occurring protein of embodiment 31, wherein the reduced level of inflammation is determined by analyzing levels of an autoantibody, a C reactive protein (CRP), a proteolytic enzy me, an inflammatory mediator, a marker of ongoing inflammation, or any combination thereof, as measured by ELISA.

[00138] Embodiment 33. The non-naturally occurring protein of embodiment 31, wherein the reduced level of inflammation in the subject results in reduced serum levels of C reactive protein as measured by high-sensitivity C-reactive protein (hs-CRP) test.

[00139] Embodiment 34. The non-naturally occurring protein of embodiment 33, wherein the reduced serum levels of C reactive protein comprises a plasma concentration in the subject of from about 0.1 mg / dL to about 2.9 mg / dL.

[00140] Embodiment 35. The non-naturally occurring protein of embodiment 34, wherein the administering to the subject results in reduced blood levels of a pro-inflammatory cytokine as compared to the blood levels of one or more pro-inflammatory cytokines before the administering, as measured by ELISA.

[00141] Embodiment 36. A method of making a non-naturally occurring protein in a cell, comprising introducing into the cell a vector encoding the non-naturally occurring protein of any one of embodiments 1-35, wherein the introducing results in expression of the non-naturally occurring protein in the cell.

[00142] Embodiment 37. The method of embodiment 36, wherein the non-naturally occurring protein is isolated from the cell.

[00143] Embodiment 38. The method of embodiment 36, wherein the vector comprises a mammalian expression plasmid.

[00144] Embodiment 39. The method of embodiment 36, wherein the non-naturally occurring protein comprises a tag.

[00145] Embodiment 40. The method of embodiment 39, wherein the tag is CBP, FLAG, GST. Myc, poly-His, or V5.

[00146] Embodiment 41. The method of embodiment 39, wherein the tag comprises a cleavable tag or a non-cleavable tag.

[00147] Embodiment 42. A polynucleotide encoding the non-naturally occurring protein of any one of embodiments 1-35.

[00148] Embodiment 43. A pharmaceutical composition in unit dose form comprising: a. the non-naturally occurring protein of any one of embodiments 1-35, a vector encoding the non-naturally occurring protein of any one of embodiments 1-35, or the polynucleotide of embodiment 42; and b. a pharmaceutically acceptable excipient, carrier, or diluent.

[00149] Embodiment 44. The pharmaceutical composition of embodiment 43, wherein the pharmaceutical composition is encapsulated.

[00150] Embodiment 45. The pharmaceutical composition of embodiment 43, wherein the pharmaceutical composition is in the form of a liquid.

[00151] Embodiment 46. The pharmaceutical composition of any one of embodiments 43-45, wherein the pharmaceutical composition is formulated for local, systemic, or topical administration.

[00152] Embodiment 47. The pharmaceutical composition of any one of embodiments 43-46, wherein the pharmaceutical composition is formulated for oral, nasal, pulmonary, buccal, transdermal, subcutaneous, intraduodenal, enteral, parental, intravenous, or intramuscular administration.

[00153] Embodiment 48. The pharmaceutical composition of any one of embodiments 43-47, wherein the pharmaceutical composition is formulated for controlled-release.

[00154] Embodiment 49. The pharmaceutical composition of any one of embodiments 43-48, wherein the pharmaceutical composition is formulated for single-dosage administration.

[00155] Embodiment 50. A method of treating a disease in a subject, the method comprising: administering to the subject a therapeutically effective amount of the non-naturally occurring protein of any one of embodiments 1-35, the polynucleotide of embodiment 42, or the pharmaceutical composition of any one of embodiments 43-49, thereby treating the subject.

[00156] Embodiment 51. The method of embodiment 50, wherein the disease is an infection.

[00157] Embodiment 52. The method of embodiment 51, wherein the infection is a viral infection.

[00158] Embodiment 53. The method of embodiment 52, wherein the viral infection is chronic Hepatitis B infection, or a chronic Hepatitis C infection.

[00159] Embodiment 54. The method of embodiment 50, wherein the disease is a demyelinating disorder.

[00160] Embodiment 55. The method of embodiment 54, wherein the demyelinating disorder is selected from the group consisting of Multiple Sclerosis (MS), Acute Disseminated Encephalomyelitis (ADEM), Acute Hemorrhagic Leucoencephalitis (AHLE), Balo’s disease (Concentric Sclerosis), Charcot-Marie-Tooth disease (CMT), Guillain-Barre Syndrome (GBS), HTLV-I Associated Myelopathy (HAM), and Neuromyelitis Optica (Devic’s Disease).

[00161] Embodiment 56. The method of embodiment 55, wherein the MS is relapsing remitting MS, non-relapsing MS, clinically isolated syndrome, and secondary' progressive forms.

[00162] Embodiment 57. The method of any one of embodiments 50-56, wherein the subject is a human subject.

[00163] Embodiment 58. The method of any one of embodiments 50-57, wherein the administering is by parenchymal injection, intra-thecal injection, intra-ventricular injection, intra-cistemal injection, intratumoral injection, subcutaneous injection, intraperitoneal injection, a surgical route, or any combination thereof.

[00164] Embodiment 59. The method of embodiment 50, wherein the administering occurs from about once a day, about twice a day, about once a week, about twice a week, about once every' two weeks, about once a month, about once every' 3 months, about once every' 6 months, about once every 9 months, to about once a year.

[00165] Embodiment 60. The method of embodiment 50, wherein the administering is at a dose selected from the group consisting of about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 700 mg, about 800 mg, about 900 mg. about 1000 mg, about 1200 mg, about 1500 mg, or about 2000 mg.

[00166] Embodiment 61. The method of embodiment 50, wherein the administering is at a dose selected from the group consisting of 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, and 10 mg / kg.

[00167] Embodiment 62. The method of embodiment 50, wherein the administering reduces a volume of a tumor in the subject.

[00168] Embodiment 63. The method of embodiment 50, wherein the administering inhibits or diminishes the grow th a tumor in the subject.

[00169] Embodiment 64. The method of embodiment 50, wherein the method further comprises administering a second therapeutic.

[00170] Embodiment 65. The method of embodiment 64, wherein the second therapeutic is an antibody, a peptide, an antibody drug conjugate, a CAR-T, a cancer vaccine, or any combination thereof.

[00171] Embodiment 66. The method of embodiment 50, wherein the method further comprises administering an immunotherapy, and wherein the immunotherapy comprises a checkpoint inhibitor.

[00172] Embodiment 67. The method of embodiment 66, wherein the checkpoint inhibitor is a PD-1, aPD-Ll, CTLA-4 inhibitor.

[00173] Embodiment 68. The method of embodiment 50, wherein the administering induces activation of an IFN-stimulated response element (ISRE) or a Gamma interferon activation site (GAS) element, wherein the activation of the ISRE or GAS induces an interferon-like activity as measured by a luciferase reporter assay.

[00174] Embodiment 69. A kit comprising the non-naturally occurring protein of any one of embodiments 1-35, the polynucleotide of embodiment 42, or the pharmaceutical composition of any one of embodiments 43-49, and a container. EXAMPLES Example 1 - Generation of a plasmid-based Gene Mutation Library

[00175] A library' of non-naturally occurring Interferon beta variants was generated. The library was a plasmid library designed for the generation of a lentiviral library and contains the cDNA for human Interferon beta having the sequence of SEQ ID NO: 1 with variants as described in TABLE 1, and TABLE 2. (FIG. 1). A clonal reporter cell line was established that contains a stably integrated ZsGreenl-DR fluorescent protein downstream of an interferon-responsive promoter containing 3xISREs (Interferon Stimulated Response Elements). (FIG. 2) The reporter cell line was then transduced so that it contains a single integrated copy of a non-naturally occurring protein as described herein downstream of a Dox-inducible promoter. The non-naturally occurring protein as described herein consists of an N-terminal fusion of a Type II Transmembrane Domain (Type II TMD) and GGGGS linker. Upon addition of doxycy cline, the Tet Transactivator binds doxycycline and changes conformation to allow for binding to the Tet response elements in the Dox-inducible promoter leading to the induced expression of the non-naturally occurring protein as described herein. The non-naturally occurring protein as described herein is then translocated to the plasma membrane where it is tethered to the membrane with the Type II TMD. On the cell surface, non-naturally occurring protein as described herein interacts with the Type IIFN receptor (consisting of IFNAR1 / 2 receptor subunits) to induce the interferon-stimulated signaling pathway. Phosphorylated STAT1 / 2 translocates to the nucleus, binds to the ISRE promoter upstream of GFP leading to induction of GFP expression. Cells can be sorted based on fluorescence as a marker for biological activity related to the interferon signaling pathway. (FIG. 2). The non-naturally occurring protein as described herein was saturation mutagenized in order to produce amino acid substitutions at each position thereby introducing or eliminating adding mutations in a consensus sequence as disclosed herein for N-linked glycosylation at the asparagine (N), serine (S), or threonine (T) residues of the consensus sequence motif of Nx[S / T], IFN-beta variants in the assay are designed so that N-linked glycosylation sites are added at each amino acid position within the protein coding sequence. Each clone generated contained a Unique Molecular Identifier (UMI) region which was used to quantify7 and associate the UMI in each sorted population with the corresponding gene mutation. This information is used to create MEGA-Maps associating gene mutations with biological activity. Example 2 - High-throughput screening of non-naturally occurring protein

[00176] The resulting IFN-beta variant lentiviral library produced in Example 1 was transduced into a cell population of about 30 million cells, stimulated with Doxycycline, and plated at low density. Plating densities were titrated to minimize paracrine (intercellular) signaling. Cells stably expressing IFN-P plus a red fluorescent protein (mScarlet) w ere co-cultured with cells expressing endogenous IFNAR1 / 2 and an ectopic ISRE reporter driving expression of a green fluorescent protein (ZsGreenl-DR). In this assay, only cells that undergo paracrine signaling become positive for ZsGreenl. This density was used for autocrine assays. Cells were harvested, fixed, and partitioned into bins by Flow Sorting based on zsGreen fluorescence (as a marker of biological activity') into four bins: Bin 1: Low fluorescence = Low biological activity Bin 2: Mid-Low fluorescence = Mid-Low biological activity Bin 3: Mid-High fluorescence = Mid-High biological activity Bin 4: High fluorescence = High biological activity

[00177] This population of barcodes was compared to the set of barcodes for wildtype IFN-beta (control) and used to identify variants that have binding activity similar to that of WT, which were separated from variants that have significantly lower activity than wildtype IFN-beta (FIG. 3, TABLE 7). A portion of the variants are aglycosylated, single-glycosylated, or doubleglycosylated, where the double-glycosylated variants include a de novo glycosylation site in combination with the native glycosylation site left intact. A wide dynamic range of fluorescence in HEK-293 ISRE reporter cells was observed when comparing control (left) to cells stimulated by the non-naturally occurring proteins described herein (right) by flow cytometry (FIG. 4). TABLE 7. Exemplary Engineered, Non-Naturally Occurring protein classification Non-Naturally Occurring protein class Number Reduced activity compared to wildtype IFN-beta protein 241 Wildtype IFN-beta protein 887

[00178] This high-throughput screening assay assessed the first 109 amino acids of the wildtype IFN-beta protein having the amino acid sequence of SEQ ID NO: 1 which produced approximately 887 mutants, including glycosylation mutants that had signaling activity equivalent to the wildtype IFN-beta protein (TABLE 7). Exemplary non-naturally occurring proteins comprised a glycosylation site in an immune epitope without any overlap to amino acid residues associated with binding to the 1FNAR1 / 2 receptor subunits. From this screen, novel non-naturally interferon beta variants containing de novo glycosylation site(s) or aglycosylation site(s) as described herein were discovered to have substantially similar, decreased signaling activity, or increased signaling activity to wildtype IFN-beta. Example 3 - Comparison between variants functional elements and structure

[00179] Based on the foregoing examples, several non-naturally occurring proteins were engineered to have receptor stimulation efficacy similar to that of wildtype IFN-beta. These novel non-naturally occurring proteins were designed to have increased serum half-life, increased solubility and reduced immunogenicity. The major immuno epitopes that patients develop during treatment with wildtype IFN-beta were amino acids 1-12, 151-162, and 121-132, with the first two being favored for neutralizing antibodies and amino acids 1-12 being the preferred epitope. When plotted on the 3D structure of IFNAR2 (PDB: 3SE3). epitopes 1-12 were directly juxtaposed to epitopes 151-162 and 121-132, with 121-132 on the opposing face of the protein. However, none of the epitopes were near the native N-linked glycosylation site of wildtype IFN-BETA at amino acid N80.

[00180] A model of the structure of a human IFN-beta paralog, IFN alpha 2 complexed with IFNAR receptor (PDB: 3SE3) was used to assess the contacts of the non-naturally occurring protein as described herein with the IFNAR receptor. The model was consistent with the contacts observed in the structure of the mouse IFNARl / IFN-beta complex (PDB: 3WCY). IFNAR2 binds to IFN-beta making contacts with amino acids 19, 22, 26, 26, 30, 33, 151, 155, 156, and 159. Then, IFNAR2 was recruited and contacts IFN-beta at amino acids 65, 67. 68. 88, 92, 119, 123, 126. and 127. IFNAR1 makes no contacts with IFNAR2. rather both directly bind IFN-beta.

[00181] The non-naturally occurring proteins containing the amino acid substitutions in TABLE 2 were mapped on the X-ray structure and a new IFNAR1 binding contact was discovered to be essential for signaling. The crystal structure of IFN-beta shows an asymmetric dimer with a patch of residues forming intermolecular contacts. IFN-beta was a hydrophobic protein and was prone to aggregation and may form dimers, although the relevance of these dimers to signaling was not well understood. The mutations in the non-naturally occurring proteins having increased biological activity' were found to be juxtaposed to the dimerization site. Moreover, two clusters of non-naturally occurring variants were immediately juxtaposed to the dimerization interface, thus these mutations can affect the dimerization or oligomerization. Notably these patches were not in immuno epitopes or IFNAR 1 / 2 receptor binding sites. Additionally, protein aggregation can generate an immune response and induction of antidrug antibodies, however, introduction of de novo glycosylation sites as described herein over the surface of the non-naturally occurring protein can block hydrophobic surfaces resulting in reduced protein aggregation, increased solubility and reduced immunogenicity when administered to a human subject. Example 4 - Purification of HIS-tagged non-naturally occurring proteins

[00182] TurboCHO cells is transfected with a mammalian expression plasmid to express HIS-tagged non-naturally occurring proteins (FIG. 5). A secretory signal is fused on the N-terminus of the 6xHIS tag to facilitate secretion of the protein into supernatant during cell culture. The secretory signal gets cleaved from the HIS-tagged IFN protein during protein trafficking. The supernatant from transfected cells is loaded onto aNickle-NTA (Ni-NTA) affinity column, which binds the HIS residue of the HIS-tagged non-naturally occurring protein. Wash buffers is loaded into the column to wash away unbound material, and elution buffer is used to release the purified HIS-tagged IFN protein from the column. The purified non-naturally occurring protein as described is checked for quality control using SDS-PAGE gels (to confirm purity' and size; FIGs. 15A-15N) and Bradford assay (to measure concentration). The yield and purity of each protein is listed in TABLE 8. TABLE 8. Purification results. Protein SEQ ID NO: Class Purification Yield (mg) from 30 mL Purity (%) Wildtype 1 WT 0.242 80 Wildtype 1 WT 0.425 85 K105N E107T 2 WT 0.48 85 M117N 3 WT 0.396 90 E107N E109T 4 WT 0.525 70 S119N H121T 5 GOF-like 0.494 85 Y92N Q94T 137 Reduced Function 0.615 95 RUN 6 WT 0.585 95 N80Q K99N V101T 75 GOF-like 0.010 NA Q72N 85 WT 0.225 35 A89M 136 GOF-like 0.570 95 L151N N153T 119 WT 0.009 NA RUN K105N E107T 343 Derivative 0.648 95 RUN M117N 344 Derivative 0.774 95 Example 5 - Screening of purified HIS-tagged leads with cell-based reporter assay

[00183] HEK-293 cells stably expressing ZsGreenl-DR fluorescent protein downstream of an interferon-responsive promoter containing 3xISREs (Interferon-Stimulated Response Elements) were plated in a 96-well plate and allowed to attach overnight (FIG. 6A). Once attached, cells were treated with serially-diluted purified engineered, non-naturally occurring proteins as described herein (EXAMPLE 4) (in triplicate; n=3) at varying concentrations covering the range of the dose curve. Fluorescence of the treated cells was measured on a plate reader 24 hours posttreatment and EC50 values for ISRE reporter activity was determined for each IFN lead. TABLE 9 provides the amino acid sequence for the two control sequences used in this example. TABLE 9. Control IFN-beta sequences SEQ ID NO MUTANT Amino Acid Sequence 343 RUN, K105N, E107T MSYNLLGFLQNSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPE EIKQLQQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVEN LLANVYHQINHLKTVLEENLTKEDFTRGKLMSSLHLKRYYGRI LHYLKAKEYSHCAWTIVRVEILRNFYFINRLTGYLRN 344 RUN, M117N MSYNLLGFLQNSSNFQCQKLLWQLNGRLEYCLKDRMNFDIPE EIKQLQQFQKEDAALTIYEMLQNIFAIFRQDSSSTGWNETIVEN LLANVYHQINHLKTVLEEKLEKEDFTRGKLNSSLHLKRYYGRI LHYLKAKEYSHCAWTIVRVEILRNFYFINRLTGYLRN-

[00184] Dose response curve output from the 96-well ISRE reporter assay with a calculated EC50 of 0.1161 ng / mL for wildtype IFN-beta protein (FIG. 6B; FIGs. 16A-16L). EC50 for cellbased reporter assay was measured in ng / m, and p-values were False Discovery Rate (FDR) adjusted (FIGs. 16A-16L). The HIS-tagged IFN-beta protein with biosimilar activity was also verified by flow cytometry in cells stimulated with purified engineered IFN-beta proteins or WT proteins (FIGs. 17A-17G). IFN0-stimulated gene (ISG) expression is a hallmark of IFN activity7. To assess this transcriptional response. HEK-293 cells were treated for 24 hours with the optimal dose of the engineered IFN-beta or controls to saturate ISRE signaling. Total RNA was extracted and subjected to qPCR targeting the 02-micoglobulin and 2'-5' oligoadenylate synthetase 1 genes (OAS1). OAS1 expression was normalized to house-keeping gene, HPRT (FIGs. 18A-18F) Error bars represent mean + / - SD for n=3 biological replicate, ns = p = >0.05, * p = <0.05. Untreated cells were included as controls. These results demonstrate that some of the engineered proteins disclosed herein exhibited increased biological activity (“Biobetter”), than wildtype IFN-beta (SEQ ID NO: 1), while other engineered proteins exhibited similar biological activity (“Biosimilar”) to wildty pe IFN-beta (SEQ ID NO: 1). Notably, exemplary7 engineered proteins disclosed herein demonstrated reduced function or activity7. Representative engineered proteins for each category was tabulated in TABLE 10 below. TABLE 10. Properties of HIS-tagged engineered, non-naturally occurring proteins. Protein SEQ ID NO: Class 3xISRE Reporter Assay RT-PCR (OAS1 Biomarker) Flow-Cytometr y (3xISRE Reporter) Wildtype 1 WT Reference Reference Reference Wildtype 1 WT Reference Reference Reference K105N_E107T 2 WT Biobetter Biobetter Biobetter M117N 3 WT Biobetter Biobetter Biobetter E107N_E109T 4 WT Biosimilar Biosimilar Biosimilar S119N_H121T 5 GOF-like Biosimilar Biosimilar Biosimilar Y92NQ94T 137 Reduced Function Reduced Function Reduced Function Reduced Function RUN 6 WT Reduced Function NA NA N80Q_K99N_V101T 75 GOF-like NA NA NA Q72N 85 WT NA NA NA A89M 136 GOF-like Reduced Function NA NA L151N_N153T 119 WT NA NA NA RUN K105N E107T 343 Derivative Reduced Function NA NA R11N_M117N 344 Derivative Biosimilar Biosimilar Biosimilar Example 6 - Determining immunogenicity of non-naturally using Peripheral Blood Mononuclear Cells (PBMCs)

[00185] Cryopreserved PBMCs are harvested from healthy donors that have never been exposed to interferon therapy (FIG. 7). The non-naturally occurring proteins as described herein are taken up by antigen presenting cells (APCs) and presented to T cells in culture. If the non-naturally occurring protein is found to be immunogenic, which can lead to T cell activation and concurrent cytokine secretion. Immunogenicity assessment can then be performed by phenotyping the T cells using flow cytometry following stimulation with the non-naturally occurring proteins. Example 7 - Purification of untagged non-naturally occurring proteins

[00186] CHO cells are transfected with a mammalian expression plasmid to express HIS-tagged the non-naturally occurring proteins (FIG 8A-8B). A secretory signal is fused on the N-terminus of the 6xHIS tag to facilitate secretion of the protein into supernatant during cell culture. The secretory signal gets cleaved from the HIS-tagged non-naturally occurring protein during protein trafficking. Supernatant from transfected cells is loaded onto a Nickle-NTA (Ni-NTA) affinity column which binds the HIS residue of the HIS-tagged non-naturally occurring protein. Wash buffers are loaded into the column to wash away unbound material, and elution buffer is used to release the purified HIS-tagged non-naturally occurring protein from the column. Treatment with Enterokinase Light Chain proteinase is used to cleave the Enterokinase Light Chain Cleavage Site (EKCS) peptide sequence, separating the HIS-EKCS tag from the mature non-naturally occurring protein. The reaction is loaded into a purification column to remove the cleaved tag, resulting in purified untagged non-naturally occurring protein with no extra residues from the tags. Purified non-naturally occurring protein as described herein is checked for quality control using SDS-PAGE gels (to confirm purity and size), analytical size-exclusion (to assess the tag removal), Bradford assay (to measure concentration), and mass spectrometry (to confirm N-term. and C-term peptides) (FIG. 8A). Samples are lyophilized and resuspended in a pharmaceutically acceptable solvent. Amino acid sequence of 6xHIS and the EKCS protease site upstream of mature non-naturally occurring protein, with the cleavage position of enterokinase indicated by a dashed vertical line (FIG. 8B). Example 8 - Dose response of non-naturally occurring protein to compare EC50 values with an antiviral Cytopathic Effect (CPE) assay

[00187] A549 cells are plated in a 96-well plate and allowed to attach overnight. Once attached, cells are treated with serially-diluted purified non-naturally occurring proteins in parallel to a control wildtype IFN-beta, in triplicate (n=3) at varying concentrations covering the range of the dose curve. After 24 hours, media is replaced with media containing cytopathic encephalomyocarditis (EMC) virus at a concentration that causes 100% cell death in untreated cell cultures after 24 hours. ViralTox Gio (Promega) reagent is added 24 hours post-infection to measure ATP levels as an indicator of cytopathic effect, luminescence is quantified on a plate reader, and EC50 values for antiviral activity are determined for the non-naturally occurring proteins described herein (FIG. 9). Example 9 - Interferon-induced biomarker expression in human PBMCs

[00188] PBMCs (Peripheral Blood Mononuclear Cells), either fresh or cryopreserved, are treated with exemplary non-naturally occurring proteins as described herein in parallel to a control wildtype IFN-beta, in triplicate (n=3) (FIG. 10). After treatment, RNA is extracted and used for either RNAseq to sequence the transcriptome, or RT-PCR (for key biomarkers: B2M, 2',5'-OAS1, MxA) to assess the level of induction of interferon-stimulated genes between IFN leads compared to control wildtype IFN-beta. Example 10 - Prophylactic assessment of non-naturally occurring protein in Experimental Autoimmune Encephalomyelitis (EAE) mouse model for Multiple Sclerosis (MS)

[00189] Mice are immunized with MOG35-55-peptide at Day 0 to induce EAE. Also at Day 0, mice receive 30 mg / kg of purified, untagged IFN-pi lead candidate proteins in parallel to a control wildtype IFN-beta (10 mice per test subject: n=10) from Day 0-7 post-EAE induction (FIG. 11). Development of EAE symptoms are monitored through Day 36, covering the peak and chronic phases of EAE development. Splenocytes and spinal cord are harvested at Day 36, histopathology and FACS analysis of T cells and subsets of T cells is performed. Example 11 - Liver toxicity assessment of non-naturally occurring protein in mice

[00190] Mice are treated with purified, untagged IFN-[H lead candidate proteins in parallel to a control wildtype IFN-beta (3 mice per test subject; n=3) and monitored for up to 2 days posttreatment (FIG. 12). Blood samples are collected for liver function tests, and liver slides are prepared for pathology. Example 12 —Pharmacokinetic study of non-naturally occurring proteins

[00191] Healthy subjects are injected with a single subcutaneous dose of 60 mcg of non-naturally occurring protein (FIG. 13). Peak serum concentration (Cmax) with median time of peak serum concentration (Tmax), serum elimination half-life, and area under the serum concentration versus time curve from 0 to 96 hours are assessed. Example 13 -Immunogenicity study of non-naturally occurring proteins

[00192] The presence of neutralizing antibodies (NAb) to the non-naturally occurring protein is determined by collecting and analyzing serum pre-study and at 6-month time intervals during 2 years of the clinical trial (FIG. 14). Data reflects percentage of patients whose test results w ere considered positive for antibodies to the non-naturally occurring protein using antiviral cytopathic effect assay. Observed incidence of NAb positive in assay can be influence by several factors including sample handling, timing of sample collection, concomitant medications, and underlying disease.

[00193] While preferred embodiments of the present invention have been shown and described herein, it will be obvious to those skilled in the art that such embodiments are provided by way of example only. Numerous variations, changes, and substitutions will now occur to those skilled in the art without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention. It is intended that the following claims define the scope of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby.

Claims

1. An engineered interferon beta (IFN-beta) protein or a functional fragment thereof, comprising a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1, wherein the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208.

2. The engineered protein of claim 1, wherein the modification comprises an amino acid substitution, a deletion, or an insertion in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1.

3. The engineered protein of claim 2, wherein the modification is an amino acid substitution, and the amino acid substitution comprises at least one amino acid substitution in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1.

4. The engineered protein of claim 2, wherein the modification is a deletion of one or more amino acids in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1.

5. The engineered protein of claim 2, wherein the modification comprises an insertion of one or more amino acids in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1.

6. The engineered protein of claim 3, wherein the at least one amino acid substitution results in an increase in a biological activity of the engineered IFN-beta protein as compared to a biological activity of the wildtype protein.

7. The engineered protein of claim 3, wherein the at least one amino acid substitution results in a decrease in a biological activity of the engineered IFN-beta protein as compared to a biological activity of the wildtype protein.

8. The engineered protein of claim 7, wherein the increase in the biological activity of biological activity comprises an increase in signaling, an increase in protein expression of one or more biological targets, an increase in mRNA expression, or any combination thereof, as measured by an in vitro assay.

9. The engineered protein of claim 8, wherein the increase in signaling comprises an increase in IFNAR1-mediated signaling, as measured by an in vitro assay.

10. The engineered protein of claim 8, wherein the increase in signaling comprises an increase in IFNAR2-mediated signaling, as measured by an in vitro assay.

11. The engineered protein of claim 1, wherein the modification comprises:a. an amino acid substitution that results in a conversion of a motif in the amino acid sequence of SEQ ID NO: 1 that is not a glycosylation motif into a glycosylation motif, thereby introducing a glycosylation motif into the engineered protein;b. an amino acid substitution that results in a conversion of a glycosylation motif in the amino acid sequence of SEQ ID NO: 1 into a motif that is not a glycosylation motif, thereby eliminating a glycosylation motif from the engineered protein; orc. a combination of (a) and (b).

12. The engineered protein of claim 3, wherein the at least one amino acid substitution results in the conversion of 1, 2.

3. 4, or more motif(s) in the amino acid sequence of SEQ ID NO: 1 that are not glycosylation motifs into 1, 2, 3, 4, or more motif(s) that are glycosylation motif(s), thereby introducing 1, 2, 3, 4, or more glycosylation motif(s) into the engineered protein.

13. The engineered protein of claim 3, wherein the at least one amino acid substitution results in the conversion of 1, 2, 3, 4, or more glycosylation motif(s) in the amino acid sequence of SEQ ID NO: 1 into 1, 2, 3, 4, or more motif(s) that are not glycosylation motifs, thereby eliminating 1, 2, 3, 4, or more glycosylation motif(s) into the engineered protein.

14. The engineered protein of claim 11, wherein the glycosylation motif has a consensus sequence of N-X-S / T, wherein N is an Asparagine that is N-glycosylated at the amide bond in the side chain of the Asparagine, X is any amino acid except for proline, S is a serine, and T is a threonine.

15. The engineered protein of claim 1, wherein the modification comprises an amino acid substitution that results in a conversion of a motif in the amino acid sequence of SEQ ID NO: 1 that is not a site for a post-translational modification into a post-translational modification consensus sequence motif, thereby introducing a site for a post translational modification into the engineered protein.

16. The engineered protein of claim 15, wherein the post-translational modification consensus sequence motif is selected from the amino acid sequences in TABLE 5.

17. The engineered protein of claim 15, wherein the amino acid substitution results in the addition of a post-translational modification on the engineered protein at the site of the post-translational modification consensus sequence motif that is not present in the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1.

18. The engineered protein of claim 17, wherein the post-translational modification is selected from the group consisting of: glycosylation, phosphorylation, lipidation, glycation, proline isomerization, methylation, acetylation, hydroxylation sites, and proteolysis.

19. The engineered protein of any one of claims 1-18, wherein the engineered protein is glycosylated.

20. The engineered protein of any one of claims 1-18, wherein the engineered protein is aglycosylated.

21. The engineered protein of claim 1, wherein the engineered protein binds to an interferon-a / p receptor-1(IFN ARI) or an interferon-a / p receptor-2 (IFNAR2) heterodimer subunits at a level greater than or equal to the wildty pe IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by enzyme-linked immunosorbent assay (ELISA), or by measuring signaling activity of an Interferon Stimulated Response Element (ISRE)-Green fluorescent protein (GFP) reporter in a live human cell in vitro assay.

22. The engineered protein of claim 1, wherein the engineered protein binds to and activates an IFNAR1 or an IFNAR2 heterodimer subunits at a level greater than or equal to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by ELISA, or by measuring signaling activity of an ISRE-GFP reporter in a live human cell in vitro assay.

23. The engineered protein of claim 1, wherein the engineered protein binds to and inhibits an activity of an 1FNAR1 or an 1FNAR2 heterodimer subunits at a level greater than or equal to the wildtype IFN-beta protein as measured by ELISA, or by measuring signaling activity7 of an ISRE-GFP reporter in a live human cell in vitro assay.

24. The engineered protein of claim 1, wherein the engineered protein is an agonist of IFNAR1 or IFNAR2 subunits.

25. The engineered protein of claim 1, wherein the engineered protein binds and activates IFNAR1 or IFNAR2 at a similar level than the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by an ISRE-GFP reporter assay or an ISRE luciferase reporter assay.

26. The engineered protein of claim 1, wherein the engineered protein reduces an extracellular level of a pro-inflammatory cytokine, as compared to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by ELISA.

27. The engineered protein of claim 26, wherein the pro-inflammatory cytokine is a chemokine. an interferon (IFN), an interleukin (IL), a lymphokine, a tumor necrosis factor (TNF), or any combination thereof.

28. The engineered protein of claim 27, wherein the pro-inflammatory cytokine is IL-1Q, IL-6, IL-8, IL-9, IL-12, IL-15, IL-17, IL-18, IFN-y, TNF-a, TNF-0, or any combination thereof.

29. The engineered protein of claim 1, wherein the engineered protein increases an extracellular level of an anti-inflammatory cytokine, as compared to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by ELISA.

30. The engineered protein of claim 29, wherein the anti-inflammatory cytokine is a chemokine, an IFN, an IL, a lymphokine, a TNF, a Transforming growth factor (TGF), or any combination thereof.

31. The engineered protein of claim 30, wherein the anti-inflammatory cytokine is IL-4, IL-10, IL-11, IL-13, IFN-a, TGF-P, or any combination thereof.

32. The engineered protein of claim 1. wherein the engineered protein binds to IFNAR1 or IFNAR2 subunits at a level less than the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by ELISA.

33. The engineered protein of claim 1, wherein the engineered protein is an antagonist of IFNAR1 or IFNAR2 subunits.

34. The engineered protein of claim 33, wherein the engineered protein binds and inhibits an activity of the IFNAR1 or IFNAR2 at a level greater than wildty pe IFN-beta protein with the amino acid sequence of SEQ ID NO: 1, as measured by an ISRE GFP reporter assay or an ISRE luciferase reporter assay.

35. The engineered protein of any one of claims 1-34. wherein the engineered protein is a recombinant IFN-beta or a portion thereof.

36. The engineered protein of any one of claims 1-35, wherein the engineered protein is glycosylated, carboxylated. hydroxylated, sulfated, phosphorylated, albuminated, conjugated to a polyethylene glycol (PEG) moiety, or any combination thereof.

37. The engineered protein of claim 1, wherein a glycosylation site is introduced in the engineered protein that results in diminished binding of an antibody to the engineered protein, relative to an amount of binding of the antibody to the wildtype IFN-beta protein with the amino acid sequence of SEQ ID NO: 1.

38. The engineered protein of claim 1, wherein the modification results in a reduction of immunogenicity levels when administered to a subject, as compared to an immunogenicity7 level when the wildtype IFN-beta with the amino acid sequence of SEQ ID NO: 1 is administered to a subject.

39. The engineered protein of claim 38, wherein the reduction of immunogenicity levels comprises a reduced level of anti-drug antibody in the subject when the engineered protein is administered to the subject, as compared a level of anti-drug antibody when the wildtype IFN-beta with the amino acid sequence of SEQ ID NO: 1 is administered, as measured by ELISA on a sample obtained from the subject.

40. The engineered protein of claim 1, wherein the engineered protein, when administered to a subject in need thereof results in a reduction in a level of inflammation in a subject, relative to a level of inflammation in the subject prior to the administering.

41. The engineered protein of claim 40, wherein the reduction in the level of inflammation is determined by analyzing levels of an autoantibody, a C reactive protein (CRP), a proteolytic enzyme, an inflammatory mediator, a marker of ongoing inflammation, or any combination thereof, as measured by ELISA.

42. The engineered protein of claim 40, wherein the reduction in the level of inflammation in the subject results in a reduction in serum levels of C reactive protein as measured by high-sensitivity C-reactive protein (hs-CRP) test.

43. The engineered protein of claim 42, wherein the reduction in serum levels of C reactive protein comprises a plasma concentration in the subject of from about 0.1 mg / dL to about 2.9 mg / dL.

44. The engineered protein of claim 43, wherein the administering to the subject results in a reduction in blood levels of a pro-inflammatory cytokine as compared to the blood levels of one or more pro-inflammatory cytokines before the administering, as measured by ELISA.

45. The engineered protein of claim 1. wherein the functional fragment thereof comprises at least: 80%, 85%, 90%, 91%, 92%, 93%, 94% 95%, 96%, 97%, 98%, 99%, or 100% sequence identity' to any one of SEQ ID NO: 2 - SEQ ID NO: 208.

46. The engineered protein of claim 1, wherein the functional fragment thereof comprises at least: 80%, 85%, 90%, 91%, 92%, 93%, 94% 95%, 96%, 97%, 98%, 99%, or 100% sequence length to any one of SEQ ID NO: 2 - SEQ ID NO: 208.

47. The engineered protein of claim 1, wherein the functional fragment thereof comprises at least: 80%, 85%, 90%, 91%, 92%, 93%, 94% 95%, 96%, 97%, 98%, 99%, or 100% sequence homology’ to any one of SEQ ID NO: 2 - SEQ ID NO: 208.

48. The engineered protein of claim 1, wherein the engineered protein comprises an amino acid sequence that comprises a natural amino acid, or a non-natural amino acid.

49. A method of making a engineered IFN-beta protein in a cell, comprising introducing into the cell a vector encoding the engineered IFN-beta protein of any one of claims 1-4744, wherein the introducing results in expression of the engineered IFN-beta protein in the cell.

50. The method of claim 49, wherein the engineered IFN-beta protein is isolated from the cell.

51. The method of claim 49, wherein the vector comprises a mammalian expression plasmid.-102-52. The method of claim 49, wherein the engineered IFN-beta protein comprises a tag.

53. The method of claim 52, wherein the tag is CBP, FLAG, GST, Myc, poly-His, or V5.

54. The method of claim 52, wherein the tag comprises a cleavable tag or a non-cleavable tag.

55. A polynucleotide encoding an engineered interferon beta (IFN-beta) protein or a functional fragment thereof, comprising a modification in a wild type IFN-beta protein with an amino acid sequence of SEQ ID NO: 1, wherein the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208.

56. The polynucleotide of claim 55, wherein the polynucleotide comprises at least: 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of SEQ ID NO: 209 - SEQ ID NO: 342.

57. The polynucleotide of claim 55, wherein the polynucleotide comprises any one of SEQ ID NO: 209 - SEQ ID NO: 342.

58. The polynucleotide of claim 55, wherein the polynucleotide comprises at least: 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence length to any one of SEQ ID NO: 209 - SEQ ID NO: 342.

59. The polynucleotide of claim 55, wherein each codon in the polynucleotide can independently be replaced by a degenerate codon.

60. The polynucleotide of claim 55, wherein the polynucleotide comprise one or more degenerate codons.

61. The polynucleotide of claim 55, wherein the polynucleotide comprises a nucleic acid mimic.

62. The polynucleotide of claim 61, wherein the nucleic acid mimic comprises a phosphorothioate nucleic acid, a phosphoramidate nucleic acid, a morpholino nucleic acid, a hexitol nucleic acid (HNA), a peptide nucleic acid (PNA), or a locked nucleic acid (LNA).

63. The polynucleotide of claim 55, wherein one or more codons in the polynucleotide is modified.

64. The polynucleotide of claim 55, wherein one or more residues of an amino acid in the engineered IFN-beta protein is substituted for a non-natural amino acid.

65. A vector comprising the polynucleotide of any one of claims 55-64.

66. A pharmaceutical composition in unit dose form comprising:a. the engineered protein of any one of claims 1-48, or the polynucleotide of any one of claims 55-64; andb. a pharmaceutically acceptable excipient, carrier, or diluent.

67. The pharmaceutical composition of claim 66, wherein the pharmaceutical composition is encapsulated.

68. The pharmaceutical composition of claim 66, wherein the pharmaceutical composition is in the form of a liquid.

69. The pharmaceutical composition of any one of claims 66-68, wherein the pharmaceutical composition is formulated for local, systemic, or topical administration.

70. The pharmaceutical composition of any one of claims 66-69, wherein the pharmaceutical composition is formulated for oral, nasal, pulmonary , buccal, transdermal, subcutaneous, intraduodenal, enteral, parental, intravenous, or intramuscular administration.

71. The pharmaceutical composition of any one of claims 66-70, wherein the pharmaceutical composition is formulated for controlled-release.

72. The pharmaceutical composition of any one of claims 66-71, wherein the pharmaceutical composition is formulated for single-dosage administration.

73. A method of treating a disease in a subject, the method comprising: administering to the subject a therapeutically effective amount of a composition comprising (a) an engineered interferon beta (IFN-beta) protein or a functional fragment thereof; and (b) a pharmaceutically acceptable excipient, carrier, or diluent, wherein the engineered IFN-beta protein comprises a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1, and wherein the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208,.

74. The method of claim 73, wherein the disease is an infection.

75. The method of claim 74, wherein the infection is a viral infection.

76. The method of claim 75, wherein the viral infection is chronic Hepatitis B infection, or a chronic Hepatitis C infection.

77. The method of claim 73, wherein the disease is a demyelinating disorder.

78. The method of claim 77, wherein the demyelinating disorder is selected from the group consisting of Multiple Sclerosis (MS), Acute Disseminated Encephalomyelitis (ADEM), Acute Hemorrhagic Leucoencephalitis (AHLE), Balo’s disease (Concentric Sclerosis), Charcot-Marie-Tooth disease (CMT), Guillain-Barre Syndrome (GBS), HTLV-I Associated Myelopathy (HAM), and Neuromyelitis Optica (Devic’s Disease).

79. The method of claim 78, wherein the MS is relapsing remitting MS. non-relapsing MS. clinically isolated syndrome, and secondary progressive forms.

80. The method of any one of claims 73-79, wherein the subject is a human subject.

81. The method of any one of claims 73-80, wherein the administering is by parenchymal injection, intra-thecal injection, intra-ventricular injection, intra-cistemal injection,intratumoral injection, subcutaneous injection, intraperitoneal injection, a surgical route, or any combination thereof.

82. The method of claim 73, wherein the administering occurs from about once a day, about twice a day, about once a week, about twice a week, about once every two weeks, about once a month, about once every 3 months, about once every 6 months, about once every 9 months, to about once a year.

83. The method of claim 73, wherein the administering is at a dose selected from the group consisting of about 50 mg, about 75 mg, about 100 mg, about 150 mg, about 200 mg. about 300 mg, about 400 mg. about 500 mg. about 600 mg, about 700 mg, about 800 mg. about 900 mg, about 1000 mg, about 1200 mg, about 1500 mg, or about 2000 mg.

84. The method of claim 73, wherein the administering is at a dose selected from the group consisting of 1 mg / kg, 2 mg / kg, 3 mg / kg, 4 mg / kg, 5 mg / kg, 6 mg / kg, 7 mg / kg, 8 mg / kg, 9 mg / kg, and 10 mg / kg.

85. The method of claim 73, wherein the administering reduces a volume of a tumor in the subject.

86. The method of claim 73, wherein the administering inhibits or diminishes the growth a tumor in the subject.

87. The method of claim 73, wherein the method further comprises administering a second therapeutic.

88. The method of claim 87, wherein the second therapeutic is an antibody, a peptide, an antibody drug conjugate, a CAR-T, a cancer vaccine, or any combination thereof.

89. The method of claim 73, wherein the method further comprises administering an immunotherapy, and wherein the immunotherapy comprises a checkpoint inhibitor.

90. The method of claim 89, wherein the checkpoint inhibitor is a PD-1, a PD-L1, CTLA-4 inhibitor.

91. The method of claim 73, wherein the administering induces activation of an IFN-stimulated response element (ISRE) or a Gamma interferon activation site (GAS) element, wherein the activation of the ISRE or GAS induces an interferon-like activity as measured by a luciferase reporter assay.

92. A method of treating a disease or a condition in a subject, the method comprising: administering to the subject a therapeutically effective amount of engineered protein of any one of claims 1 -48, the polynucleotide of any one of claims 55-64, or the pharmaceutical composition of any one of claims 66-72, thereby treating the subject.

93. A method of treating a demyelinating disorder, the method comprising: administering to the subject a therapeutically effective amount of a composition comprising: (a) anengineered interferon beta (IFN-beta) protein or a functional fragment thereof, comprising a modification in a wildtype IFN-beta protein with an amino acid sequence of SEQ ID NO: 1, wherein the engineered IFN-beta protein comprises a polypeptide sequence of any one of SEQ ID NO: 2 - SEQ ID NO: 208; and (b) a pharmaceutically acceptable excipient, earner or diluent.

94. The method of claim 93, wherein the demyelinating disorder is selected from the group consisting of Multiple Sclerosis (MS), Acute Disseminated Encephalomyelitis (ADEM), Acute Hemorrhagic Leucoencephalitis (AHLE), Balo's disease (Concentric Sclerosis), Charcot-Marie-Tooth disease (CMT), Guillain-Barre Syndrome (GBS). HTLV-I Associated Myelopathy (HAM), and Neuromyelitis Optica (Devic’s Disease).

95. The method of claim 93, wherein the demyelinating disorder is a MS, wherein the MS is relapsing remitting MS, non-relapsing MS, clinically isolated syndrome, and secondary progressive forms.

96. A kit comprising the engineered protein of any one of claims 1-48, or the polynucleotide of any one of claims 55-64 or the pharmaceutical composition of any one of claims 6672, and a container.

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