Methods for predicting ancestral viral sequences and uses thereof

Abstract

Methods for predicting ancestral viral sequences and uses thereof are provided. Methods for predicting ancestral sequences of viruses or portions thereof are described. Predicted ancestral sequences of adeno-associated virus (AAV) capsid polypeptides are also described. The present disclosure also provides methods of gene transfer and methods of vaccinating a subject by administering a target antigen operably linked to an AAV capsid polypeptide.

Description

[0001] This application is a divisional application based on a patent application with an application date of October 10, 2014, a priority date of October 11, 2013, an application number of 201480065410.2, and an invention name of “Method for predicting ancestral virus sequences and use thereof”. Technical Field

[0002] Generally speaking, the present disclosure relates to viruses. Background Art

[0003] Evading and avoiding the neutralization or toxic immune response for gene therapy vectors is a major challenge faced by all gene transfer vector types. Up to now, gene transfer has been most effectively achieved using vectors (such as adenovirus and adeno-associated virus (AAV)) based on viruses circulating in humans and animals. However, if the subject has been naturally infected with the virus, then due to cellular and humoral immune responses, subsequent treatment with the vector based on the virus causes increased safety risks and reduced efficiency of gene transfer. Viral capsid antigens are primarily responsible for natural immunity and / or adaptive immunity against viral particles, but the polypeptides encoded by viral genes may also have immunogenicity. Summary of the Invention

[0004] The present disclosure describes methods for predicting and synthesizing ancestral viral sequences or portions thereof, and also describes viral particles containing such ancestral viral sequences. The methods described herein are applied to adeno-associated viruses (AAV); therefore, the present disclosure describes predicted ancestral AAV sequences and AAV viral particles containing such ancestral AAV sequences. The present disclosure also describes reduced seroprevalence exhibited by viral particles containing ancestral sequences relative to viral particles containing contemporary sequences.

[0005] In one aspect, the present disclosure includes an adeno-associated virus (AAV) capsid polypeptide, e.g., a synthetic and / or artificial AAV capsid polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO: 1, 3, 5, 7, 9, 11, 13, 15, and 17. In some embodiments, the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide exhibit a lower seroprevalence than the AAV2 capsid polypeptide or viral particles comprising the AAV2 capsid polypeptide, and the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide exhibits about the same or lower seroprevalence than the AAV8 capsid polypeptide or viral particles comprising the AAV8 capsid polypeptide. In some embodiments, the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide are neutralized to a lesser extent by human serum than the AAV2 capsid polypeptide or viral particles comprising the AAV2 capsid polypeptide, and the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide are neutralized to a similar or lower extent by human serum than the AAV8 capsid polypeptide or viral particles comprising the AAV8 capsid polypeptide. In some embodiments, the AAV capsid polypeptide is purified. The AAV capsid polypeptides provided herein can be encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, and 18.

[0006] In one aspect, the present disclosure provides nucleic acid molecules, e.g., synthetic and / or artificial nucleic acid molecules, encoding an adeno-associated virus (AAV) capsid polypeptide having a nucleic acid sequence selected from the group consisting of SEQ ID NOs: 2, 4, 6, 8, 10, 12, 14, 16, and 18. Also provided are vectors comprising such nucleic acids and host cells comprising such vectors.

[0007] In another aspect, the present disclosure provides purified viral particles comprising an AAV capsid polypeptide described herein. In some embodiments, the viral particle comprises a transgene.

[0008] In other aspects, the present disclosure provides adeno-associated virus (AAV) capsid polypeptides, e.g., synthetic and / or artificial AAV capsid polypeptides, having at least 95% (e.g., 97, 98, 99, or 100%) sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 19, 20, 21, 22, 23, 24, 25, and 26. In some embodiments, the AAV capsid polypeptides or viral particles comprising the AAV capsid polypeptides exhibit a lower seroprevalence than AAV2 capsid polypeptides or viral particles comprising the AAV2 capsid polypeptides, and the AAV capsid polypeptides or viral particles comprising the AAV capsid polypeptides exhibit about the same or lower seroprevalence than AAV8 capsid polypeptides or viral particles comprising the AAV8 capsid polypeptides. In some embodiments, the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide are neutralized by human serum to a lesser extent than AAV2 capsid polypeptide or viral particles comprising the AAV2 capsid polypeptide, and the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide are neutralized by human serum to a similar or lesser extent than AAV8 capsid polypeptide or viral particles comprising the AAV8 capsid polypeptide. In some embodiments, the AAV capsid polypeptide is purified.

[0009] In another aspect, the AAV capsid polypeptides described herein can be encoded by nucleic acid sequences as described herein. In one embodiment, the present disclosure provides nucleic acid molecules encoding adeno-associated virus (AAV) capsid polypeptides, wherein the nucleic acid molecules have at least 95% (e.g., 97, 98, 99, or 100%) sequence identity with the nucleic acids as set forth herein. The present disclosure also provides vectors comprising such nucleic acid molecules and host cells comprising such vectors.

[0010] In another aspect, the present disclosure provides a viral particle comprising at least one of the AAV capsid polypeptides described herein. In some embodiments, the viral particle comprises a transgene.

[0011] In certain aspects, the present disclosure provides methods of administering a viral particle as described herein to a subject in need of gene transfer or vaccination. In some embodiments, the viral particle exhibits a lower seropositivity rate than an AAV2 viral particle. In some embodiments, the viral particle exhibits a seropositivity rate that is approximately the same as or less than that of an AAV8 viral particle. In some embodiments, the viral particle is neutralized by human serum to a lesser extent than an AAV2 viral particle, and the AAV viral particle is neutralized by human serum to a similar or lesser extent than an AAV8 viral particle.

[0012] In one aspect, the present disclosure provides methods of administering a target antigen operably linked to an AAV capsid polypeptide as described herein to a subject in need of vaccination. In some embodiments, the AAV capsid polypeptide exhibits a lower seroprevalence than an AAV2 capsid polypeptide. In some embodiments, the AAV capsid polypeptide exhibits about the same or lower seroprevalence than an AAV8 capsid polypeptide. In some embodiments, the AAV capsid polypeptide is neutralized by human serum to a lesser extent than an AAV2 capsid polypeptide, and the AAV capsid polypeptide is neutralized by human serum to a similar or lesser extent than an AAV8 capsid polypeptide.

[0013] In another aspect, the present disclosure provides an in silico method for predicting the sequence of an ancestral virus or a portion thereof. Such methods generally include providing nucleotide or amino acid sequences from a plurality of contemporary viruses or portions thereof; aligning the sequences using a multiple sequence alignment (MSA) algorithm; modeling evolution to obtain a predicted ancestral phylogeny for a plurality of contemporary viruses or portions thereof; estimating the evolutionary probability of a particular nucleotide or amino acid residue at each position of the sequence at a phylogenic node of the predicted ancestral phylogeny and predicting the sequence of the ancestral virus or portion thereof based on the estimated probability at each position.

[0014] In some embodiments, one or more or all steps are carried out using a computer processor. In some embodiments, the MSA algorithm uses phylogenetic information to predict whether the gap in the comparison is the result of deletion or insertion. In some embodiments, the MSA algorithm is a probability comparison kit (Probabilistic Alignment Kit) (PRANK). In some embodiments, the model for evolutionary modeling is selected using Aikake information criterion (Aikake Information Criterion) (AIC). In some embodiments, the ancestral system of prediction is obtained using JTT model and gamma distribution model (Gamma distributionmodel) ("+G") and πi frequency calculation ("+F") generation. In some embodiments, the step of evolutionary modeling is carried out using JTT+G+F model. In some embodiments, the method includes synthesizing ancestral virus or its part based on the sequence predicted. In some embodiments, the method includes ancestral virus or its part being assembled into ancestral virus particle.

[0015] In some embodiments, the method further comprises screening the ancestral viral particles for at least one of: (a) replication; (b) gene transfer properties; (c) receptor binding; or (d) seropositivity. In some embodiments, the ancestral viral particles exhibit a lower seropositivity than viral particles assembled from at least one of a plurality of contemporary viruses or portions thereof. In some embodiments, viral particles assembled from the ancestral viral particles and at least one of a plurality of contemporary viruses or portions thereof are neutralized to a lesser extent by human serum. In some embodiments, the plurality of contemporary viruses or portions thereof belong to a family selected from the group consisting of adenovirus (AV), human immunodeficiency virus (HIV), lentivirus, lentivirus, herpes simplex virus (HSV), vaccinia virus, pox virus, influenza virus, respiratory syncytial virus, parainfluenza virus, and foamy virus.

[0016] Thus, the present disclosure provides ancestral viruses or portions thereof that exhibit reduced susceptibility to pre-existing immunity in contemporary human populations compared to contemporary viruses or portions thereof. Typically, the reduced susceptibility to pre-existing immunity exhibited by the ancestral virus or portion thereof in the current human population is reflected in reduced susceptibility to neutralizing antibodies.

[0017] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which the methods and material compositions belong. Although methods and materials similar or equivalent to the methods and materials described herein can be used in the practice or testing of the methods and material compositions, suitable methods and materials are described below. In addition, the materials, methods, and examples are illustrative only and are not intended to be limiting. All publications, patent applications, patents, and other references mentioned herein are incorporated herein by reference in their entirety.

[0018] The present invention includes the following embodiments:

[0019] 1. An adeno-associated virus (AAV) capsid polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, and 17.

[0020] 2. The AAV capsid polypeptide of embodiment 1, wherein the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide exhibit a lower seroprevalence than an AAV2 capsid polypeptide or viral particles comprising the AAV2 capsid polypeptide, and wherein the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide exhibit the same or lower seroprevalence than an AAV8 capsid polypeptide or viral particles comprising the AAV8 capsid polypeptide.

[0021] 3. The AAV capsid polypeptide of embodiment 1, wherein the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide are neutralized by human serum to a lesser extent than AAV2 capsid polypeptides or viral particles comprising AAV2 capsid polypeptides, and wherein the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide are neutralized by human serum to a similar or lesser extent than AAV8 capsid polypeptides or viral particles comprising AAV8 capsid polypeptides.

[0022] 4. The AAV capsid polypeptide of any one of embodiments 1-3, wherein the AAV capsid polypeptide is purified.

[0023] 5. The AAV capsid polypeptide of embodiment 1, which is encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16 and 18.

[0024] 6. A nucleic acid molecule encoding an adeno-associated virus (AAV) capsid polypeptide having a nucleic acid sequence selected from the group consisting of SEQ ID NO: 2, 4, 6, 8, 10, 12, 14, 16 and 18.

[0025] 7. A vector comprising the nucleic acid molecule of embodiment 6.

[0026] 8. A host cell comprising the vector of embodiment 7.

[0027] 9. A purified viral particle comprising the AAV capsid polypeptide of any one of embodiments 1-5.

[0028] 10. The purified viral particle of embodiment 9, further comprising a transgene.

[0029] 11. An adeno-associated virus (AAV) capsid polypeptide having at least 95% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NOs: 19, 20, 21, 22, 23, 24, 25, and 26.

[0030] 12. The AAV capsid polypeptide of embodiment 11, wherein the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide exhibit a lower seropositivity rate than an AAV2 capsid polypeptide or viral particles comprising an AAV2 capsid polypeptide, and wherein the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide exhibit the same or lower seropositivity rate than an AAV8 capsid polypeptide or viral particles comprising an AAV8 capsid polypeptide.

[0031] 13. The AAV capsid polypeptide of embodiment 11, wherein the AAV capsid polypeptide or viral particles comprising the AAV capsid polypeptide are neutralized by human serum to a lesser extent than AAV2 capsid polypeptides or viral particles comprising AAV2 capsid polypeptides, and wherein the AAV capsid polypeptides or viral particles comprising the AAV capsid polypeptide are neutralized by human serum to a similar or lesser extent than AAV8 capsid polypeptides or viral particles comprising AAV8 capsid polypeptides.

[0032] 14. The AAV capsid polypeptide of any one of embodiments 11-13, wherein the AAV capsid polypeptide is purified.

[0033] 15. The AAV capsid polypeptide of any one of embodiments 11-14, wherein the polypeptide has at least 99% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 19, 20, 21, 22, 23, 24, 25, and 26.

[0034] 16. The AAV capsid polypeptide of any one of embodiments 11-14, wherein the polypeptide has 100% sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 19, 20, 21, 22, 23, 24, 25, and 26.

[0035] 17. A viral particle comprising at least one of the AAV capsid polypeptides of any one of embodiments 11-16.

[0036] 18. The viral particle of embodiment 17, further comprising a transgene.

[0037] 19. A method for gene transfer or vaccination using a transgene, the method comprising

[0038] The viral particle of embodiment 10 or embodiment 18 is administered to a subject in need of gene transfer or vaccination, wherein the viral particle exhibits a lower seroprevalence than AAV2 viral particles.

[0039] 20. The method of embodiment 19, wherein the viral particles exhibit approximately the same or less seroprevalence as AAV8 viral particles.

[0040] 21. The method of embodiment 19, wherein the viral particles are neutralized by human serum to a lesser extent than AAV2 viral particles, and wherein the AAV viral particles are neutralized by human serum to a similar or lesser extent than AAV8 viral particles.

[0041] 22. A method for vaccinating a subject, the method comprising

[0042] A target antigen operably linked to an AAV capsid polypeptide of embodiment 1 or embodiment 11 is administered to a subject in need of vaccination, wherein the AAV capsid polypeptide exhibits a lower seroprevalence than an AAV2 capsid polypeptide.

[0043] 23. The method of embodiment 22, wherein the AAV capsid polypeptide exhibits approximately the same or less seroprevalence as the AAV8 capsid polypeptide.

[0044] 24. The method of embodiment 22, wherein the AAV capsid polypeptide is neutralized by human serum to a lesser extent than AAV2 capsid polypeptide, and the AAV capsid polypeptide is neutralized by human serum to a similar or lesser extent than AAV8 capsid polypeptide.

[0045] 25. An in silico method for predicting the sequence of an ancestral virus or a portion thereof, the method comprising:

[0046] Providing nucleotide or amino acid sequences from multiple contemporary viruses or parts thereof;

[0047] The sequences were aligned using the Multiple Sequence Alignment (MSA) algorithm;

[0048] modeling evolution to obtain a predicted ancestral phylogeny of the plurality of contemporary viruses or portions thereof;

[0049] estimating the evolutionary probability of a particular nucleotide or amino acid residue at each position of the sequence at a phylogenetic node of the predicted ancestral phylogeny, and

[0050] The sequence of the ancestral virus or a portion thereof is predicted based on the estimated probability at each position.

[0051] 26. The method of embodiment 25, wherein all steps are performed using a computer processor.

[0052] 27. The method of embodiment 25 or embodiment 26, wherein the MSA algorithm uses phylogenetic information to predict whether a gap in the alignment is the result of a deletion or an insertion.

[0053] 28. The method of embodiment 27, wherein the MSA algorithm is Probabilistic Alignment Kit (PRANK).

[0054] 29. The method of any one of embodiments 25 to 28, wherein the model for modeling evolution is selected using the Aikake Information Criterion (AIC).

[0055] 30. The method of any one of embodiments 25 to 28, wherein the predicted ancestral phylogeny is obtained using a JTT model and a gamma distribution model ("+G") and a frequency calculation of πi ("+F").

[0056] 31. The method of embodiment 25, wherein the step of modeling evolution is performed using a JTT+G+F model.

[0057] 32. The method of any one of embodiments 25 to 31, further comprising synthesizing the ancestral virus or a portion thereof based on the predicted sequence.

[0058] 33. The method of embodiment 32, further comprising assembling the ancestral virus or a portion thereof into an ancestral virus particle.

[0059] 34. The method of embodiment 33, further comprising screening the ancestral viral particles for at least one of: (1) replication; (b) gene transfer properties; (c) receptor binding; or (d) seropositivity.

[0060] 35. The method of embodiment 34, wherein the ancestral viral particles exhibit a lower seroprevalence than viral particles assembled from at least one of the plurality of contemporary viruses or portions thereof.

[0061] 36. The method of embodiment 34, wherein the ancestral viral particle is neutralized by human serum to a lesser extent than viral particles assembled from at least one of the plurality of contemporary viruses or portions thereof.

[0062] 37. The method of embodiment 25, wherein the plurality of contemporary viruses or portions thereof belong to a family selected from the group consisting of adenovirus (AV), human immunodeficiency virus (HIV), lentivirus, lentivirus, herpes simplex virus (HSV), vaccinia virus, pox virus, influenza virus, respiratory syncytial virus, parainfluenza virus, and foamy virus. BRIEF DESCRIPTION OF THE DRAWINGS

[0063] This patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0064] Figure 1 is a schematic diagram showing the relationship between ancestral virus / contemporary virus infection and ancestral host / contemporary host immune response.

[0065] Figure 2 Figures a to d are a series of schematic diagrams showing an example of the ancestral reconstruction process. The data shown are excerpts from the complete dataset and represent residues 564-584 (AAV2-VP1 numbering).

[0066] Figure 3 Shown together are phylogenetic trees of contemporary AAV sequences generated using the methods described herein.

[0067] Figure 4 An alignment of ancestral AAV VP1 polypeptides is shown together.

[0068] Figure 5A and 5B An alignment of functional ancestral AAV VP1 polypeptides and contemporary AAV VP1 polypeptides are shown together.

[0069] Figure 6 is a gel electrophoresis image demonstrating that the ancestral AAV VP1 sequence is transcribed and alternatively spliced ​​in a manner similar to contemporary AAV VP1 sequences.

[0070] Figure 7 is a graph showing luciferase activity in HEK293 cells transduced with the ancestral AAV vector.

[0071] Figure 8 is a graph showing that ancestral AAV vectors are more resistant to IVIG neutralization than contemporary AAV vectors.

[0072] Figure 9 Graph showing sequence comparison between the Anc80 library and Anc80L65 (% from the diagonal upwards, # of amino acid differences below).

[0073] Figure 10 Figures AD are images of experimental results demonstrating that Anc80L65 can assemble and produce high-titer particles. Panel A shows that Anc80L65 can produce vector yields comparable to AAV2; Panel B is a TEM image of viral particles containing Anc80L65; Panel C shows viral particles containing Anc80L65 that can produce AAV cap VP1, 2, and 3 proteins based on SDS-PAGE gel under denaturing conditions; and Panel D shows a Western blot of Anc80L65 using AAV capsid antibody B1.

[0074] Figure 11 AC are images of experimental results demonstrating that Anc80L65 is able to infect HEK293 cells in vitro relative to AAV2 and / or AAV8 controls using GFP as a readout (Panel A) or luciferase (Panel B), and is effectively targeted to the liver after IV injection of AAV encoding the nuclear LacZ transgene (top row, Panel C: liver), after direct IM injection of AAV encoding GFP (middle row, Panel C: muscle), and after subretinal injection of AAV encoding GFP (bottom row, Panel C: retina).

[0075] Figure 12 Figures AD show that Anc80L64 is minimally seropositive in human populations using IVIg (Panel A) or sera from the Belgian population (Panel B), sera from the Boston population (Panel C), or sera from Cynomolgus macaques (Panel D).

[0076] Figure 13A and 13B is a sequence identity matrix generated using MAFFT, which shows the amino acid sequence of the VP1 protein of the ancestral vector aligned with the amino acid sequence of the VP1 protein of a representative extant AAV ( Figure 13A ), and the amino acid sequence of the VP3 protein of the ancestral vector aligned with the amino acid sequence of the VP3 protein of representative existing AAVs ( Figure 13B ).

[0077] Figure 14Graph showing the production of AAV vectors in triplicate at a small scale (6-well dish). Crude virus was assessed by qPCR to determine the absolute production of each vector.

[0078] Figure 15 is a table showing the titer of each vector averaged and compared to the titer of AAV8.

[0079] Figure 16 : is a photograph showing the results of an experiment in which 1.9E3 GC / cell of each vector was added to HEK293 cells (except Anc126, in which an MOI of 2.5E2-3.1E2 GC / cell was achieved). After 60 hours, infectivity was assessed by fluorescence microscopy.

[0080] Figure 17 is a graph showing the results of an experiment where the cleavage from Figure 16 The same cells were cultured and luciferase expression was determined. Figure 16 As in the previous study, the titer of Anc126 was not controlled with other vectors, but the MOI ranged from 2.5E2 to 3.1E2 GC / cell.

[0081] Figure 18 The table shows the luminescence of cells transduced with each vector, which was averaged and compared to the luminescence of AAV8.

[0082] Figure 19 The figure provides a summary of in vitro experiments determining the relative production and infectivity of the ancestral AAV vectors described herein. DETAILED DESCRIPTION

[0083] Gene transfer (for experimental or therapeutic purposes) relies on a vector or vector system to shuttle genetic information into target cells. The vector or vector system is considered to be a major determinant of the efficiency, specificity, host response, pharmacology and longevity of the gene transfer reaction. Currently, the most efficient and effective way to achieve gene transfer is through the use of vectors or vector systems based on viruses that have been rendered replication-defective.

[0084] However, seroprevalence studies indicate that a significant proportion of the global human population has been pre-exposed (e.g., by natural infection) to a large number of viruses currently used in gene transfer, and therefore has pre-existing immunity. It is known that neutralizing antibodies against viral vectors in these pre-exposed individuals sometimes significantly limit the extent of gene transfer, or even redirect the virus away from the target. See, for example, Calcedo et al., (2009, J. Infect. Dis., 199: 381-90) and Boutin et al., (2010, Human Gene Ther., 21: 704-12). Therefore, the present disclosure is based on the understanding that ancestral viruses or parts thereof exhibit reduced susceptibility to pre-existing immunity (e.g., reduced susceptibility to neutralizing antibodies) in current human populations compared to contemporary viruses or parts thereof.

[0085] Figure 1 is a schematic diagram showing the relationship between ancestral and contemporary viral infections and ancestral and contemporary host immune responses. Figure 1 The present invention shows how ancestral AAV can be refractory to contemporary pre-existing immunity. It is speculated that contemporary existing viruses (Vc) evolved from ancestral species (Vanc) mainly through immune escape mechanisms under the evolutionary pressure of host immunity. Each of these species Vanc and Vc has the ability to induce adaptive immunity, including B cell and T cell immunity (Ianc and Ic, respectively). It is hypothesized and confirmed herein that the immunity induced by contemporary viruses does not necessarily cross-react with ancestral virus species, which may be substantially different from existing viruses in terms of epitope composition.

[0086] The present disclosure provides a method for predicting the sequence of an ancestral virus or a portion thereof. One or more ancestral virus sequences predicted using the methods described herein can be generated and assembled into viral particles. As demonstrated herein, viral particles assembled from predicted ancestral virus sequences can exhibit smaller, sometimes significantly smaller, seropositive rates than current contemporary viral particles. Therefore, the ancestral virus sequences disclosed herein are suitable for use in vectors or vector systems for gene transfer.

[0087] Methods for predicting and synthesizing ancestral viral sequences

[0088] In order to predict the ancestral viral sequence, first nucleotide or amino acid sequences are compiled from multiple contemporary viruses or parts thereof. Although the method described herein is exemplified by adeno-associated virus (AAV) capsid sequences, the same method can be applied to other sequences (e.g., whole genome, rep sequences, ITR sequences) from AAV or any other virus or part thereof. Viruses other than AAV include, but are not limited to, adenovirus (AV), human immunodeficiency virus (HIV), retroviruses, slow viruses, herpes simplex virus (HSV), measles virus, vaccinia virus, poxvirus, influenza virus, respiratory syncytial virus, parainfluenza virus, foamy virus, or any other virus where pre-existing immunity is problematic.

[0089] Sequences from as few as two contemporary viruses or portions thereof can be used, however, it will be appreciated that a greater number of sequences of contemporary viruses or portions thereof are desirable in order to encompass as much of the landscape of contemporary sequence diversity as possible, and also because a greater number of sequences can increase the predictive power of the algorithms described and used. For example, sequences from 10 or more contemporary viruses or portions thereof can be used, sequences from 50 or more contemporary viruses or portions thereof can be used, and sequences from 100 or more contemporary viruses or portions thereof can be used.

[0090] Such sequences can be obtained, for example, from many public databases, including but not limited to GenBank, UniProt, EMBL, International Nucleotide Sequence Database Collaboration (INSDC) or European Nucleotide Archive. Additionally or alternatively, such sequences can be obtained from databases specific to particular organisms (e.g., HIV databases). Contemporary sequences can correspond to the entire genome, or only a portion of the genome can be used, such as but not limited to sequences encoding one or more components of viral capsids, replication proteins, or ITR sequences.

[0091] Next, contemporary sequences were aligned using the Multiple Sequence Alignment (MSA) algorithm. Figure 2(a) is a schematic diagram showing the alignment of multiple sequences. MSA algorithms are well known in the art and are generally designed to be applicable to data sets of different sizes and different inputs (e.g., nucleic acids or proteins), as well as to align sequences in a particular manner (e.g., dynamic programming, progressive, heuristic) and to apply different scoring schemes in the alignment (e.g., matrix-based or consistency-based, e.g., minimum entropy, pairwise sum, similarity matrix, gap score). Well-known MSA algorithms include, for example, ClustalW (Thompson et al., 1994, Nuc. Acids Res., 22:4673-90), Kalign (Lassman et al., 2006, Nuc. Acids Res., 34:W596-99), MAFFT (Katoh et al., 2005, Nuc. Acids Res., 33:511-8), MUSCLE (Edgar, 2004, BMC Bioinform., 5:113), and T-Coffee (Notredame et al., 2000, J. Mol. Biol., 302:205-17).

[0092] As described herein, one of the main features when selecting the MSA algorithm for the method described herein is the mode of the gap in the algorithm processing comparison. Penalty values ​​(penalty value) can be assigned to the gap in the sequence alignment, which depends on or does not depend on the gap size. In the present method, preferably, with the biased, non-systematic processing of the gap caused by, for example, insertion and / or deletion, in contrast, the MSA algorithm used in the method described herein applies phylogenetic information to the gap in the comparison to predict whether it is the result of deletion or insertion. The suitable method for processing gaps in comparison and evolutionary analysis is described in Loytynoja and Goldman, 2008, Science, 320:1632-5, and the commercially available algorithm applied to the comparison in a manner suitable for the method described herein is Probabilistic Alignment Kit (PRANK; Goldman Group Software; Loytynoja and Goldman, 2005, PNAS USA, 102:10557-62) and a modification of the PRANK algorithm.

[0093] Evolutionary models are then applied to the resulting alignments to obtain predicted ancestral phylogenies (see Figure 2(b)). There are many evolutionary models available in the art, each of which applies a slightly different matrix of substitution rates to amino acids. Algorithms that apply evolutionary models include, but are not limited to, the Dayhoff model (e.g., PAM120, PAM160, PAM250; Dayhoff et al., 1978, In Atlas of Protein Sequence and Structure (ed. Dayhoff), pp. 345-52, National Biomedical Research Foundation, Washington DC), the JTT model (Jones et al., 1992, Comp. Appl. Biosci., 8: 275-82), the WAG model (Whelan and Goldman, 2001, Mol. Biol. Evol., 18: 691-9), and the Blosum model (e.g., Blosum45, Blosum62, Blosum80; Henikoff and Henikoff, 1992, PNAS USA, 89: 10915-9).

[0094] In addition, the constraints imposed on evolutionary models by structure and function can themselves be modeled, for example, by considering that some positions are invariant ("+I"; Reeves, 1992, J. Mol. Evol., 35:17-31), some positions undergo change at different rates ("+G"; Yang, 1993, Mol. Biol. Evol., 10:1396-1401), and / or the equilibrium frequencies of nucleotides or amino acids are the same as those in the alignment ("+F"; Cao et al., 1994, J. Mol. Evol., 39:519-27).

[0095] The fitness of one or more evolutionary models can be assessed using the Aikake Information Criterion (AIC; Akaike, 1973, In Second International Symposium on Information Theory, Petrov and Csaki, eds., pp 267-81, Budapest, Akademiai Kiado), the Bayesian Information Criterion (BIC; Schwarz, 1978, Ann. Statist. 6:461-4), or variations or combinations thereof. In addition, AIC, BIC, or variations or combinations thereof can be used to assess the relative importance of including one or more parameters (e.g., the constraints discussed above) in the evolutionary model.

[0096] As explained in the Examples section below, based on the lowest AIC, ProTest3 (Darriba et al., 2011, Bioinformatics, 27(8): 1164-5) can be used to determine that the JTT+G+F algorithm is the most suitable model for AAV evolution. Those skilled in the art will understand that the JTT+G+F algorithm can also be used to predict ancestral viral sequences that differ from AAV capsid polypeptides. However, those skilled in the art will also understand that depending on the data set and fitness score, a different evolutionary model may be more appropriate.

[0097] Once an evolutionary model has been selected and its fitness has been determined, a phylogenetic tree of a viral sequence or its part can be constructed. Construction of a phylogenetic tree is known in the art, and maximum likelihood method (maximum likelihood method) is generally used, such as those performed by PhyML (Guindon and Gascuel, 2003, Systematic Biology, 52:696-704)), MOLPHY (Adachi and Hasegawa, 1996, ed.Tokyo Institute of Statistical Mathematics), BioNJ (Gascuel, 1997, Mol.Biol.Evol., 14:685-95) or PHYLIP (Felsenstein, 1973, Systematic Biology, 22:240-9). It will be appreciated by those skilled in the art that the balance between computational complexity and goodness of fit (goodness of fit) in the model of amino acid substitution is desirable.

[0098] As needed, the significance of the phylogenetic tree can be assessed. Many statistical methods are available and routinely used to assess the significance of models, including but not limited to bootstrap, jackknife, cross-validation, permutation test, or combinations or variations thereof. Significance can also be assessed using, for example, an approximate likelihood-ratio test (aLRT; Anisimova and Gascuel, 2006, Systematic Biology, 55:539-52).

[0099] At any phylogenetic node in a phylogeny (e.g., an internal phylogenetic node), the sequence can be reconstructed by estimating the probability of evolution of a particular nucleotide or amino acid residue at each position in the sequence (e.g., Figure 2 (c)). A phylogenetic node refers to an intermediate evolutionary branch point within a predicted ancestral phylogeny. As used herein, "probability of evolution" refers to the probability of a particular nucleotide or amino acid existing at a particular position based on an evolutionary model, in contrast to a model that does not consider, for example, evolutionary shift in codon selection. Any number of maximum likelihood methods can be used, including but not limited to, phylogenetic analysis by maximum likelihood (Phylogenetic Analysis by Maximum Likelihood) (PAML; Yang, 1997, Comp. Applic. BioSci., 13: 555-6) or phylogenetic analysis using parsimony (PAUP; Sinauer Assoc., Inc., Sunderland, MA) that takes into account exemplary models of the evolutionary probability of a particular nucleotide or amino acid residue at a particular position.

[0100] Based on the estimated evolutionary probabilities of specific nucleotide or amino acid residues at each position, the predicted sequences of the ancestral virus or portions thereof can be assembled to form complete or partial synthetic nucleic acid or polypeptide sequences. As desired, the probability of any residue along the nodes being in a given state at a given node can be calculated, and any position along the sequence having a calculated posterior probability below a specific threshold can be identified ( Figure 2 In this way, an ancestral scaffold sequence can be generated that includes variations at those positions with a probability below a certain threshold.

[0101] If the ancestral sequence predicted using the methods herein is a nucleic acid sequence, then the sequence can be codon optimized so that it can be efficiently translated into an amino acid sequence. Codon selection tables for different organisms are known in the art. Optionally, however, the codon selection table can be designed based on one or more contemporary sequences having identity (e.g., at least 90% sequence identity) with the ancestral scaffold sequence, and the ancestral sequence as described herein can be codon optimized for the codon selection of mammals (e.g., humans).

[0102] Any or all of the steps outlined herein for predicting ancestral viral sequences can be performed or simulated on a computer (e.g., in silico) using a processor or microprocessor.

[0103] Ancestral adeno-associated virus (AAV) scaffold sequence

[0104] The methods described herein were applied to adeno-associated virus (AAV) using contemporary capsid sequences (described in detail in the Examples below). AAV is widely recognized as a therapeutic gene transfer vector and genetic vaccine vector, but exhibits high seroprevalence in the human population. Using the methods described herein, contemporary AAV sequences (see Figure 3 ) assembles a phylogenetic tree and obtains predicted ancestral scaffold sequences at designated phylogenetic nodes (Table 1). As used herein, an ancestral scaffold sequence refers to a sequence constructed using the methods described herein (e.g., using evolutionary probability and evolutionary modeling) and known to not exist in nature. As used herein, an ancestral scaffold sequence is different from a consensus sequence, which is typically constructed using the frequency of nucleotides or amino acid residues at specific positions.

[0105] Table 1

[0106]

[0107] The scaffold sequence of the Anc80 polypeptide is shown in SEQ ID NO: 1, which is encoded by the scaffold sequence of the Anc80 nucleic acid shown in SEQ ID NO: 2. The scaffold sequence of Anc80 contains 11 positions at which either of two residues is possible. Thus, the Anc80 scaffold sequence represents 2048 (2 11 ) different sequences.

[0108] To demonstrate the effectiveness of the methods described herein for predicting ancestral sequences of viruses or portions thereof, a library of 2048 predicted ancestral sequences at the AAV Anc80 node was generated and, as described herein, demonstrated to form viable viral particles that exhibited lower seroprevalence, in some cases significantly lower seroprevalence, than viral particles assembled with contemporary capsid polypeptides.

[0109] Methods for preparing ancestral virus particles

[0110] After having obtained the ancestral sequence of the prediction of virus or its part, can produce (for example synthesize) actual nucleic acid molecule and / or polypeptide.The method for producing artificial nucleic acid molecule or polypeptide based on the sequence that for example computer obtains is known in the art, and comprises for example chemical synthesis or recombinant cloning.Other methods for producing nucleic acid molecule or polypeptide are known in the art, and are discussed in more detail below.

[0111] In case ancestral polypeptide has been produced, or in case ancestral nucleic acid molecule has been produced and expressed to produce ancestral polypeptide, ancestral polypeptide can be assembled into ancestral virus particle using, for example, packaging host cell. Components of virus particle (for example, rep sequence, cap sequence, terminal inverted repeat (ITR) sequence) can be transiently or stably introduced into packaging host cell using one or more vectors as described herein. One or more components of virus particle can be based on the ancestral sequence of prediction as described herein, and remaining components can be based on contemporary sequence. In some cases, whole virus particle can be based on the ancestral sequence of prediction.

[0112] Such ancestral viral particles can be purified using conventional methods. As used herein, "purified" viral particles refer to viral particles that have been removed from the components of the mixture from which they were prepared, such as, but not limited to, viral components (e.g., rep sequences, cap sequences), packaging host cells, and partially or incompletely assembled viral particles.

[0113] Once assembled, the ancestral virus particles can be screened for, for example, the ability to replicate; Gene transfer properties; Receptor binding capacity; and / or seropositivity in a population (e.g., a human population). Determining whether a virus particle can replicate is conventional in the art and typically includes infecting a host cell with a certain amount of virus particles and determining whether the number of virus particles increases over time. Determining whether a virus particle is able to perform gene transfer is also conventional in the art and typically includes infecting a host cell with a virus particle containing a transgene (e.g., a detectable transgene, such as a reporter gene, discussed in more detail below). After viral infection and removal, the presence or absence of the transgene in the host cell can be assessed. Determining whether a virus particle is bound to its receptor is conventional in the art, and such methods can be performed in vitro or in vivo.

[0114] Determining the seroprevalence of viral particles is routinely performed in the art and generally includes using immunoassays to determine the positive rate (prevalence) of one or more antibodies in a sample (e.g., a blood sample) from a specific population of an individual. Seroprevalence is understood in the art to refer to the ratio of subjects who are seropositive (i.e., exposed to a specific pathogen or immunogen) in a population, and is calculated as the number of subjects who produce antibodies against a specific pathogen or immunogen in a population divided by the total number of individuals in the population being examined. Immunoassays are well known in the art and include, but are not limited to, immune dot traces, Western blots, enzyme immunoassays (EIA), enzyme-linked immunosorbent assays (ELISA), or radioimmunoassays (RIA). As noted herein, ancestral viral particles exhibit smaller seroprevalence than contemporary viral particles (i.e., viral particles assembled using contemporary viral sequences or their parts). By way of example only, see Xue et al. (2007, Am. J. Obstet. Gynecol., 196:43.e1-6); Paul et al. (1994, J. Infect. Dis., 169:801-6); Sauerbrei et al. (2011, Eurosurv., 16(44):3); and Sakhria et al. (2013, PLoS Negl. Trop. Dis., 7:e2429), each of which determines the seroprevalence of a particular antibody in a given population.

[0115] As described herein, ancestral virus particles are neutralized by the immune system of an individual (e.g., a patient) to a lesser extent than contemporary virus particles. Several methods for determining the extent of neutralizing antibodies in serum samples are available. For example, neutralizing antibody assays measure the titer of antibody concentrations in experimental samples that neutralize infection by 50% or more compared to control samples without antibodies. See also Fisher et al. (1997, Nature Med., 3:306-12) and Manning et al. (1998, Human Gene Ther., 9:477-85).

[0116] For the ancestral AAV capsid polypeptides exemplified herein, the seroprevalence and / or degree of neutralization can be compared, for example, to AAV8 capsid polypeptides or viral particles comprising AAV8 capsid polypeptides, or AAV2 capsid polypeptides or viral particles comprising AAV2 capsid polypeptides. It is generally understood in the art that AAV8 capsid polypeptides or viral particles exhibit what is considered a lower seroprevalence and resulting neutralization in human populations, while AAV2 capsid polypeptides or viral particles exhibit what is considered a higher seroprevalence and resulting neutralization in human populations. Obviously, the specific seroprevalence will depend on the population examined and the immunological method used, but it has also been reported that AAV8 exhibits a seroprevalence of about 22% to about 38%, while AAV2 exhibits a seroprevalence of about 43.5% to about 72%. See, e.g., Boutin et al., 2010, "Prevalence of serum IgG and neutralizing factors against AAV types 1,2,5,6,8and 9 in the healthy population: implications for gene therapy using AAV vectors," Hum. GeneTher., 21:704-12. See also, Calcedo et al., 2009, J. Infect. Dis., 199:381-90.

[0117] Predicted ancestral nucleic acid and polypeptide sequences of adeno-associated virus (AAV)

[0118] A variety of different clones from the library encoding ancestral capsid polypeptides predicted from the Anc80 node were sequenced, and the amino acid sequences of representative AAV predicted ancestral capsid polypeptides are shown in SEQ ID NO: 19 (Anc80L27); SEQ ID NO: 20 (Anc80L59); SEQ ID NO: 21 (Anc80L60); SEQ ID NO: 22 (Anc80L62); SEQ ID NO: 23 (Anc80L65); SEQ ID NO: 24 (Anc80L33); SEQ ID NO: 25 (Anc80L36); and SEQ ID NO: 26 (Anc80L44). Those skilled in the art will appreciate that the nucleic acid sequence encoding each amino acid sequence can be readily determined.

[0119] In addition to the predicted ancestral capsid polypeptides having the sequence set forth in SEQ ID NO: 19, 20, 21, 22, 23, 24, 25, or 26, polypeptides are provided that have at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to the predicted ancestral capsid polypeptides having the sequence set forth in SEQ ID NO: 19, 20, 21, 22, 23, 24, 25, or 26. Similarly, nucleic acids are provided that have at least 95% sequence identity (e.g., at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity) to nucleic acids encoding ancestral capsid polypeptides (i.e., have at least 95% sequence identity).

[0120] When calculating percent sequence identity, two sequences are compared and the number of identical matches of the nucleotides or amino acid residues between the two sequences is determined. The number of identical matches is divided by the length of the comparison area (that is, the number of nucleotides or amino acid residues compared), and multiplied by 100 to obtain a percent sequence identity value. It should be understood that the length of the comparison area can be a portion of one or two sequences up to the full-length size of the shortest sequence. It should also be understood that a single sequence can be compared with more than one other sequence and, therefore, can have different percent sequence identity values ​​relative to each comparison area.

[0121] The comparison of two or more sequences can be performed to determine the percent sequence identity using the algorithm described by Altschul et al. (1997, Nucleic Acids Res., 25:3389-3402), which is incorporated into the BLAST (Basic Local Alignment Search Tool) program, available on the World Wide Web at ncbi.nlm.nih.gov. A BLAST search can be performed to determine the percent sequence identity between a sequence (nucleic acid or amino acid) and any other sequence or portion thereof compared using the algorithm of Altschul et al. BLASTN is a program for comparing and comparing the identity between nucleic acid sequences, while BLASTP is a program for comparing and comparing the identity between amino acid sequences. When using the BLAST program to calculate the percent identity between a sequence and another sequence, the default parameters of each program are generally used.

[0122] Representative alignments are shown in Figure 4 and Figure 5A and 5B middle. Figure 4An alignment of the ancestral AAV VP1 capsid polypeptides designated Anc80L65 (SEQ ID NO: 23), Anc80L27 (SEQ ID NO: 19), Anc80L33 (SEQ ID NO: 24), Anc80L36 (SEQ ID NO: 25), Anc80L44 (SEQ ID NO: 26), Anc80L59 (SEQ ID NO: 20), Anc80L60 (SEQ ID NO: 21), and Anc80L62 (SEQ ID NO: 22) is shown. Figure 4 The alignment shown in confirms predicted variations at each of the 11 sites, as well as a single non-synonymous mutation at position 609E of Anc80L60 (SEQ ID NO: 21), which may be a clonal artifact. Figure 5A and 5B An alignment between ancestral AAV VP1 capsid polypeptides (Anc80L65 (SEQ ID NO:23), Anc80L27 (SEQ ID NO:19), Anc80L33 (SEQ ID NO:24), Anc80L36 (SEQ ID NO:25), Anc80L60 (SEQ ID NO:21), Anc80L62 (SEQ ID NO:22), Anc80L44 (SEQ ID NO:26), and Anc80L59 (SEQ ID NO:20)) and contemporary AAV VP1 capsid polypeptides (AAV8 (SEQ ID NO:27), AAV9 (SEQ ID NO:28), AAV6 (SEQ ID NO:29), AAV1 (SEQ ID NO:30), AAV2 (SEQ ID NO:31), AAV3 (SEQ ID NO:32), AAV3B (SEQ ID NO:33), and AAV7 (SEQ ID NO:34) is shown. Figure 5A and 5B Alignment of the ancestral AAV sequences showed that the ancestral AAV sequences had between approximately 85% and 91% sequence identity with contemporary AAV sequences.

[0123] Also provided are vectors containing nucleic acid molecules encoding polypeptides. Vectors (including expression vectors) are commercialized or can be produced by recombinant technology. The vectors containing nucleic acid molecules can have one or more expression elements that can be operably connected to such nucleic acid molecules, and can also include sequences such as those encoding selectable markers (for example, antibiotic resistance genes), and / or those sequences that can be used for polypeptide purification (for example, 6xHis tags). Expression elements include nucleic acid sequences that guide and regulate the expression of nucleic acid coding sequences. An example of expression elements is a promoter sequence. Expression elements can also include one or more of the following: introns, enhancer sequences, response elements, or inducible elements that regulate the expression of nucleic acid molecules. Expression elements can be bacteria, yeast, insects, mammals, or viral origins, and vectors can include combinations of expression elements from different sources. As used herein, operably connected means that relative to the coding sequence, expression elements are placed in the vector in the following manner, thereby guiding or regulating the expression of the coding sequence.

[0124] Nucleic acid molecules, for example, nucleic acid molecules in vectors (for example, expression vectors, viral vectors) can be imported into host cells.Term " host cell " refers not only to the specific cells that have accepted nucleic acid molecules to import, but also to the offspring or potential offspring of such cells.Many suitable host cells are well known to those skilled in the art; Host cells can be prokaryotic cells (for example, Escherichia coli) or eukaryotic cells (for example, yeast cells, insect cells, plant cells, mammalian cells).Representational host cells can include but are not limited to A549, WEHI, 3T3, 10T1 / 2, BHK, MDCK, COS 1, COS 7, BSC 1, BSC 40, BMT 10, VERO, WI38, HeLa, 293cells, Saos, C2C12, L cells, HT1080, HepG2 and primary fibroblasts, hepatocytes and myoblast cells derived from mammals (including people, monkey, mouse, rat, rabbit and hamster). Methods for introducing nucleic acid molecules into host cells are well known in the art and include, but are not limited to, calcium phosphate precipitation, electroporation, heat shock, lipofection, microinjection, and virus-mediated nucleic acid transfer (e.g., transduction).

[0125] With respect to polypeptides, "purified" refers to a polypeptide (i.e., a peptide or polypeptide) that has been separated or purified from the cellular components with which it naturally accompanies. Typically, a polypeptide is considered "purified" when it is at least 70% (e.g., at least 75%, 80%, 85%, 90%, 95%, or 99%) free by weight from polypeptides and naturally occurring molecules with which it is naturally associated. Because chemically synthesized polypeptides are inherently separated from the components that naturally accompany them, synthetic polypeptides are considered "purified," but they can also be removed from the components (e.g., amino acid residues) used to synthesize the polypeptide. With respect to nucleic acid molecules, "isolated" refers to a nucleic acid molecule that is separated from other nucleic acid molecules with which it is normally associated in the genome. In addition, isolated nucleic acid molecules can include engineered nucleic acid molecules, such as recombinant or synthetic nucleic acid molecules.

[0126] Polypeptides can be obtained (e.g., purified) from natural sources (e.g., biological samples) by known methods such as DEAE ion exchange, gel filtration, and / or hydroxyapatite chromatography. Purified polypeptides can also be obtained, for example, by expressing nucleic acid molecules in an expression vector or by chemical synthesis. Any suitable method can be used, for example, column chromatography, polyacrylamide gel electrophoresis, or HPLC analysis to measure the purity of the polypeptide. Similarly, conventional methods such as, but not limited to, recombinant nucleic acid technology (e.g., restriction enzyme digestion and connection) or polymerase chain reaction (PCR; See, for example, PCR Primer: A Laboratory Manual, Dieffenbach & Dveksler, Eds., Cold Spring Harbor Laboratory Press, 1995) can be used to obtain (e.g., isolate) nucleic acid molecules. In addition, isolated nucleic acid molecules can be chemically synthesized.

[0127] Methods using ancestral viruses or portions thereof

[0128] Ancestral virus as described herein or its part, particularly those ancestral viruses or its part that show the seroprevalence of reduction relative to contemporary virus or its part, can be used in many research and / or therapeutic applications.For example, ancestral virus as described herein or its part can be used in human or animal medicine for gene therapy (for example, for the carrier or carrier system of gene transfer) or for vaccination (for example, for antigen presentation).More specifically, ancestral virus as described herein or its part can be used for gene addition, gene amplification (augmentation), the genetic delivery of polypeptide therapeutic agent, genetic vaccination, gene silencing, genome editing, gene therapy, RNAi delivery, cDNA delivery, mRNA delivery, miRNA delivery, miRNA sponge wiping (miRNA sponging), genetic immunity, optogenetic gene therapy (optogenetic gene therapy), transgenic effect (transgenesis), DNA vaccination or DNA immunity.

[0129] Host cells can be transduced or infected with the ancestral virus or a portion thereof in vitro (e.g., grown in culture) or in vivo (e.g., in a subject). Host cells that can be transduced or infected with the ancestral virus or a portion thereof in vitro are described herein; host cells that can be transduced or infected with the ancestral virus or a portion thereof in vivo include, but are not limited to, the brain, liver, muscle, lung, eye (e.g., retina, retinal pigment epithelium), kidney, heart, gonads (e.g., testis, uterus, ovary), skin, nasal passages, digestive system, pancreas, islet cells, neurons, lymphocytes, ear (e.g., inner ear), hair follicles and / or glands (e.g., thyroid).

[0130] The ancestral viruses or portions as described herein can be modified to include transgenes (in cis or trans with other viral sequences). The transgene can be, for example, a reporter gene (e.g., β-lactamase, β-galactosidase (LacZ), alkaline phosphatase, thymidine kinase, green fluorescent polypeptide (GFP), chloramphenicol acetyltransferase (CAT), or luciferase, or a fusion polypeptide comprising an antigen tag domain such as hemagglutinin or Myc) or a therapeutic gene (e.g., a gene encoding a hormone or its receptor, a growth factor or its receptor, a differentiation factor or its receptor, an immune system modulator (e.g., a cytokine and interleukin) or its receptor, an enzyme, an RNA (e.g., an inhibitory RNA or a catalytic RNA), or a target antigen (e.g., an oncogenic antigen, an autoimmune antigen)).

[0131] The specific transgene will depend, at least in part, on the specific disease or defect being treated. By way of example only, gene transfer or gene therapy can be applied to the treatment of the following diseases: hemophilia, retinitis pigmentosa, cystic fibrosis, Leber congenital amaurosis, lysosomal storage disorders, inborn errors of metabolism (e.g., inborn errors of amino acid metabolism, including phenylketonuria, inborn errors of organic acid metabolism, including propionic academia, inborn errors of fatty acid metabolism, including medium-chain acyl-CoA dehydrogenase deficiency,MCAD), cancer, achromatopsia, cone-rod dystrophies, macular degeneration (e.g., age-related macular degeneration), lipopolypeptide lipase deficiency, familial hypercholesterolemia, spinal muscular atrophy, Duchenne muscular dystrophy, Alzheimer's disease, Parkinson's disease, obesity, inflammatory bowel disorder, diabetes, congestive heart failure, hypercholesterolemia, hearing loss, coronary heart disease, familial renal amyloidosis, Marfan's syndrome, fatal familial insomnia, Creutzfeldt-Jakob disease, sickle-cell disease disease, Huntington's disease, fronto-temporal lobar degeneration, Usher syndrome, lactose intolerance, lipid storage disorders (such as Niemann-Pick disease type C), Batten disease, choroideremia, glycogen storage disease type II (Pompedisease), ataxia telangiectasia (Louis-Bar syndrome), congenital hypothyroidism, severe combined immunodeficiency (SCID), and / or amyotrophic lateral sclerosis (ALS).

[0132] The transgene can also be an immunogen, for example, that can be used to immunize a subject (e.g., a human, an animal (e.g., a companion animal, a farm animal, an endangered animal). For example, the immunogen can be obtained from an organism (e.g., a pathogenic organism) or an immunogenic portion or component thereof (e.g., a toxin polypeptide or a by-product thereof). By way of example, pathogenic organisms from which immunogenic polypeptides can be obtained include viruses (e.g., picornaviruses, enteroviruses, orthomyxoviruses, reoviruses, retroviruses), prokaryotes (e.g., pneumococci, staphylococci, listeria, pseudomonas), and eukaryotes (e.g., amebiasis, malaria, leishmaniasis, nematodes). It will be understood that the methods described herein and the compositions produced by such methods are not limited to any particular transgene.

[0133] The ancestral virus or its portion, typically suspended in a physiologically compatible carrier, can be administered to a subject (e.g., a human or non-human mammal). Suitable carriers include saline (which can be prepared with a variety of buffer solutions (e.g., phosphate-buffered saline), lactose, sucrose, calcium phosphate, gelatin, dextran, agar, pectin, and water. The ancestral virus or its portion is administered in sufficient amounts to transduce or infect cells and provide sufficient levels of gene transfer and expression to provide therapeutic benefits without undue side effects. Conventional and pharmaceutically acceptable routes of administration include, but are not limited to, direct delivery to organs, such as, for example, the liver or lungs, orally, intranasally, intratracheally, by inhalation, intravenously, intramuscularly, intraocularly, subcutaneously, intradermally, across mucosa, or by other routes of administration. Routes of administration can be combined as needed.

[0134] The dosage of the ancestral virus or portion thereof administered to a subject will depend primarily on factors such as the condition being treated, and the age, weight, and health of the subject. For example, a therapeutically effective dosage of the ancestral virus or portion thereof to be administered to a human subject will generally be in the range of from about 0.1 ml to about 10 ml of a solution containing about 1×10 1 to 1×10 12 The concentration of genome copies (GCS) of the ancestral virus (e.g., about 1 × 10 3 to 1×10 9 GCS). Transduction and / or expression of the transgene can be monitored at multiple time points after administration by DNA, RNA, or protein assays. In some cases, the level of transgene expression can be monitored to determine the frequency and / or amount of dosing. Dosage regimens similar to those described for therapeutic purposes can also be used for immunization.

[0135] The methods described herein can also be used to model forward evolution to modify or remove one or more immunogenic domains of a virus or portion thereof.

[0136] According to the present invention, conventional molecular biology, microbiology, biochemistry and recombinant DNA techniques within the technical scope of this area can be adopted. These technologies are fully explained in the literature. The present invention will be further described in the following examples, which do not limit the scope of the method and composition described in the claims.

[0137] Example

[0138] Example 1: Computer prediction of ancestral sequences

[0139] A set of 75 different amino acid sequences of AAV capsids were obtained from a number of public databases (including GenBank) and the sequences were aligned using the PRANK-MSA algorithm, version 121002, with option "-F".

[0140] ProtTest3 (see, e.g., Darriba et al., 2011, Bioinformatics, 27(8): 1164-5; available on the World Wide Web at darwin.uvigo.es / software / prottest3) was used to evaluate different models of polypeptide evolution (e.g., those included in ProTest3, i.e., JTT, LG, WAG, VT, CpRev, RtRev, Dayhoff, DCMut, FLU, Blosum62, VT, HIVb, MtArt, MtMam) under different conditions (e.g., those included in ProTest3, i.e., "+I", "+F", "+G" and combinations thereof). The JTT model (Jones et al., 1992, Comp. Appl. Biosci., 8:275-82) and +G and +F (Yang, 1993, Mol. Biol. Evol., 10:1396-1401; and Cao et al., 1994, J. Mol. Evol., 39:519-27) were selected based on the Aikake information criterion (AIC; Hirotugu, 1974, IEEE Transactions on Automatic Control, 19:716-23) scores as implemented in ProTest3.

[0141] PhyML (Guindon and Gascuel, 2003, Systematic Biology, 52: 696-704) was used to construct a phylogeny of AAV evolution. Figure 3 The tree was generated using the JTT+F substitution model with four discrete substitution classes and an estimated gamma shape parameter. The resulting tree was improved by nearest neighbor interchange (NNI) and subtree pruning and replanting (SPR), and significance was assessed using the "SH-Like" variant using the bootstrap method and the approximate likelihood ratio test (aLRT; Anisimova and Gascuel, 2006, Systematic Biology, 55:539-52).

[0142] The phylogenetic tree constructed above was then used to estimate the ancestral state of the AAV capsid at each node within the phylogeny. Ancestral capsid sequences were reconstructed using the maximum likelihood principle using the Phylogenetic Analysis by Maximum Likelihood (PAML) software (Yang, 1997, Comp. Applic. BioSci., 13:555-6; available on the World Wide Web at abacus.gene.ucl.ac.uk / software / paml.html) included in Lazarus (Sourceforge at sf.net). More specifically, Lazarus / PAML reconstructions were set up to generate amino acid reconstructions using a JTT+F substitution model using a 4-gamma distribution class. AAV5 was used as an outgroup. Finally, the "I" option was added to place indels after the PAML reconstruction was completed (i.e., binary coded and placed using the Fitch algorithm by Maximum Parsimony).

[0143] Because the reconstruction is performed in a maximum likelihood manner, the likelihood of any residue at a given position at a given node can be calculated. To do this, additional scripts were written to identify all positions along the sequence with a calculated posterior probability below a certain threshold. A threshold of 0.3 was chosen, meaning that any amino acid with a calculated posterior probability greater than 0.3 was included in the synthesis of the library. These residues were selected as variants of interest in the library.

[0144] To finalize the sequence, additional utilities had to be coded for codon selection. A script was written to derive codons similar to those in another AAV sequence (AVVRh10, which has approximately 92% sequence identity to the Anc80 scaffold sequence) and apply the new algorithm to replace codons where there were sequence mismatches based on the codon substitution matrix. The new algorithm is shown below:

[0145] Given: an amino acid sequence, Pt, with a corresponding nucleotide sequence, NT, where Nt encodes Pt; and a protein sequence, Pi, where Pi exhibits strong homology to Pt.

[0146] Pi was aligned to Pt using Needleman-Wunsch using the Blosum62 table for scoring. A new nucleotide sequence, Ni, was generated by stepping through the protein alignment using the corresponding codons from Nt.

[0147] The amino acids in Pt completely match those in Pi.

[0148] The "best scoring" codon from the Codon-PAM matrix (Schneider et al., 2005, BMC Bioinform., 6:134)) where the substitution occurred,

[0149] gaps, where there are gaps in Pi that align with the amino acids in Pt, and

[0150] The most frequently occurring nucleotide (encoding a given amino acid) in Nt where the amino acid is present in Pi aligned to a gap in Pt.

[0151] In addition, two single nucleotide changes were made to eliminate transcription of the assembly-activating protein (AAP), which is encoded out-of-frame within the AAV capsid gene in wild-type AAV. Because the coding for AAP (contemporary or ancestral) was not part of this reconstruction, AAP expression was eliminated by generating synonymous mutations in the cap sequence, and the AAP sequence was provided in trans during virus production.

[0152] Example 2: Expression of ancestral AAV VP1 sequences

[0153] Experiments were performed to determine whether the predicted ancestral AAV capsid sequence could be used to prepare viral vectors.

[0154] A number of predicted ancestral AAV capsid sequences were cloned. The ancestral capsid library was transferred to the rep-cap expression plasmid, enabling viral particle formation in transient transfections. To maintain appropriate expression levels and splicing of VP1, VP2, and VP3, the library cap gene was cloned by HindIII, which cuts 5' of the cap in the rep coding sequence, and SpeI, which was engineered between the cap stop codon and the polyadenylation signal. Therefore, to clone the ancestral capsid into the more conventional "REP / CAP" construct, the passage plasmid was digested with HindIII and SpeI, gel purified, and ligated into a similarly digested rep / cap plasmid.

[0155] The expressed polypeptides were resolved on 10% SDS gel. Figure 6 As shown in , capsid polypeptides are appropriately expressed and spliced ​​into VP1, VP2, and VP3 from multiple ancestral AAV sequences (Anc80L44, Anc80L27, and Anc80L65) as well as from contemporary AAV sequences, AAV2 / 8.

[0156] Example 3: Virus titration

[0157] AAV was produced in HEK293 cells by transient co-transfection of plasmids encoding all the elements required for viral particle assembly. Briefly, HEK293 cells were cultured to 90% confluence and transfected with: (a) a viral genomic plasmid encoding the luciferase transgene (expressed by the CMV promoter) flanked by AAV2 ITRs, (b) an AAV packaging plasmid encoding the AAV2 rep and synthetic capsid proteins disclosed herein, (c) an AAV2-AAP expression capsid, and (d) adenoviral helper genes required for AAV packaging and assembly. Cells were cultured at 37°C for 2 days, and the cells and culture medium were harvested and collected.

[0158] The cell culture suspension was lysed by three consecutive freeze-thaw cycles. The lysate was then clarified by centrifugation and treated with an enzyme (Benzonase in this case) under conditions that allowed for thorough DNA digestion. TM ) to digest any DNA present outside the viral particles. The AAV preparation was diluted to fall within the linear measurement range of the control DNA template, in this case, the same TaqMan TM TaqMan PCR is performed using primers and probes that anneal to the viral vector genome of choice. TM PCR. TaqMan-based TM Titers were calculated as genome copies (GC) per milliliter (ml) as shown in Table 2 below.

[0159] Table 2

[0160] Titer (GC / ml) Small Scale #1 Small Scale #2 AAV2 / 2 <![CDATA[1.12x10 9 ]]> <![CDATA[1.99x10 9 ]]> AAV2 / 8 <![CDATA[4.17x10 10 ]]> <![CDATA[5.91x10 10 ]]> Anc80L27 <![CDATA[8.01x10 8 ]]> <![CDATA[1.74x10 9 ]]> Anc80L44 <![CDATA[1.52x10 9 ]]> <![CDATA[1.43x10 9 ]]> Anc80L65 <![CDATA[1.42x10 9 ]]> <![CDATA[2.05x10 9 ]]> No shell control <![CDATA[5.23x10 5 ]]> <![CDATA[7.25x10 5 ]]>

[0161] Small-scale vector production results on ancestral reconstituted AAV capsid particles demonstrated yields similar to AAV2, but reduced relative to AAV8 (both of which are contemporary AAV-based vector preparations).

[0162] Example 4: In vitro viral transduction

[0163] In vitro viral transduction was performed to evaluate the ability of viruses containing predicted ancestral AAV sequences to infect cells.

[0164] After high-throughput vector production using the Anc80 sequence library, HEK293 cells were transduced with each viral vector. In addition to the Anc80 sequence, each viral vector contained a luciferase transgene. After adding the luciferin substrate to the transduced cells or cell lysates, luciferase was measured by quantitative bioluminescence in a 96-well plate reader. After quantification, a heat map of luciferase expression in four cascaded 96-well plates was generated (excluding the control column in each plate). Due to the large number of insertions, deletions, and conversions associated with the high-throughput vector production process, many vectors are non-functional. For the purposes of this article, only functional viruses in this assay (i.e., capable of transducing HEK293 cells and expressing transgenes) were further evaluated.

[0165] Two contemporary AAV vectors (AAV2 / 2 and AAV2 / 8) and three predicted ancestral AAV vectors (Anc80L27, Anc80L44, and Anc80L65) were used at 1 × 10 per cell. 4 HEK293 cells were transduced with equal multiplicities of infection (MOI) of 100 genomic copies (GC). Each vector contained either a luciferase-encoding transgene or an eGFP-encoding transgene. Cells were imaged 60 hours later using the GFP channel of an AMGEvosF1 light microscope. Figure 7 Luciferase expression after in vitro transduction is shown. Each progenitor AAV virus demonstrated efficient transduction of HEK293 cells.

[0166] Example 5 - In vivo retinal transduction

[0167] Retinal transduction was performed to determine whether the ancestral AAV vector could target murine retinal cells in vivo.

[0168] Use 2×10 8Three different ancestral AAVs (Anc80L27, Anc80L44, and Anc80L65) and contemporary AAVs (AAV2 / 8) of the genomic copy (GC) were transduced into mouse eyes, all of which included the eGFP encoding transgene. For transduction, each AAV vector was surgically delivered beneath the retina by creating a space between the photoreceptors and the retinal pigment epithelium via a vector bolus delivered with an injection device. The vector bolus remained in the subretinal space and resolved retinal detachment over time. TM Fundus photography of the retinas of animals with pupils dilated was performed to non-invasively monitor GFP expression. All retinas presented demonstrated varying degrees of successful targeting of the progenitor AAV to the retina.

[0169] Retinal histology was also performed and visualized under fluorescence microscopy to identify transduced cell types. Histology was performed on mouse retinas transduced with the Anc80L65 ancestral AAV vector as described above. Anc80L65-mediated eGFP expression was evident in the outer nuclear layer (ONL), inner segments (IS), and retinal pigment epithelium (RPE), indicating that the ancestral Anc80L65 vector targets mouse photoreceptors and retinal pigment epithelial cells.

[0170] Example 6: Neutralizing Antibody Assay

[0171] Neutralizing antibody assays were performed to assess whether ancestral AAV viruses are more resistant to antibody neutralization than contemporary AAV viruses. Neutralizing antibody assays measure the concentration of antibodies (or the titer of antibody concentration in experimental samples) that neutralize infection by 50% or more compared to a control in the absence of antibody.

[0172] Serum samples or IVIG stock solution (200 mg / ml) were serially diluted 2-fold, and the undiluted and diluted samples were mixed with 10 4 The ancestral AAV virus, Anc80L65, and the contemporary AAV virus, AAV2 / 8, were co-incubated at 37°C for approximately 30 minutes at an MOI of 1:1. Each virus included a luciferase gene. The mixed vector and antibody samples were then transduced into HEK293 cells. For these experiments, the antibody sample used was intravenous immunoglobulin (IVIG) obtained from over 1000 blood donors (commercially available, e.g., Gammagard TM (Baxter Healthcare; Deerfield, IL) or Gamunex TMPooled IgG was extracted from plasma of 184 healthy volunteers (Grifols; Los Angeles, CA). 48 hours after initiation of transduction, cells were assayed by bioluminescence to detect luciferase. Neutralizing antibody titers were determined by identifying the dilution of sample that achieved 50% or greater neutralization (sample transduction / transduction of control virus in the absence of sample).

[0173] like Figure 8 As shown, to reduce the transduction efficiency of the ancestral AAV virus Anc80L65 to below 50% of the no IVIG control (dashed line), significantly higher concentrations of IVIG were required compared to contemporary AAV viruses AAV2 / 8. These results indicate that ancestral AAV viruses are more resistant to neutralization by IVIG than contemporary AAV viruses.

[0174] Example 7: Characterization of Anc80

[0175] According to the methods described herein, the most likely Anc80 sequence (as determined by posterior probability) was obtained and designated Anc80L1 (SEQ ID NO: 35 shows the nucleic acid sequence of the Anc80L1 capsid and SEQ ID NO: 36 shows the amino acid sequence of the Anc80L1 VP1 polypeptide). Anc80 probabilistic libraries were also synthesized by commercial companies using the sequences described herein and subcloned into expression vectors.

[0176] The Anc80 library was cloned and evaluated for vector yield and infectivity in a combination assay. In the screen, Anc80L65 (SEQ ID NO: 23) and several other variants were further characterized.

[0177] The Anc80 library and Anc80L65 ( Figure 9 % up from the diagonal, # of amino acid differences below). Using NCBI-BLAST, the closest publicly available sequence to Anc80L65 is rh10 (GenBank Accession No. AAO88201.1).

[0178] Figure 10 Anc80L65 was shown to produce vector yields comparable to AAV2 (Panel A), viral particles under transmission electron microscopy (TEM) (Panel B), and biochemically produced AAV cap and VP1, 2, and 3 proteins based on SDS-PAGE under denaturing conditions (Panel C) and Western blotting using AAV capsid antibody B1 (Panel D). These experiments are described in more detail in the following paragraphs.

[0179] Briefly, AAV2 / 8, AAV2 / 2, AAV2 / Anc80L27, AAV2 / Anc80L44, and AAV2 / Anc80L65 vectors containing reporter constructs consisting of eGFP and firefly luciferase under the control of the CMV promoter were produced on a small scale. The titers of these small-scale preparations of virus were then obtained by qPCR. Based on these experiments, it was found that the Anc80L27, Anc80L44, and Anc80L65 vectors produced virus levels comparable to those of AAV2 ( Figure 10 A).

[0180] To confirm that the Anc80L65 capsid protein assembled into complete virus-like particles of appropriate size and conformation, micrographs were obtained using transmission electron microscopy (TEM). The coated copper grid was then stained with uranyl acetate. Micrographs revealed intact, hexagonal particles with diameters between 20 and 25 nm ( Figure 10 B).

[0181] To determine whether the synthesized ancestral capsid genes were properly processed (i.e., spliced ​​and expressed), large-scale purified preparations of AAV2 / 8, AAV2 / 2, and AAV2 / Anc80L65 vectors were loaded onto SDS-PAGE gels (1E10 GC / well) under denaturing conditions. For each vector preparation, bands representing viral capsid proteins VP1, VP2, and VP3 were clearly present ( Figure 10 C). Western blotting using AAV capsid antibody B1 further confirmed that these bands represent the predicted proteins ( Figure 10 D).

[0182] also, Figure 11 Anc80L65 was shown to infect mammalian tissues and cells in vitro on HEK293 cells at an MOI of 10E4 GC / cell relative to AAV2 and / or AAV8 controls using GFP as a readout (Panel A) or luciferase (Panel B). Anc80L65 was also effectively targeted to the liver following IV injection of the indicated AAV encoding the nuclear LacZ transgene (top row, Panel C), direct intramuscular (IM) injection of the indicated AAV encoding GFP (middle row, Panel C), and subretinal injection of the indicated AAV encoding GFP (bottom row, Panel C). These experiments are described in more detail in the following paragraphs.

[0183] To obtain a relative measure of the infectivity of ancestral virions, crude preparations of AAV2 / 2, AAV2 / 8, AAV2 / Anc80L65, AAV2 / Anc80L44, AAV2 / Anc80L27, AAV2 / Anc80L121, AAV2 / Anc80L122, AAV2 / Anc80L123, AAV2 / Anc80L124, and AAV2 / Anc80L125 containing bicistronic reporter constructs were generated, including eGFP and firefly luciferase under the control of the CMV promoter. Each vector was then used to transduce confluent 96-well plates of HEK293 cells at an MOI of 1E4 GC / cell (titer obtained by qPCR as described above). After 48 hours, the presence of GFP in the transduced cells was confirmed by fluorescence microscopy ( Figure 11 The cells were then assayed for the presence of luciferase ( Figure 11 B), which determined that luciferase expression in cells transduced with Anc80-derived vectors was intermediate between that in cells transduced with AAV8 (lower level of transduction) and in cells transduced with AAV2 (higher level of transduction).

[0184] To assess the relative efficiency of gene transfer in vivo, purified high-titer preparations of AAV2 / 2, AAV2 / 8, and AAV2 / Anc80L65 were obtained. 3.9E10 GC of each vector (encapsidating a transgene encoding nuclear LacZ under the control of the TBG promoter) was injected into C57BL / 6 mice (3 mice per condition) by IP injection after general anesthesia. 28 days after injection, mice were sacrificed and tissues were collected. Liver sections were sectioned by standard histological techniques and stained for β-galactosidase. Sections were then imaged under a microscope, and representative images are shown in Figure 2. Figure 11 C, top row.

[0185] A vector of the same serotype was then obtained containing a bicistronic transgene encoding eGFP and hA1AT under the control of the pCASI promoter. To evaluate the ability of Anc80L65 to transduce murine skeletal muscle, 1E10 GC of each vector was injected into the skeletal muscle of C57BL / 6 mice under general anesthesia (5 mice per condition). 28 days after injection, the mice were sacrificed, tissues were frozen, and the presence of eGFP was assessed by fluorescence confocal microscopy (blue is DAPI, green is EGFP). Representative images are shown in Figure 11 C, middle row. These experiments demonstrate that the Anc80L65 vector is capable of transducing murine skeletal muscle by intramuscular injection.

[0186] A vector of the same serotype was obtained, this time encapsidating a construct encoding only the eGFP transgene under the control of the CMV promoter. 2E9 particles were injected sub-retinally into C57BL / 6 mice after general anesthesia. 28 days after injection, mice were sacrificed, eyes were harvested, frozen, and the presence of eGFP was assessed using fluorescence confocal microscopy (blue is DAPI, green is EGFP). Representative images are shown in Figure 11 C, bottom row. These experiments demonstrate that the Anc80L65 vector is able to transduce the mouse retina at levels comparable to those of the AAV8 vector.

[0187] Figure 12 Results of experiments are presented in which the seroprevalence of ancestral viral vectors was assessed relative to existing AAV viral vectors. Using an in vitro neutralizing antibody assay, Anc80L65 was shown to exhibit increased resistance to neutralization using IVIG (medicinal pooled sera from approximately 10,000 individuals) ( Figure 12 In addition, in the whisker box plot, sera from Belgian individuals (n = 100; Figure 12 C) or sera from Boston individuals (n = 102; Figure 12 B) shows reduced sensitivity (or increased resistance) to neutralization of Anc80L65 relative to AAV2,8, and similar seroprevalence relative to rh32.33 (a different AV vector with the lowest known seroprevalence but limited use as a gene therapy vector). Sera obtained from cynomolgus macaques exhibited similar increased resistance ( Figure 12 D). These experiments are described in more detail in the following paragraphs.

[0188] In brief, purified high-titer preparations of AAV2 / 8, AAV2 / 2, AAV2 / rh32.33 and AAV2 / Anc80L65 viral vectors were obtained, and the viral vector encapsidation included eGFP and firefly luciferase bicistronic transgenes under the control of the CMV promoter. These vectors were then incubated with or without IVIG (1E9 GC per condition) with two-fold serial dilutions of IVIG (10 mg, 5 mg, 2.5 mg, etc.). After incubation, the vector was used to transduce HEK293 cells at an MOI of 1E4 per well (one dilution per well). After 48 hours, the relative amount of luciferase was determined by luminescence assay. Transduction relative to serum-free control is shown in Figure 12 A. Using sera from Belgian individuals (n = 100; Figure 12 C) or sera from Boston individuals (n = 102; Figure 12A similar experiment was performed for B). For each serum sample, the neutralization titer is reported as the dilution at which a given vector is reduced by 50% relative to the serum-free control. Figure 12 Neutralization titers are reported in box-and-whisker plots in Figures B and C, which confirm that the Anc80L65 vector has a lower prevalence in both populations (Belgian or Boston) than AAV2 and AAV8, approaching the level of Rh32.33 (the different AAV vectors with the lowest known seroprevalence). Sera obtained from cynomolgus macaques ( Figure 12 D), where Anc80L65 was found to have a significantly lower seropositivity rate compared to AAV2, AAV8, and Rh.32.33 vectors.

[0189] Example 8: Generation of other ancestral AAV capsids

[0190] The most likely ancestral AAV capsid sequences (as determined by posterior probability) were then synthesized by a commercial laboratory (Gen9) and provided as linear dsDNA. These amino acid sequences were then compared with the amino acid sequences of existing AAVs to determine the extent to which they differed ( FIG13 ). Each ancestral VP1 protein differed from the VP1 protein of a selected representative existing AAV by 3.6% to 9.3% ( Figure 13A ), while the ancestral VP3 protein differed by 4.2%-9.4% ( Figure 13B Each of these capsids was subcloned into an AAV production plasmid (pAAVector2 / empty) by restriction enzyme digestion (HindIII & SpeI) and T4 ligation. These clones were confirmed by restriction digestion and Sanger sequencing, and then a medium-scale plasmid DNA preparation was generated.

[0191] Each of these plasmids was then used to generate AAV vectors containing reporter genes encoding both eGFP and firefly luciferase. These vectors were produced in triplicate on a small scale as described previously. Crude preparations of virus were then titrated by qPCR and found to produce 2.71% to 183.1% more viral particles relative to AAV8 ( Figure 14 and 15 These titers were then used to set up titer control experiments to assess relative infectivity. No titer control was available for Anc126 due to its significantly reduced production; therefore, data regarding the infectivity of Anc126 cannot be accurately compared to the infectivity of the other viruses in the experiment. The other vectors were used to transduce HEK293 cells at a multiplicity of infection (MOI) of 1.9 E3 GC / cell.

[0192] 60 hours after transduction, cells were evaluated for GFP expression by fluorescence microscopy. In addition to the negative control, eGFP-positive cells ( Figure 16This indicates that each of the predicted, synthesized, and cloned ancestral sequences was able to produce viable, infectious viral particles. To obtain an idea of ​​the relative levels of infectivity, luciferase assays were also performed on the same cells. The results indicate that each ancestral vector was able to transduce 28.3% to 850.8% of HEK293 cells relative to AAV8 ( Figure 17 and 18 ). It should be noted that Anc126 was excluded from the analysis of relative transduction because it had no titer control.

[0193] In summary, eight new ancestral AAV capsid genes were synthesized and used with AAV8, AAV2, and the previously described Anc80L65 vectors to generate functional viral vectors. Production and infection were assessed in vitro, and a summary of those findings is shown in Figure 19 .

[0194] Example 9: Vector Immunization Prevention

[0195] In vectored immunoprophylaxis, gene therapy vectors (such as AAV) are used to deliver transgenes encoding broadly neutralizing antibodies against infectious agents. See, for example, Balazs et al. (2013, Nat. Biotechnol., 31: 647-52); Limberis et al. (2013, Sci. Transl. Med., 5: 187ra72); Balazs et al. (2012, Nature, 481: 81-4); and Deal et al. (2014, PNAS USA, 111: 12528-32). One advantage of this treatment is that the host produces antibodies in their own cells, which means that a single administration has the potential to confer lifelong protection against etiologic agents.

[0196] Example 10: Drug delivery vehicle

[0197] (ranibizumab) and Avastin (bevacizumab) are both anti-angiogenic agents based on the same humanized mouse monoclonal antibody against vascular endothelial growth factor A (VEGF-A). Although bevacizumab is a complete antibody and ranibizumab is a fragment (Fab), they act through the same mechanism - by antagonizing VEGF to treat wet age-related macular degeneration. See, for example, Mao et al. (2011, Hum. Gene Ther., 22: 1525-35); Xie et al. (2014, Gynecol. Oncol., doi: 10.1016 / j.ygyno.2014.07.105); and Watanabe et al. (2010, Gene Ther., 17: 1042-51). Because both molecules are proteins, they can be encoded by DNA and produced in cells transduced with a vector containing a transgene, and are small enough to be packaged into an AAV vector.

[0198] Other implementation plans

[0199] It should be understood that although the method and material composition have been described herein in conjunction with various aspects, the description of the aforementioned aspects is intended to illustrate rather than limit the scope of the method and material composition. Other aspects, advantages, and modifications are within the scope of the appended claims.

[0200] Disclosed herein are methods and compositions that can be used for the products of the disclosed methods and compositions, can be used in conjunction with the products of the disclosed methods and compositions, can be used to prepare the products of the disclosed methods and compositions or are products of the disclosed methods and compositions. These and other substances are disclosed herein, and it should be understood that combinations, subgroups, interactions, groups, etc. of these methods and compositions are disclosed. That is, although each different individual and collective combination and replacement of these compositions and methods may not be explicitly disclosed and specifically mentioned, each is specifically contemplated and described herein. For example, if a specific composition of matter or ad hoc method is disclosed and discussed and multiple compositions and methods are discussed, then unless there is a clear indication to the contrary, each combination and replacement of compositions and methods is explicitly covered. Likewise, any subgroup or combination of these is also explicitly covered and disclosed.

[0201] Annex A

[0202] SEQ ID NO: 1: Anc80 polypeptide

[0203]

[0204] SEQ ID NO: 2: Anc80 DNA

[0205]

[0206]

[0207] SEQ ID NO: 3: Anc81 polypeptide

[0208] SEQ ID NO: 4: Anc81 DNA

[0209]

[0210]

[0211] SEQ ID NO: 5: Anc82 polypeptide

[0212]

[0213] SEQ ID NO: 6: Anc82 DNA

[0214]

[0215]

[0216] SEQ ID NO: 7: Anc83 polypeptide

[0217]

[0218] SEQ ID NO: 8: Anc83 DNA

[0219]

[0220]

[0221] SEQ ID NO: 9: Anc84 polypeptide

[0222]

[0223] SEQ ID NO: 10: Anc84 DNA

[0224]

[0225]

[0226] SEQ ID NO: 11: Anc94 polypeptide

[0227]

[0228] SEQ ID NO: 12: Anc94 DNA

[0229]

[0230]

[0231] SEQ ID NO: 13: Anc113 polypeptide

[0232] SEQ ID NO: 14: Anc113 DNA

[0233]

[0234]

[0235] SEQ ID NO: 15: Anc126 polypeptide

[0236]

[0237] SEQ ID NO: 16: Anc126 DNA

[0238]

[0239]

[0240] SEQ ID NO: 17: Anc127 polypeptide

[0241]

[0242] SEQ ID NO: 18: Anc127 DNA

[0243]

[0244]

[0245] SEQ ID NO: 19: L0027 polypeptide

[0246]

[0247] SEQ ID NO: 20: L0059 polypeptide

[0248]

[0249] SEQ ID NO: 21: L0060 polypeptide

[0250]

[0251] SEQ ID NO: 22: L0062 polypeptide

[0252]

[0253]

[0254] SEQ ID NO: 23: L0065 polypeptide

[0255]

[0256] SEQ ID NO: 24: L0033 polypeptide

[0257]

[0258] SEQ ID NO: 25: L0036 polypeptide

[0259]

[0260] SEQ ID NO: 26: L0044 polypeptide

[0261]

[0262]

[0263] SEQ ID NO: 27: AAV8 VP1 polypeptide

[0264] SEQ ID NO: 28: AAV9 VP1 polypeptide

[0265] SEQ ID NO: 29: AAV6 VP1 polypeptide

[0266] SEQ ID NO: 30: AAV1 VP1 polypeptide

[0267] SEQ ID NO: 31: AAV2 VP1 polypeptide

[0268] SEQ ID No: 32: AAV3 VP1 polypeptide

[0269] SEQ ID No: 33: AAV3B VP1 polypeptide

[0270] SEQ ID NO: 34: AAV7 VP1 polypeptide Sequence Listing <110> Massachusetts Eye and Ear Clinic <120> Methods for predicting ancestral viral sequences and uses thereof <130> 00633-0155WO1 <140> PCT / US2014 / 060163 <141> 2014-10-10 <150> 61 / 889,827 <151> 2013-10-11 <160> 44 <170> PatentIn Version 3.5 <210> 1 <211> 736 <212> PRT <213> Adeno-associated virus <220> <221> Variants <222> (168)..(168) <223> / Replace="Arg" <220> <221> Variants <222> (204)..(204) <223> / Replace="Ser" <220> <221> Variants <222> (266)..(266) <223> / Replace="Gly" <220> <221> Variants <222> (311)..(311) <223> / replace="Lys" <220> <221> Variants <222> (411)..(411) <223> / Replace="Gln" <220> <221> Variants <222> (460)..(460) <223> / Replace="Glu" <220> <221> Variants <222> (493)..(493) <223> / Replace="Thr" <220> <221> Variants <222> (562)..(562) <223> / Replace="Asn" <220> <221> Variants <222> (576)..(576) <223> / Replace="Glu" <220> <221> Variants <222> (587)..(587) <223> / Replace="Ala" <220> <221> Variants <222> (609)..(609) <223> / Replace="Asp" <220> <221> misc_feature <222> (1)..(736) <223> / note="There is no preference for variant residues given in the sequence relative to those in the annotation at the variant position" <400> 1 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ala Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala Ser Thr Asn Asp Asn Thr 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Thr Phe 405 410 415 Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg 435 440 445 Thr Gln Thr Thr Ser Gly Thr Ala Gly Asn Arg Thr Leu Gln Phe Ser 450 455 460 Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ala Asn Gln Asn 485 490 495 Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Thr His Lys 515 520 525 Asp Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Leu Ile Phe Gly 530 535 540 Lys Gln Gly Ala Gly Asn Ser Asn Val Asp Leu Asp Asn Val Met Ile 545 550 555 560 Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Gln 565 570 575 Tyr Gly Thr Val Ala Thr Asn Leu Gln Ser Ser Asn Thr Ala Pro Ala 580 585 590 Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Thr Asn Val Asp Phe Ala Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 2 <211> 2208 <212> DNA <213> Adeno-associated virus <220> <221> Variant <222> (502)..(504) <223> / replace="aaa" <220> <221> Mutations <222> (610)..(612) <223> / replace="agc" <220> <221> Mutations <222> (796)..(798) <223> / replace="ggc" <220> <221> Mutations <222> (931)..(933) <223> / replace="aag" <220> <221> Mutations <222> (1231)..(1233) <223> / replace="cag" <220> <221> Mutations <222> (1378)..(1380) <223> / replace="gag" <220> <221> Mutations <222> (1477)..(1479) <223> / replace="acc" <220> <221> Mutations <222> (1684)..(1686) <223> / replace="aac" <220> <221> Mutations <222> (1726)..(1728) <223> / replace="gag" <220> <221> Mutations <222> (1759)..(1761) <223> / replace="gcc" <220> <221> Mutations <222> (1825)..(1827) <223> / replace="gac" <220> <221> misc_feature <222> (1)..(2208) <223> / note="There is no preference for variant nucleotides given in the sequence relative to those in the variant position annotation" <400> 2 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acttgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata accacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 ggaaagaaga gaccggtaga gcaatcaccc caggaaccag actcctcttc gggcatcggc 480 aagaaaggcc agcagcccgc gaagaagaga ctcaactttg ggcagacagg cgactcagag 540 tcagtgcccg accctcaacc actcggagaa ccccccgcag ccccctctgg tgtgggatct 600 aatacaatgg cagcaggcgg tggcgctcca atggcagaca ataacgaagg cgccgacgga 660 gtgggtaacg cctcaggaaa ttggcattgc gattccacat ggctgggcga cagagtcatc 720 accaccagca cccgaacctg ggccctcccc acctacaaca accacctcta caagcaaatc 780 tccagccaat cgggagcaag caccaacgac aacacctact tcggctacag caccccctgg 840 gggtattttg actttaacag attccactgc cacttctcac cacgtgactg gcagcgactc 900 atcaacaaca actggggatt ccggcccaag agactcaact tcaagctctt caacatccag 960 gtcaaggagg tcacgacgaa tgatggcacc acgaccatcg ccaataacct taccagcacg 1020 gttcaggtct ttacggactc ggaataccag ctcccgtacg tcctcggctc tgcgcaccag 1080 ggctgcctgc ctccgttccc ggcggacgtc ttcatgattc ctcagtacgg gtacctgact 1140 ctgaacaatg gcagtcaggc cgtgggccgt tcctccttct actgcctgga gtactttcct 1200 tctcaaatgc tgagaacggg caacaacttt gagttcagct acacgtttga ggacgtgcct 1260 tttcacagca gctacgcgca cagccaaagc ctggaccggc tgatgaaccc cctcatcgac 1320 footcctgt actacctgtc tcggactcag accacgagtg gtaccgcagg aaatcggacg 1380 ttgcaatttt ctcaggccgg gcctagtagc atggcgaatc aggccaaaaa ctggctaccc 1440 gggccctgct accggcagca acgcgtctcc aagacagcga atcaaaaataa caacagcaac 1500 tttgcctgga ccggtgccac caagtatcat ctgaatggca gagactctct ggtaaatccc 1560 ggtcccgcta tggcaaccca caaggacgacgac gaacaaat tttttccgat gagcggagtc 1620 ttaatatttg ggaacaggg agctgggaaat agcaacgtgg accttgacaa cgttatgata 1680 accagtgagg aagaaattaa aaccaccaac ccagtggcca cagaacagta cggcacggtg 1740 gccactaacc tgcaatcgtc aaacaccgct cctgctacag ggaccgtcaa cagtcaagga 1800 gccttacctg gcatggtctg gcagaaccgg gacgtgtacc tgcagggtcc tatctgggcc 1860 aagattcctc acacggacgg acactttcat ccctcgccgc tgatgggagg ctttggactg 1920 aaacacccgc ctcctcagat cctgattaag aatacacctg ttcccgcgaa tcctccaact 1980 accttcagtc cagctaagtt tgcgtcgttc atcacgcagt acagcaccgg acaggtcagc 2040 gtggaaattg aatggggagct gcagaaagaa aacagcaaac gctggaaccc agagattcaa 2100 tacacttcca actacaacaa atctacaaat gtggactttg ctgttgacac aaatggcgtt 2160 tattctgagc ctcgccccat cggcacccgt tacctcaccc gtaatctg 2208 <210> 3 <211> 737 <212> PRT <213> Adeno-associated virus <220> <221> Variants <222> (157)..(157) <223> / Replace="Ser" <220> <221> Variants <222> (168)..(168) <223> / Replace="Arg" <220> <221> Variants <222> (262)..(262) <223> / Replace="Ser" <220> <221> Variants <222> (263)..(263) <223> / Replace="His" <220> <221> Variants <222> (312)..(312) <223> / replace="Lys" <220> <221> Variants <222> (412)..(412) <223> / Replace="Gln" <220> <221> Variants <222> (460)..(460) <223> / Replace="Gln" <220> <221> Variants <222> (461)..(461) <223> / Replace="Glu" <220> <221> Variants <222> (552)..(552) <223> / Replace="Ser" <220> <221> Variants <222> (556)..(556) <223> / replace="Tyr" <220> <221> Variants <222> (557)..(557) <223> / Replace="Ser" <220> <221> Variants <222> (563)..(563) <223> / Replace="Asn" <220> <221> Variants <222> (580)..(580) <223> / Replace="Ile" <220> <221> Variants <222> (588)..(588) <223> / Replace="Ser" <220> <221> Variants <222> (664)..(664) <223> / Replace="Thr" <220> <221> misc_feature <222> (1)..(737) <223> / note="There is no preference for variant residues given in the sequence relative to those in the annotation at the variant position" <400> 3 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Thr Gly Ile Gly 145 150 155 160 Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ala Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Asn Ser Gln Ser Gly Gly Ser Thr Asn Asp Asn 260 265 270 Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg 275 280 285 Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn 290 295 300 Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile 305 310 315 320 Gln Val Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn 325 330 335 Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu 340 345 350 Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro 355 360 365 Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn 370 375 380 Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe 385 390 395 400 Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Thr 405 410 415 Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu 420 425 430 Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser 435 440 445 Arg Thr Gln Thr Thr Gly Gly Thr Ala Gly Asn Arg Thr Leu Gln Phe 450 455 460 Ser Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu 465 470 475 480 Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Thr Asn Gln 485 490 495 Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu 500 505 510 Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr His 515 520 525 Lys Asp Asp Glu Asp Arg Phe Phe Pro Ser Ser Gly Val Leu Ile Phe 530 535 540 Gly Lys Gln Gly Ala Gly Asn Asp Asn Val Asp Leu Asp Asn Val Met 545 550 555 560 Ile Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu 565 570 575 Glu Tyr Gly Val Val Ala Thr Asn Leu Gln Ser Ala Asn Thr Ala Pro 580 585 590 Gln Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp 595 600 605 Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro 610 615 620 His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly 625 630 635 640 Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro 645 650 655 Ala Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile 660 665 670 Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu 675 680 685 Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser 690 695 700 Asn Tyr Asn Lys Ser Thr Asn Val Asp Phe Ala Val Asp Thr Glu Gly 705 710 715 720 Val Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn 725 730 735 Leu <210> 4 <211> 2211 <212> DNA <213> Adeno-associated virus <220> <221> Mutations <222> (469)..(471) <223> / replace="agc" <220> <221> Mutations <222> (502)..(504) <223> / replace="aag" <220> <221> Mutations <222> (784)..(786) <223> / replace="agt" <220> <221> Mutations <222> (787)..(789) <223> / replace="cac" <220> <221> Mutations <222> (934)..(936) <223> / replace="aag" <220> <221> Mutations <222> (1234)..(1236) <223> / replace="cag" <220> <221> Mutations <222> (1378)..(1380) <223> / replace="cag" <220> <221> Mutations <222> (1381)..(1383) <223> / replace="gag" <220> <221> Mutations <222> (1654)..(1656) <223> / replace="agc" <220> <221> Mutations <222> (1666)..(1668) <223> / replace="tac" <220> <221> Mutations <222> (1669)..(1671) <223> / replace="agc" <220> <221> Mutations <222> (1687)..(1689) <223> / replace="aac" <220> <221> Mutations <222> (1738)..(1740) <223> / replace="atc" <220> <221> Mutations <222> (1762)..(1764) <223> / replace="agc" <220> <221> Mutations <222> (1990)..(1992) <223> / replace="acc" <220> <221> misc_feature <222> (1)..(2211) <223> / note="There is no preference for variant nucleotides given in the sequence relative to those in the variant position annotation" <400> 4 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acttgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata accacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 ggaaaagaa gaccggtaga gcaatcaccc caggaaccag actcctctac gggcatcgg 480 aagaaaggcc agcagcccgc gaaaagaga ctcaactttg ggcagactgg cgactcagag 540 tcagtgcccg accctcaacc actcggagaa ccccccgcag ccccctctgg tgtgggatct 600 aatacaatgg ctgcaggcgg tggcgctcca atggcagaca ataacgaagg cgccgacgga 660 gtgggtaatg cctcaggaaa ttggcattgc gattccacat ggctggggcga cagagtcatc 720 accaccagca cccgaacctg ggccctcccc acctacaaca accacctcta caagcaaatc 780 tccaacagcc aatcgggagg aagcaccaac gacaacacct acttcggcta cagcaccccc 840 tggggtatt ttgactttaa cagattccac tgccacttct caccacgtga ctggcagcga 900 ctcatcaaca acaactgggg attccggccc aagagactca acttcaagct cttcaacatc 960 caggtcaagg aggtcacgac gaatgatggc accacgacca tcgccaataa ccttaccagc 1020 acggttcagg tctttacgga ctcggaatac cagctcccgt acgtcctcgg ctctgcgcac 1080 cagggctgcc tgcctccgtt cccggcggac gtcttcatga ttcctcagta cgggtacctg 1140 actctgaaca atggcagtca ggccgtgggc cgttcctcct tctactgcct ggagtacttt 1200 ccttctcaaa tgctgagaac gggcaacaac tttgagttca gctacacgtt tgaggacgtg 1260 ccttttcaca gcagctacgc gcacagccaa agcctggacc ggctgatgaa ccccctcatc 1320 gaccagtacc tgtactacct gtctcggact cagaccacgg gaggtaccgc aggaaatcgg 1380 acgttgcaat tttctcaggc cgggcctagt agcatggcga atcaggccaa aaactggcta 1440 cccgggccct gctaccggca gcaacgcgtc tccaagacaa cgaatcaaaa taacaacagc 1500 aactttgcct ggaccggtgc caccaagtat catctgaatg gcagagactc tctggtaaat 1560 cccggtgtcg ctatggcaac ccacaaggac gacgaagacc gattttttcc gtccagcgga 1620 gtcttaatat ttgggaaaca gggagctgga aatgacaacg tggaccttga caacgttatg 1680 ataaccagtg aggaagaaat taaaaccacc aacccagtgg ccacagaaga gtacggcgtg 1740 gtggccacta acctgcaatc ggcaaacacc gctcctcaaa cagggaccgt caacagtcaa 1800 ggagccttac ctggcatggt ctggcagaac cgggacgtgt acctgcaggg tcctatctgg 1860 gccaagattc ctcacacgga cggaaacttt catccctcgc cgctgatggg aggctttgga 1920 ctgaaacacc cgcctcctca gatcctgatt aagaatacac ctgttcccgc gaatcctcca 1980 actaccttca gtccagctaa gtttgcgtcg ttcatcacgc agtacagcac cggacaggtc 2040 agcgtggaaa ttgaatggga gctgcagaaa gaaaacagca aacgctggaa cccagagatt 2100 caatacactt ccaactacaa caaatctaca aatgtggact ttgctgttga cacagaaggc 2160 gtttattctg agcctcgccc catcggcacc cgttacctca cccgtaatct g 2211 <210> 5 <211> 738 <212> PRT <213> Adeno-associated virus <220> <221> Variants <222> (158)..(158) <223> / Replace="Ser" <220> <221> Variants <222> (169)..(169) <223> / Replace="Arg" <220> <221> Variants <222> (564)..(564) <223> / Replace="Asn" <220> <221> misc_feature <222> (1)..(738) <223> / note="There is no preference for variant residues given in the sequence relative to those in the annotation at the variant position" <400> 5 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Arg Glu Pro Asp Ser Ser Thr Gly Ile 145 150 155 160 Gly Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln 165 170 175 Thr Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro 180 185 190 Pro Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ala Gly Gly 195 200 205 Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn 210 215 220 Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val 225 230 235 240 Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His 245 250 255 Leu Tyr Lys Gln Ile Ser Asn Gly Thr Ser Gly Gly Ser Thr Asn Asp 260 265 270 Asn Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn 275 280 285 Arg Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn 290 295 300 Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn 305 310 315 320 Ile Gln Val Lys Glu Val Thr Thr Asn Glu Gly Thr Lys Thr Ile Ala 325 330 335 Asn Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln 340 345 350 Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe 355 360 365 Pro Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn 370 375 380 Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr 385 390 395 400 Phe Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr 405 410 415 Thr Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser 420 425 430 Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu 435 440 445 Ser Arg Thr Gln Thr Thr Gly Gly Thr Ala Gly Thr Gln Thr Leu Gln 450 455 460 Phe Ser Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp 465 470 475 480 Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Thr Thr Thr Asn 485 490 495 Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His 500 505 510 Leu Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr 515 520 525 His Lys Asp Asp Glu Asp Arg Phe Phe Pro Ser Ser Gly Val Leu Ile 530 535 540 Phe Gly Lys Gln Gly Ala Gly Asn Asp Asn Val Asp Tyr Ser Asn Val 545 550 555 560 Met Ile Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr 565 570 575 Glu Glu Tyr Gly Val Val Ala Thr Asn Leu Gln Ser Ala Asn Thr Ala 580 585 590 Pro Gln Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val 595 600 605 Trp Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile 610 615 620 Pro His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe 625 630 635 640 Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val 645 650 655 Pro Ala Asp Pro Pro Thr Thr Phe Asn Gln Ala Lys Leu Asn Ser Phe 660 665 670 Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu 675 680 685 Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr 690 695 700 Ser Asn Tyr Tyr Lys Ser Thr Asn Val Asp Phe Ala Val Asn Thr Glu 705 710 715 720 Gly Val Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg 725 730 735 Asn Leu <210> 6 <211> 2214 <212> DNA <213> Adeno-associated virus <220> <221> Mutations <222> (472)..(474) <223> / replace="agc" <220> <221> Mutations <222> (505)..(507) <223> / replace="aga" <220> <221> Mutations <222> (1690)..(1692) <223> / replace="aac" <220> <221> misc_feature <222> (1)..(2214) <223> / note="There is no preference for variant nucleotides given in the sequence relative to those in the variant position annotation" <400> 6 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acctgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata atcacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 ggaaagaaga gaccggtaga gcagtcacca cagcgtgagc ccgactcctc cacgggcatc 480 ggcaagaaag gccagcagcc cgccaaaaag agactcaatt tcggtcagac tggcgactca 540 gagtcagtcc ccgaccctca acctctcgga gaacctccag cagcgccctc tggtgtggga 600 tctaatacaa tggctgcagg cggtggcgca ccaatggcag acaataacga aggtgccgac 660 ggagtgggta attcctcggg aaattggcat tgcgattcca catggctggg cgacagagtc 720 atcaccacca gcacccgaac ctgggccctg cccacctaca acaaccacct ctacaagcaa 780 atctccaacg ggacctcggg aggcagcacc aacgacaaca cctactttgg ctacagcacc 840 ccctggggt attttgactt taacagattc cactgccact tctcaccacg tgactggcag 900 cgactcatca acaacaactg gggattccgg cccaagagac tcaacttcaa gctcttcaac 960 atccaggtca aagaggtcac gacgaatgaa ggcaccaaga ccatcgccaa taacctcacc 1020 agcaccgtcc aggtgtttac ggactcggaa taccagctgc cgtacgtcct cggctctgcc 1080 caccagggct gcctgcctcc gttcccggcg gacgtcttca tgattcctca gtacggctac 1140 ctgactctca acaacggtag tcaggccgtg ggacgttcct ccttctactg cctggagtac 1200 ttcccctctc agatgctgag aacgggcaac aactttcaat tcagctacac tttcgaggac 1260 gtgcctttcc acagcagcta cgcgcacagc cagagtttgg acaggctgat gaatcctctc 1320 atcgaccagt acctgtacta cctgtcaaga acccagacta cgggaggcac agcgggaacc 1380 cagacgttgc agttttctca ggccgggcct agcagcatgg cgaatcaggc caaaaactgg 1440 ctgcctggac cctgctacag acagcagcgc gtctccacga caacgaatca aaacaacaac 1500 agcaactttg cctggactgg tgccaccaag tatcatctga acggcagaga ctctctggtg 1560 aatccgggcg tcgccatggc aacccacaag gacgacgagg accgcttctt cccatccagc 1620 ggcgtcctca tatttggcaa gcagggagct ggaaatgaca acgtggacta tagcaacgtg 1680 atgataacca gcgaggaaga aatcaagacc accaaccccg tggccacaga agagtatggc 1740 gtggtggcta ctaacctaca gtcggcaaac accgctcctc aaacggggac cgtcaacagc 1800 cagggagcct tacctggcat ggtctggcag aaccgggacg tgtacctgca gggtcctatt 1860 tgggccaaga ttcctcacac agatggcaac tttcacccgt ctcctttaat gggcggcttt 1920 ggacttaaac atccgcctcc tcagatcctc atcaaaaaca ctcctgttcc tgcggatcct 1980 ccaacaacgt tcaaccaggc caagctgaat tctttcatca cgcagtacag caccggacaa 2040 gtcagcgtgg agatcgagtg ggagctgcag aaggagaaca gcaagcgctg gaacccagag 2100 attcagtata cttccaacta ctacaaatct acaaatgtgg actttgctgt taatactgag 2160 ggtgtttact ctgagcctcg ccccattggc actcgttacc tcacccgtaa tctg 2214 <210> 7 <211> 738 <212> PRT <213> Adeno-associated virus <220> <221> Variants <222> (158)..(158) <223> / Replace="Ser" <220> <221> Variants <222> (169)..(169) <223> / replace="Lys" <220> <221> Variants <222> (315)..(315) <223> / Replace="Ser" <220> <221> Variants <222> (413)..(413) <223> / Replace="Glu" <220> <221> Variants <222> (472)..(472) <223> / Replace="Thr" or "Ser" <220> <221> Variants <222> (534)..(534) <223> / Replace="Glu" <220> <221> Variants <222> (542)..(542) <223> / Replace="Val" <220> <221> Variants <222> (595)..(595) <223> / Replace="Val" <220> <221> misc_feature <222> (1)..(738) <223> / note="There is no preference for variant residues given in the sequence relative to those in the annotation at the variant position" <400> 7 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Arg Glu Pro Asp Ser Ser Thr Gly Ile 145 150 155 160 Gly Lys Lys Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln 165 170 175 Thr Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro 180 185 190 Pro Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ala Gly Gly 195 200 205 Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser 210 215 220 Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val 225 230 235 240 Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His 245 250 255 Leu Tyr Lys Gln Ile Ser Asn Gly Thr Ser Gly Gly Ser Thr Asn Asp 260 265 270 Asn Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn 275 280 285 Arg Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn 290 295 300 Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn 305 310 315 320 Ile Gln Val Lys Glu Val Thr Gln Asn Glu Gly Thr Lys Thr Ile Ala 325 330 335 Asn Asn Leu Thr Ser Thr Ile Gln Val Phe Thr Asp Ser Glu Tyr Gln 340 345 350 Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe 355 360 365 Pro Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn 370 375 380 Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr 385 390 395 400 Phe Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr 405 410 415 Thr Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser 420 425 430 Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu 435 440 445 Ser Arg Thr Gln Thr Thr Gly Gly Thr Ala Gly Thr Gln Thr Leu Gln 450 455 460 Phe Ser Gln Ala Gly Pro Ser Asn Met Ala Asn Gln Ala Lys Asn Trp 465 470 475 480 Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Thr Thr Thr Ser 485 490 495 Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His 500 505 510 Leu Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr 515 520 525 His Lys Asp Asp Glu Asp Arg Phe Phe Pro Ser Ser Gly Ile Leu Ile 530 535 540 Phe Gly Lys Gln Gly Ala Gly Lys Asp Asn Val Asp Tyr Ser Asn Val 545 550 555 560 Met Leu Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr 565 570 575 Glu Glu Tyr Gly Val Val Ala Asp Asn Leu Gln Gln Gln Asn Thr Ala 580 585 590 Pro Gln Ile Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val 595 600 605 Trp Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile 610 615 620 Pro His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe 625 630 635 640 Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val 645 650 655 Pro Ala Asp Pro Pro Thr Thr Phe Asn Gln Ala Lys Leu Asn Ser Phe 660 665 670 Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu 675 680 685 Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr 690 695 700 Ser Asn Tyr Tyr Lys Ser Thr Asn Val Asp Phe Ala Val Asn Thr Glu 705 710 715 720 Gly Val Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg 725 730 735 Asn Leu <210> 8 <211> 2214 <212> DNA <213> Adeno-associated virus <220> <221> Mutations <222> (472)..(474) <223> / replace="agc" <220> <221> Mutations <222> (505)..(507) <223> / replace="aag" <220> <221> Mutations <222> (943)..(945) <223> / replace="agc" <220> <221> Mutations <222> (1237)..(1239) <223> / replace="gaa" <220> <221> Mutations <222> (1414)..(1416) <223> / Replace="aac" or "agc" <220> <221> Mutations <222> (1600)..(1602) <223> / replace="gag" <220> <221> Mutations <222> (1624)..(1626) <223> / replace="gtc" <220> <221> Mutations <222> (1783)..(1785) <223> / replace="gta" <220> <221> misc_feature <222> (1)..(2214) <223> / note="There is no preference for variant nucleotides given in the sequence relative to those in the variant position annotation" <400> 8 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acctgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata atcacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 ggaaagaaga gaccggtaga gcagtcacca cagcgtgagc ccgactcctc cacgggcatc 480 ggcaagaaag gccagcagcc cgccagaaag agactcaatt tcggtcagac tggcgactca 540 gagtcagtcc ccgaccctca acctctcgga gaacctccag cagcgccctc tggtgtggga 600 tctaatacaa tggctgcagg cggtggcgca ccaatggcag acaataacga aggtgccgac 660 ggagtgggta gttcctcggg aaattggcat tgcgattcca catggctggg cgacagagtc 720 atcaccacca gcacccgaac ctgggccctg cccacctaca acaaccacct ctacaagcaa 780 atctccaacg ggacctcggg aggcagcacc aacgacaaca cctactttgg ctacagcacc 840 ccctggggt attttgactt taacagattc cactgccact tctcaccacg tgactggcag 900 cgactcatca acaacaactg gggattccgg cccaagagac tcaacttcaa gctcttcaac 960 atccaggtca aagaggtcac gcagaatgaa ggcaccaaga ccatcgccaa taacctcacc 1020 agcaccatcc aggtgtttac ggactcggaa taccagctgc cgtacgtcct cggctctgcc 1080 caccagggct gcctgcctcc gttcccggcg gacgtcttca tgattcctca gtacggctac 1140 ctgactctca acaacggtag tcaggccgtg ggacgttcct ccttctactg cctggagtac 1200 ttcccctctc agatgctgag aacgggcaac aactttcaat tcagctacac tttcgaggac 1260 gtgcctttcc acagcagcta cgcgcacagc cagagtttgg acaggctgat gaatcctctc 1320 atcgaccagt acctgtacta cctgtcaaga acccagacta cgggaggcac agcgggaacc 1380 cagacgttgc agttttctca ggccgggcct agcaacatgg cgaatcaggc caaaaactgg 1440 ctgcctggac cctgctacag acagcagcgc gtctccacga caacgtcgca aaacaacaac 1500 agcaactttg cctggactgg tgccaccaag tatcatctga acggcagaga ctctctggtg 1560 aatccgggcg tcgccatggc aacccacaag gacgacgagg accgcttctt cccatccagc 1620 ggcatcctca tatttggcaa gcagggagct ggaaaagaca acgtggacta tagcaacgtg 1680 atgctaacca gcgaggaaga aatcaagacc accaaccccg tggccacaga agagtatggc 1740 gtggtggctg ataacctaca gcagcaaaac accgctcctc aaatagggac cgtcaacagc 1800 cagggagcct tacctggcat ggtctggcag aaccgggacg tgtacctgca gggtcctatt 1860 tgggccaaga ttcctcacac agatggcaac tttcacccgt ctcctttaat gggcggcttt 1920 ggacttaaac atccgcctcc tcagatcctc atcaaaaaca ctcctgttcc tgcggatcct 1980 ccaacaacgt tcaaccaggc caagctgaat tctttcatca cgcagtacag caccggacaa 2040 gtcagcgtgg agatcgagtg ggagctgcag aaggagaaca gcaagcgctg gaacccagag 2100 attcagtata cttccaacta ctacaaatct acaaatgtgg actttgctgt taatactgag 2160 ggtgtttact ctgagcctcg ccccattggc actcgttacc tcacccgtaa tctg 2214 <210> 9 <211> 738 <212> PRT <213> Adeno-associated virus <220> <221> Variants <222> (169)..(169) <223> / replace="Lys" <220> <221> Variants <222> (315)..(315) <223> / Replace="Ser" <220> <221> Variants <222> (534)..(534) <223> / Replace="Glu" <220> <221> Variants <222> (542)..(542) <223> / Replace="Val" <220> <221> misc_feature <222> (1)..(738) <223> / note="There is no preference for variant residues given in the sequence relative to those in the annotation at the variant position" <400> 9 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Pro Ser Pro Gln Arg Ser Pro Asp Ser Ser Thr Gly Ile 145 150 155 160 Gly Lys Lys Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln 165 170 175 Thr Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro 180 185 190 Pro Ala Ala Pro Ser Gly Val Gly Ser Gly Thr Met Ala Ala Gly Gly 195 200 205 Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser 210 215 220 Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val 225 230 235 240 Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His 245 250 255 Leu Tyr Lys Gln Ile Ser Asn Gly Thr Ser Gly Gly Ser Thr Asn Asp 260 265 270 Asn Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn 275 280 285 Arg Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn 290 295 300 Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn 305 310 315 320 Ile Gln Val Lys Glu Val Thr Gln Asn Glu Gly Thr Lys Thr Ile Ala 325 330 335 Asn Asn Leu Thr Ser Thr Ile Gln Val Phe Thr Asp Ser Glu Tyr Gln 340 345 350 Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe 355 360 365 Pro Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn 370 375 380 Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr 385 390 395 400 Phe Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr 405 410 415 Thr Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser 420 425 430 Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu 435 440 445 Ser Arg Thr Gln Ser Thr Gly Gly Thr Ala Gly Thr Gln Gln Leu Leu 450 455 460 Phe Ser Gln Ala Gly Pro Ser Asn Met Ser Ala Gln Ala Lys Asn Trp 465 470 475 480 Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Thr Thr Leu Ser 485 490 495 Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His 500 505 510 Leu Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr 515 520 525 His Lys Asp Asp Glu Asp Arg Phe Phe Pro Ser Ser Gly Ile Leu Met 530 535 540 Phe Gly Lys Gln Gly Ala Gly Lys Asp Asn Val Asp Tyr Ser Asn Val 545 550 555 560 Met Leu Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr 565 570 575 Glu Gln Tyr Gly Val Val Ala Asp Asn Leu Gln Gln Gln Asn Thr Ala 580 585 590 Pro Ile Val Gly Ala Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val 595 600 605 Trp Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile 610 615 620 Pro His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe 625 630 635 640 Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val 645 650 655 Pro Ala Asp Pro Pro Thr Thr Phe Asn Gln Ala Lys Leu Asn Ser Phe 660 665 670 Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu 675 680 685 Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr 690 695 700 Ser Asn Tyr Tyr Lys Ser Thr Asn Val Asp Phe Ala Val Asn Thr Glu 705 710 715 720 Gly Val Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg 725 730 735 Asn Leu <210> 10 <211> 2214 <212> DNA <213> Adeno-associated virus <220> <221> Mutations <222> (505)..(507) <223> / replace="aaa" <220> <221> Mutations <222> (943)..(945) <223> / replace="agc" <220> <221> Mutations <222> (1600)..(1602) <223> / replace="gag" <220> <221> Mutations <222> (1624)..(1626) <223> / replace="gtc" <220> <221> misc_feature <222> (1)..(2214) <223> / note="There is no preference for variant nucleotides given in the sequence relative to those in the variant position annotation" <400> 10 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acctgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata atcacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 ggaaagaaga gaccggtaga gccgtcacca cagcgttccc ccgactcctc cacgggcatc 480 ggcaagaaag gccagcagcc cgccagaaag agactcaatt tcggtcagac tggcgactca 540 gagtcagtcc ccgaccctca acctatcgga gaacctccag cagcgccctc tggtgtggga 600 tctggtacaa tgggctcagg cggtggcgca ccaatggcag acataga aggtgccgac 660 ggagtgggta gttcctcggg aaattggcat tgcgattcca catggctggg cgacagagtc 720 atcaccacca gcacccgaac ctgggccctg cccacctaca acaaccacct ctacaagcaa 780 atctccaacg ggacctcggg aggcagcacc aacgacaaca cctactttgg ctacagcacc 840 ccctggggt attttgactt taacagattc cactgccact tctcaccacg tgactggcag 900 cgactcatca acaacaactg gggattccgg cccaagagac tcaacttcaa gctcttcaac 960 atccaggtca aagaggtcac gcagaatgaa ggcaccaaga ccatcgccaa taacctcacc 1020 agcaccatcc aggtgtttac ggactcggaa taccagctgc cgtacgtcct cggctctgcc 1080 caccagggct gcctgcctcc gttcccggcg gacgtcttca tgattcctca gtacggctac 1140 ctgactctca acaacggtag tcaggccgtg ggacgttcct ccttctactg cctggagtac 1200 ttcccctctc agatgctgag aacgggcaac aactttgagt tcagctacac tttcgaggac 1260 gtgcctttcc acagcagcta cgcgcacagc cagagtttgg acaggctgat gaatcctctc 1320 atcgaccagt acctgtacta cctgtcaaga acccagtcta cgggaggcac agcgggaacc 1380 cagcagttgc tgttttctca ggccgggcct agcaacatgt cggctcaggc caaaaactgg 1440 ctgcctggac cctgctacag acagcagcgc gtctccacga cactgtcgca aaacaacaac 1500 agcaactttg cctggactgg tgccaccaag tatcatctga acggcagaga ctctctggtg 1560 aatccgggcg tcgccatggc aacccacaag gacgacgagg accgcttctt cccatccagc 1620 ggcatcctca tgtttggcaa gcagggagct ggaaaagaca acgtggacta tagcaacgtg 1680 atgctaacca gcgaggaaga aatcaagacc accaaccccg tggccacaga acagtatggc 1740 gtggtggctg ataacctaca gcagcaaaac accgctccta ttgtgggggc cgtcaacagc 1800 cagggagcct tacctggcat ggtctggcag aaccgggacg tgtacctgca gggtcctatt 1860 tgggccaaga ttcctcacac agatggcaac tttcacccgt ctcctttaat gggcggcttt 1920 ggacttaaac atccgcctcc tcagatcctc atcaaaaaca ctcctgttcc tgcggatcct 1980 ccaacaacgt tcaaccaggc caagctgaat tctttcatca cgcagtacag caccggacaa 2040 gtcagcgtgg agatcgagtg ggagctgcag aaggagaaca gcaagcgctg gaacccagag 2100 attcagtata cttccaacta ctacaaatct acaaatgtgg actttgctgt taatactgag 2160 ggtgtttact ctgagcctcg ccccattggc actcgttacc tcacccgtaa tctg 2214 <210> 11 <211> 738 <212> PRT <213> Adeno-associated virus <220> <221> Variants <222> (471)..(471) <223> / Replace="Asn" <220> <221> misc_feature <222> (1)..(738) <223> / note="There is no preference for variant residues given in the sequence relative to those in the annotation at the variant position" <400> 11 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Pro Ser Pro Gln Arg Ser Pro Asp Ser Ser Thr Gly Ile 145 150 155 160 Gly Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln 165 170 175 Thr Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro 180 185 190 Pro Ala Gly Pro Ser Gly Leu Gly Ser Gly Thr Met Ala Ala Gly Gly 195 200 205 Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser 210 215 220 Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val 225 230 235 240 Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His 245 250 255 Leu Tyr Lys Gln Ile Ser Asn Gly Thr Ser Gly Gly Ser Thr Asn Asp 260 265 270 Asn Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn 275 280 285 Arg Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn 290 295 300 Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn 305 310 315 320 Ile Gln Val Lys Glu Val Thr Gln Asn Glu Gly Thr Lys Thr Ile Ala 325 330 335 Asn Asn Leu Thr Ser Thr Ile Gln Val Phe Thr Asp Ser Glu Tyr Gln 340 345 350 Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe 355 360 365 Pro Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn 370 375 380 Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr 385 390 395 400 Phe Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr 405 410 415 Thr Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser 420 425 430 Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu 435 440 445 Ser Arg Thr Gln Ser Thr Gly Gly Thr Ala Gly Thr Gln Gln Leu Leu 450 455 460 Phe Ser Gln Ala Gly Pro Ser Asn Met Ser Ala Gln Ala Lys Asn Trp 465 470 475 480 Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Thr Thr Leu Ser 485 490 495 Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His 500 505 510 Leu Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr 515 520 525 His Lys Asp Asp Glu Glu Arg Phe Phe Pro Ser Ser Gly Val Leu Met 530 535 540 Phe Gly Lys Gln Gly Ala Gly Lys Asp Asn Val Asp Tyr Ser Ser Val 545 550 555 560 Met Leu Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr 565 570 575 Glu Gln Tyr Gly Val Val Ala Asp Asn Leu Gln Gln Gln Asn Thr Ala 580 585 590 Pro Ile Val Gly Ala Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val 595 600 605 Trp Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile 610 615 620 Pro His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe 625 630 635 640 Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val 645 650 655 Pro Ala Asp Pro Pro Thr Thr Phe Ser Gln Ala Lys Leu Ala Ser Phe 660 665 670 Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu 675 680 685 Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr 690 695 700 Ser Asn Tyr Tyr Lys Ser Thr Asn Val Asp Phe Ala Val Asn Thr Glu 705 710 715 720 Gly Thr Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg 725 730 735 Asn Leu <210> 12 <211> 2214 <212> DNA <213> Adeno-associated virus <220> <221> Mutations <222> (1411)..(1413) <223> / replace="aat" <220> <221> misc_feature <222> (1)..(2214) <223> / note="There is no preference for variant nucleotides given in the sequence relative to those in the annotation at the variant position" <400> 12 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acttgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata accacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 ggaaagaaga gaccggtaga gccatcaccc cagcgttctc cagactcctc tacgggcatc 480 ggcaagaaag gccagcagcc cgcgaaaaag agactcaact ttgggcagac tggcgactca 540 gagtcagtgc ccgaccctca accaatcgga gaaccccccg caggcccctc tggtctggga 600 tctggtacaa tggctgcagg cggtggcgct ccaatggcag acaataacga aggcgccgac 660 ggagtgggta gttcctcagg aaattggcat tgcgattcca catggctggg cgacagagtc 720 atcaccacca gcacccgaac ctgggccctc cccacctaca acaaccacct ctacaagcaa 780 atctccaacg ggacttcggg aggaagcacc aacgacaaca cctacttcgg ctacagcacc 840 ccctgggggt attttgactt taacagattc cactgccact tctcaccacg tgactggcag cgactcatca acaacaactg gggattccgg cccaagagac tcaacttcaa gctcttcaac atccaggtca aggaggtcac gcagaatgaa ggcaccaaga ccatcgccaa taaccttacc agcacgattc aggtctttac ggactcggaa taccagctcc cgtacgtcct cggctctgcg caccagggct gcctgcctcc gttcccggcg gacgtcttca tgattcctca gtacgggtac 1140 ctgactctga acaatggcag tcaggccgtg ggccgttcct ccttctactg cctggagtac tttccttctc aaatgctgag aacgggcaac aactttgagt tcagctacac gtttgaggac gtgccttttc acagcagcta cgcgcacagc caaagcctgg accggctgat gaaccccctc 1320 atcgaccagt acctgtacta cctgtctcgg actcagtcca cgggaggtac cgcaggaact cagcagttgc tattttctca ggccggggcct agtaacatgt cggctcaggc caaaaactgg ctacccgggc cctgctaccg gcagcaacgc gtctccacga cactgtcgca aaataacaac agcaactttg cctggaccgg tgccaccaag tatcatctga atggcagaga ctctctggta aatcccggtg tcgctatggc aacccacaag gacgacgaag agcgattttt tccgtccagc 1620 ggagtcttaa tgtttgggaa acagggagct ggaaaagaca acgtggacta tagcagcgtt 1680 atgctaacca gtgaggaaga aattaaaacc accaacccag tggccacaga acagtacggc 1740 gtggtggccg ataacctgca acagcaaaac accgctccta ttgtaggggc cgtcaacagt 1800 caaggagcct tacctggcat ggtctggcag aaccgggacg tgtacctgca gggtcctatc 1860 tgggccaaga ttcctcacac ggacggaaac tttcatccct cgccgctgat gggaggcttt 1920 ggactgaaac acccgcctcc tcagatcctg attaagaata cacctgttcc cgcggatcct 1980[[ID=1,3]] ccaactacct tcagtcaagc taagctggcg tcgttcatca cgcagtacag caccggacag 2040 gtcagcgtgg aaattgaatg ggagctgcag aaagaaaaca gcaaacgctg gaacccagag 2100 attcaataca cttccaacta ctacaaatct acaaatgtgg actttgctgt taacacagaa 2160 ggcacttatt ctgagcctcg ccccatcggc acccgttacc tcacccgtaa tctg 2214 <210> 13 <211> 737 <212> PRT <213> Adeno - associated virus <220> <221> Variants <222> (148)..(148) <223> / Replace="Gln" <220> <221> Variants <222> (169)..(169) <223> / Replace="Arg" <220> <221> Variants <222> (314)..(314) <223> / Replace="Asn" <220> <221> Variants <222> (466)..(466) <223> / Replace="His" <220> <221> Variants <222> (563)..(563) <223> / Replace="Ser" <220> <221> Variants <222> (580)..(580) <223> / Replace="Ile" <220> <221> Variants <222> (588)..(588) <223> / Replace="Ser" <220> <221> misc_feature <222> (1)..(737) <223> / note="There is no preference for variant residues given in the sequence relative to those in the annotation at the variant position" <400> 13 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Pro Ser Pro Gln Arg Ser Pro Asp Ser Ser Thr Gly Ile 145 150 155 160 Gly Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln 165 170 175 Thr Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro 180 185 190 Pro Ala Ala Pro Ser Gly Val Gly Ser Gly Thr Met Ala Ala Gly Gly 195 200 205 Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn 210 215 220 Ala Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val 225 230 235 240 Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His 245 250 255 Leu Tyr Lys Gln Ile Ser Ser Gln Ser Ala Gly Ser Thr Asn Asp Asn 260 265 270 Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg 275 280 285 Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn 290 295 300 Asn Trp Gly Phe Arg Pro Lys Lys Leu Arg Phe Lys Leu Phe Asn Ile 305 310 315 320 Gln Val Lys Glu Val Thr Thr Asn Asp Gly Val Thr Thr Ile Ala Asn 325 330 335 Asn Leu Thr Ser Thr Val Gln Val Phe Ser Asp Ser Glu Tyr Gln Leu 340 345 350 Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro 355 360 365 Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn 370 375 380 Gly Ser Gln Ser Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe 385 390 395 400 Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Thr 405 410 415 Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu 420 425 430 Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ala 435 440 445 Arg Thr Gln Ser Thr Thr Gly Gly Thr Ala Gly Asn Arg Glu Leu Gln 450 455 460 Phe Tyr Gln Ala Gly Pro Ser Thr Met Ala Glu Gln Ala Lys Asn Trp 465 470 475 480 Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Leu Asp 485 490 495 Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His 500 505 510 Leu Asn Gly Arg Asn Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr 515 520 525 His Lys Asp Asp Glu Asp Arg Phe Phe Pro Ser Ser Gly Val Leu Ile 530 535 540 Phe Gly Lys Thr Gly Ala Ala Asn Lys Thr Thr Leu Glu Asn Val Leu 545 550 555 560 Met Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu 565 570 575 Glu Tyr Gly Val Val Ser Ser Asn Leu Gln Ser Ala Asn Thr Ala Pro 580 585 590 Gln Thr Gln Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp 595 600 605 Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro 610 615 620 His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly 625 630 635 640 Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro 645 650 655 Ala Asn Pro Pro Glu Val Phe Thr Pro Ala Lys Phe Ala Ser Phe Ile 660 665 670 Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu 675 680 685 Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser 690 695 700 Asn Tyr Asp Lys Ser Thr Asn Val Asp Phe Ala Val Asp Ser Glu Gly 705 710 715 720 Val Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn 725 730 735 Leu <210> 14 <211> 2211 <212> DNA <213> Adeno-associated virus <220> <221> Mutations <222> (442)..(444) <223> / replace="cag" <220> <221> Mutations <222> (505)..(507) <223> / replace="aga" <220> <221> Mutations <222> (940)..(942) <223> / replace="aac" <220> <221> Mutations <222> (1396)..(1398) <223> / replace="cac" <220> <221> Mutations <222> (1687)..(1689) <223> / replace="agt" <220> <221> Mutations <222> (1738)..(1740) <223> / replace="ata" <220> <221> Mutations <222> (1762)..(1764) <223> / replace="tct" <220> <221> misc_feature <222> (1)..(2211) <223> / note="There is no preference for variant nucleotides given in the sequence relative to those in the variant position annotation" <400> 14 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acctgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata accacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtcattt gggggcaacc tcgggcgagc agtcttccag 360 gccaagaagc gggttctcga acctcggt ctggttgagg aaggcgctaa gacggctcct 420 ggaaagaaga gaccggtaga gccgtcacct cagcgttccc ccgactcctc cacgggcatc 480 ggcaagaaag gccagcagcc cgccaaaaag agactcaatt tcggtcagac tggcgactca 540 gagtcagtcc ccgaccctca acctctcgga gaacctccag cagcgccctc tggtgtggga 600 tctggtacaa tggctgcagg cggtggcgca ccaatggcag acaataacga aggtgccgac 660 ggagtgggta atgcctcagg aaattggcat tgcgattcca catggctggg cgacagagtc 720 attaccacca gcacccgaac ctgggccctg cccacctaca acaaccacct ctacaagcaa 780 atctccagtc aaagtgcagg tagtaccaac gacaacacct acttcggcta cagcaccccc 840 tgggggtatt ttgactttaa cagattccac tgccacttct caccacgtga ctggcagcga 900 ctcatcaaca acaactgggg attccggccc aagaagctgc ggttcaagct cttcaacatc 960 caggtcaagg aggtcacgac gaatgacggc gttacgacca tcgctaataa ccttaccagc 1020 acggttcagg tattctcgga ctcggaatac cagctgccgt acgtcctcgg ctctgcgcac 1080 cagggctgcc tgcctccgtt cccggcggac gtcttcatga ttcctcagta cggctacctg 1140 actctcaaca atggcagtca gtctgtggga cgttcctcct tctactgcct ggagtacttc 1200 ccctctcaga tgctgagaac gggcaacaac tttgagttca gctacacctt cgaggacgtg 1260 cctttccaca gcagctacgc acacagccag agcctggacc ggctgatgaa tcccctcatc 1320 gaccagtact tgtactacct ggccagaaca cagagtacca caggaggcac agctggcaat 1380 cgggaactgc agttttacca ggccgggcct tcaactatgg ccgaacaagc caagaattgg 1440 ttacctggac cttgctaccg gcaacaaaga gtctccaaaa cgctggatca aaacaacaac 1500 agcaactttg cttggactgg tgccaccaaa tatcacctga acggcagaaa ctcgttggtt 1560 aatcccggcg tcgccatggc aactcacaag gacgacgagg accgcttttt cccatccagc 1620 ggagtcctga tttttggaaa aactggagca gctaacaaaa ctacattgga aaatgtgtta 1680 atgacaaatg aagaagaaat taaaactact aatcctgtag ccacggaaga atacggggta 1740 gtcagcagca acttacaatc ggctaatact gcaccccaga cacaaactgt caacagccag 1800 ggagccttac ctggcatggt ctggcagaac cgggacgtgt acctgcaggg tcccatctgg 1860 gccaagattc ctcacacgga tggcaacttt cacccgtctc ctttgatggg cggctttgga 1920 cttaaacatc cgcctcctca gatcctgatc aagaacactc ccgttcccgc taatcctccg 1980 gaggtgttta ctcctgccaa gtttgcttcg ttcatcacac agtacagcac cggacaagtc 2040 agcgtggaaa tcgagtggga gctgcagaag gaaaacagca agcgctggaa cccggagatt 2100 cagtacacct ccaactatga taagtcgact aatgtggact ttgccgttga cagcgagggt 2160 gtttactctg agcctcgccc tattggcact cgttacctca cccgtaatct g 2211 <210> 15 <211> 735 <212> PRT <213> Adeno-associated virus <220> <221> Variants <222> (162)..(162) <223> / Replace="Thr" <220> <221> Variants <222> (168)..(168) <223> / Replace="Arg" <220> <221> Variants <222> (224)..(224) <223> / Replace="Ser" <220> <221> Variants <222> (310)..(310) <223> / replace="Lys" <220> <221> Variants <222> (410)..(410) <223> / Replace="Gln" <220> <221> Variants <222> (446)..(446) <223> / Replace="Asn" <220> <221> Variants <222> (461)..(461) <223> / replace="Leu" <220> <221> Variants <222> (471)..(471) <223> / Replace="Ser" <220> <221> Variants <222> (708)..(708) <223> / Replace="Thr" <220> <221> misc_feature <222> (1)..(735) <223> / note="There is no preference for variant residues given in the sequence relative to those in the variant position annotation" <400> 15 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Ser Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala Ser Asn Asp Asn His Tyr 260 265 270 Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe His 275 280 285 Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn Trp 290 295 300 Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln Val 305 310 315 320 Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn Leu 325 330 335 Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr 340 345 350 Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala Asp 355 360 365 Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser 370 375 380 Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro Ser 385 390 395 400 Gln Met Leu Arg Thr Gly Asn Asn Phe Thr Phe Ser Tyr Thr Phe Glu 405 410 415 Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg 420 425 430 Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg Thr 435 440 445 Gln Thr Thr Ser Gly Thr Ala Gln Asn Arg Glu Leu Gln Phe Ser Gln 450 455 460 Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu Pro Gly 465 470 475 480 Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ala Asn Asp Asn Asn 485 490 495 Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn Gly 500 505 510 Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Ser His Lys Asp 515 520 525 Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Leu Ile Phe Gly Lys 530 535 540 Gln Gly Ala Gly Ala Ser Asn Val Asp Leu Asp Asn Val Met Ile Thr 545 550 555 560 Asp Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr 565 570 575 Gly Thr Val Ala Thr Asn Leu Gln Ser Ser Asn Thr Ala Pro Ala Thr 580 585 590 Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln Asp 595 600 605 Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His Thr 610 615 620 Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu Lys 625 630 635 640 His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala Asn 645 650 655 Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr Gln 660 665 670 Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln Lys 675 680 685 Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn Tyr 690 695 700 Asn Lys Ser Ala Asn Val Asp Phe Thr Val Asp Thr Asn Gly Val Tyr 705 710 715 720 Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 16 <211> 2205 <212> DNA <213> Adeno-associated virus <220> <221> Mutations <222> (484)..(486) <223> / replace="aca" <220> <221> Mutations <222> (502)..(504) <223> / replace="aga" <220> <221> Mutations <222> (670)..(672) <223> / replace="tcc" <220> <221> Mutations <222> (928)..(930) <223> / replace="aaa" <220> <221> Mutations <222> (1228)..(1230) <223> / replace="cag" <220> <221> Mutations <222> (1336)..(1338) <223> / replace="aac" <220> <221> Mutations <222> (1381)..(1383) <223> / replace="ctg" <220> <221> Mutations <222> (1411)..(1413) <223> / replace="tct" <220> <221> Mutations <222> (2122)..(2124) <223> / replace="acc" <220> <221> misc_feature <222> (1)..(2205) <223> / note="There is no preference for variant nucleotides given in the sequence relative to those in the variant position annotation" <400> 16 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acttgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggatgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata accacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 gccaaaaga gggttctcga acctcttggt ctggttgagg aaggtgctaa gacggctcct 420 ggaaagaaac gtccggtaga gcagtcgcca caagagccag actcctcctc gggcattggc 480 aagtcaggcc agcagcccgc taaaaagaga ctcaattttg gtcagactgg cgactcagag 540 tcagtccccg acccacaacc tctcggagaa cctccagcag ccccctctgg tgtgggatct 600 aatacaatgg cttcaggcgg tggcgcacca atggcagaca ataacgaagg cgccgacgga 660 gtgggtaatg cctcaggaaa ttggcattgc gattccacat ggctggggcga cagagtcatc 720 accaccagca cccgaacatg ggccttgccc acctataaca accacctcta caagcaaatc 780 tccagtcaat caggggccag caaccaac cactacttcg gctacaggcac cccctgggg 840 tattttgatt tcaacagatt ccactgccat ttctcaccac gtgactggca gcgactcatc 900 aacaacaatt ggggattccg gcccaagaga ctcaacttca agctcttcaa catccaagtc 960 aaggaggtca cgacgaatga tggcaccacg accatcgcta ataaccttac cagcacggtt 1020 caagtcttca cggactcgga gtaccagttg ccgtacgtcc tcggctctgc gcaccagggc 1080 tgcctccctc cgttcccggc ggacgtgttc atgattccgc agtacggcta cctaacgctc 1140 aacaatggca gccaggcagt gggacggtca tccttttact gcctggaata tttcccatcg 1200 cagatgctga gaacgggcaa taactttacc ttcagctaca ccttcgagga cgtgcctttc 1260 cacagcagct acgcgcacag ccagagcctg gaccggctga tgaatcctct catcgaccag 1320 tacctgtatt acctgagcag aactcagact acgtccggaa ctgcccaaaa cagggagttg 1380 cagtttagcc aggcgggtcc atctagcatg gctaatcagg ccaaaaactg gctacctgga 1440 ccctgttacc ggcagcagcg cgtttctaaa acagcaaatg acaacaacaa cagcaacttt 1500 gcctggactg gtgctacaaa atatcacctt aatgggcgtg attctttagt caaccctggc 1560 cctgctatgg cctcacacaa agacgacgaa gacaagttct ttcccatgag cggtgtcttg 1620 atttttggaa agcagggcgc cggagcttca aacgttgatt tggacaatgt catgatcaca 1680 gacgaagagg aaatcaaaac cactaacccc gtggccaccg aacaatatgg gactgtggca 1740 accaatctcc agagcagcaa cacagcccct gcgaccggaa ctgtgaattc tcagggagcc 1800 ttacctggaa tggtgtggca agacagagac gtatacctgc agggtcctat ttgggccaaa 1860 attcctcaca cggatggaca ctttcacccg tctcctctca tgggcggctt tggacttaag 1920 cacccgcctc ctcagatcct catcaaaaac acgcctgttc ctgcgaatcc tccgacaacg 1980 ttttcgcctg caaagtttgc ttcattcatc acccagtatt ccacaggaca agtgagcgtg 2040 gagattgaat gggagctgca gaaagaaaac agcaaacgct ggaatcccga aatacagtat 2100 acatctaact ataataaatc tgccaacgtt gatttcactg tggacaccaa tggagtttat 2160 agtgagcctc gccccattgg cacccgttac ctcacccgta acctg 2205 <210> 17 <211> 735 <212> PRT <213> Adeno-associated virus <220> <221> Variants <222> (42)..(42) <223> / Replace="Ser" <220> <221> Variants <222> (168)..(168) <223> / replace="Lys" <220> <221> Variants <222> (310)..(310) <223> / Replace="Arg" <220> <221> Variants <222> (410)..(410) <223> / Replace="Gln" <220> <221> Variants <222> (446)..(446) <223> / Replace="Arg" <220> <221> Variants <222> (461)..(461) <223> / replace="Leu" <220> <221> Variants <222> (471)..(471) <223> / Replace="Ser" <220> <221> Variants <222> (475)..(475) <223> / Replace="Arg" <220> <221> Variants <222> (504)..(504) <223> / Replace="Ala" <220> <221> Variants <222> (539)..(539) <223> / Replace="Asn" <220> <221> misc_feature <222> (1)..(735) <223> / note="There is no preference for variant residues given in the sequence relative to those in the variant position annotation" <400> 17 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Gln Pro 20 25 30 Lys Ala Asn Gln Gln His Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Glu Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Ser Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ser 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala Ser Asn Asp Asn His Tyr 260 265 270 Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe His 275 280 285 Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn Trp 290 295 300 Gly Phe Arg Pro Lys Lys Leu Asn Phe Lys Leu Phe Asn Ile Gln Val 305 310 315 320 Lys Glu Val Thr Gln Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn Leu 325 330 335 Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr 340 345 350 Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala Asp 355 360 365 Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser 370 375 380 Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro Ser 385 390 395 400 Gln Met Leu Arg Thr Gly Asn Asn Phe Thr Phe Ser Tyr Thr Phe Glu 405 410 415 Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg 420 425 430 Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg Thr 435 440 445 Gln Thr Thr Ser Gly Thr Thr Gln Gln Ser Arg Leu Gln Phe Ser Gln 450 455 460 Ala Gly Pro Ser Ser Met Ala Gln Gln Ala Lys Asn Trp Leu Pro Gly 465 470 475 480 Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ala Asn Asp Asn Asn 485 490 495 Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn Gly 500 505 510 Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Ser His Lys Asp 515 520 525 Asp Glu Glu Lys Phe Phe Pro Met His Gly Val Leu Ile Phe Gly Lys 530 535 540 Gln Gly Thr Gly Ala Ser Asn Val Asp Leu Asp Asn Val Met Ile Thr 545 550 555 560 Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr 565 570 575 Gly Thr Val Ala Thr Asn Leu Gln Ser Ser Asn Thr Ala Pro Ala Thr 580 585 590 Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln Asp 595 600 605 Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His Thr 610 615 620 Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu Lys 625 630 635 640 His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala Asn 645 650 655 Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr Gln 660 665 670 Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln Lys 675 680 685 Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn Tyr 690 695 700 Asn Lys Ser Val Asn Val Asp Phe Thr Val Asp Thr Asn Gly Val Tyr 705 710 715 720 Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 18 <211> 2205 <212> DNA <213> Adeno-associated virus <220> <221> Mutations <222> (124)..(126) <223> / replace="agt" <220> <221> Mutations <222> (502)..(504) <223> / replace="aaa" <220> <221> Mutations <222> (928)..(930) <223> / replace="aga" <220> <221> Mutations <222> (1228)..(1230) <223> / replace="cag" <220> <221> Mutations <222> (1336)..(1338) <223> / replace="aga" <220> <221> Mutations <222> (1381)..(1383) <223> / replace="ctc" <220> <221> Mutations <222> (1411)..(1413) <223> / replace="tct" <220> <221> Mutations <222> (1423)..(1425) <223> / replace="aga" <220> <221> Mutations <222> (1510)..(1512) <223> / replace="gcg" <220> <221> Mutations <222> (1615)..(1617) <223> / replace="gac" <220> <221> misc_feature <222> (1)..(2205) <223> / note="There is no preference for variant nucleotides given in the sequence relative to those in the variant position annotation" <400> 18 atggctgctg acggttatct tccagattgg ctcgaggaca acctttctga aggcattcgt 60 gagtggtggg atctgaaacc tggagcccct caacccaaag cgaaccaaca acaccaggac 120 gacggtcggg gtcttgtgct tccgggttac aaatacctcg gaccctttaa cggactcgac 180 aaaggagagc cggtcaacga ggcggacgcg gcagccctcg aacacgacaa agcttacgac 240 cagcagctca aggccggtga caacccgtac ctcaagtaca accacgccga cgccgagttt 300 caggagcgtc ttcaagaaga tacgtctttt gggggcaacc ttggcagagc agtcttccag 360 gccaaaaaga gggtccttga gcctcttggt ctggttgagg aagcagctaa aacggctcct 420 ggaaagaaga ggcctgtaga acagtctcct caggaaccgg actcatcatc tggtattggc 480 aaatcgggcc aacagcctgc cagaaaaaga ctaaatttcg gtcagactgg agactcagag 540 tcagtcccag accctcaacc tctcggagaa ccaccagcag ccccctcagg tgtgggatct 600 aatacaatgg cttcaggcgg tggcgcacca atggcagaca ataacgaggg tgccgatgga 660 gtgggtaatt cctcaggaaa ttggcattgc gattccacat ggctgggcga cagagtcatc 720 accaccagca ccagaacctg ggccctgccc acttacaaca accatctcta caagcaaatc 780 tccagccaat caggagcttc aaacgacaac cactactttg gctacagcac cccttggggg 840 tattttgact ttaacagatt ccactgccac ttctcaccac gtgactggca gcgactcatt 900 aacaacaact ggggattccg gcccaagaaa ctcaacttca agctcttcaa catccaagtt 960 aaagaggtca cgcagaacga tggcacgacg actattgcca ataaccttac cagcacggtt 1020 caagtgttta cggactcgga gtatcagctc ccgtacgtgc tcgggtcggc gcaccaaggc 1080 tgtctcccgc cgtttccagc ggacgtctttc atgatccctc agtatggata cctcaccctg 1140 aacaacggaa gtcaagcggt gggacgctca tccttttact gcctggagta cttcccttcg 1200 cagatgctaa ggactggaaa taacttcaca ttcagctata ccttcgagga tgtacctttt 1260 cacagcagct acgctcacag ccagagtttg gatcgcttga tgaatcctct tattgatcag 1320 tatctgtact acctgagcag aacgcaaaca acctctggaa caacccaaca atcacggctg 1380 caatttagcc aggctgggcc ttcgtctatg gctcagcagg ccaaaaattg gctacctggg 1440 ccctgctacc ggcaacagag agtttcaaag actgctaacg acaacaacaa cagtaacttt 1500 gctggacag gggccaccaa atatcatctc aatggccgcg actcgctggt gaatccagga 1560 ccagctatgg ccagtcacaa ggacgatgaa gaaaaatttt tccctatgca cggcgttcta 1620 atatttggca aacaagggac aggggcaagt aacgtagatt tagataatgt aatgattacg 1680 gatgaagaag agattcgtac caccaatcct gtggcaacag agcagtatgg aactgtggca 1740 actaacttgc agagctcaaa tacagctccc gcgactggaa ctgtcaatag tcagggggcc 1800 ttacctggca tggtgtggca agatcgtgac gtgtaccttc aaggacctat ctgggcaaag 1860 attcctcaca cggatggaca ctttcatcct tctcctctga tgggaggctt tggactgaaa 1920 catccgcctc ctcaaatctt gatcaaaaat actccggtac cggcaaatcc tccgacgact 1980 ttcagcccgg ccaagtttgc ttcatttatc actcagtact ccactggaca ggtcagcgtg 2040 gaaattgagt gggagctaca gaaagaaaac agcaaacgtt ggaatccaga gattcagtac 2100 acttccaact acaacaagtc tgttaatgtg gactttactg tagacactaa tggtgtttat 2160 agtgaacctc gccctattgg aacccggtat ctcacacgaa acttg 2205 `<210> 19 <211> 736 <212> PRT <213> Adeno - associated virus <400> 19 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Lys Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ala Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Gly Ser Thr Asn Asp Asn Thr 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Thr Phe 405 410 415 Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg 435 440 445 Thr Gln Thr Thr Ser Gly Thr Ala Gly Asn Arg Thr Leu Gln Phe Ser 450 455 460 Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ala Asn Gln Asn 485 490 495 Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Thr His Lys 515 520 525 Asp Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Leu Ile Phe Gly 530 535 540 Lys Gln Gly Ala Gly Asn Ser Asn Val Asp Leu Asp Asn Val Met Ile 545 550 555 560 Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Gln 565 570 575 Tyr Gly Thr Val Ala Thr Asn Leu Gln Ser Ala Asn Thr Ala Pro Ala 580 585 590 Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Thr Asn Val Asp Phe Ala Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 20 <211> 736 <212> PRT <213> Adeno-associated virus <400> 20 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala Ser Thr Asn Asp Asn Thr 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Thr Phe 405 410 415 Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg 435 440 445 Thr Gln Thr Thr Ser Gly Thr Ala Gly Asn Arg Glu Leu Gln Phe Ser 450 455 460 Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Thr Asn Gln Asn 485 490 495 Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Thr His Lys 515 520 525 Asp Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Leu Ile Phe Gly 530 535 540 Lys Gln Gly Ala Gly Asn Ser Asn Val Asp Leu Asp Asn Val Met Ile 545 550 555 560 Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Glu 565 570 575 Tyr Gly Thr Val Ala Thr Asn Leu Gln Ser Ala Asn Thr Ala Pro Ala 580 585 590 Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Thr Asn Val Asp Phe Ala Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 21 <211> 736 <212> PRT [[ID=**21**]]<213> Adeno-associated virus <400> 21 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Lys Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ala Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Gly Ser Thr Asn Asp Asn Thr 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Thr Phe 405 410 415 Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg 435 440 445 Thr Gln Thr Thr Ser Gly Thr Ala Gly Asn Arg Glu Leu Gln Phe Ser 450 455 460 Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Thr Asn Gln Asn 485 490 495 Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Thr His Lys 515 520 525 Asp Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Leu Ile Phe Gly 530 535 540 Lys Gln Gly Ala Gly Asn Ser Asn Val Asp Leu Asp Asn Val Met Ile 545 550 555 560 Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Glu 565 570 575 Tyr Gly Thr Val Ala Thr Asn Leu Gln Ser Ser Asn Thr Ala Pro Ala 580 585 590 Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Glu Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Thr Asn Val Asp Phe Ala Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 22 <211> 736 <212> PRT <213> Adeno-associated virus <400> 22 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30[[ID=2,4]] Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 。 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 注:原文中“50 55 60 ”后的句号是中文句号,可能是原文格式问题,翻译时保留。若有特殊要求,请根据实际情况调整。 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Lys Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Gly Ser Thr Asn Asp Asn Thr 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Lys Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Thr Phe 405 410 415 Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg 435 440 445 Thr Gln Thr Thr Ser Gly Thr Ala Gly Asn Arg Glu Leu Gln Phe Ser 450 455 460 Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Thr Asn Gln Asn 485 490 495 Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Thr His Lys 515 520 525 Asp Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Leu Ile Phe Gly 530 535 540 Lys Gln Gly Ala Gly Asn Ser Asn Val Asp Leu Asp Asn Val Met Ile 545 550 555 560 Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Glu 565 570 575 Tyr Gly Thr Val Ala Thr Asn Leu Gln Ser Ala Asn Thr Ala Pro Ala 580 585 590 Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Thr Asn Val Asp Phe Ala Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 23 <211> 736 <212> PRT <213> Adeno-associated virus <400> 23 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 [[ID=(32]]Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly It should be noted that there seems to be a small formatting issue in the original text where the tag for ID 32 has an extra closing parenthesis. I've translated it as presented while keeping this in mind. 145 150 155 160 Lys Lys Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ala Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Gly Ser Thr Asn Asp Asn Thr 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Lys Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Thr Phe 405 410 415 Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg 435 440 445 Thr Gln Thr Thr Ser Gly Thr Ala Gly Asn Arg Thr Leu Gln Phe Ser 450 455 460 Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Thr Asn Gln Asn 485 490 495 Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Thr His Lys 515 520 525 Asp Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Leu Ile Phe Gly 530 535 540 Lys Gln Gly Ala Gly Asn Ser Asn Val Asp Leu Asp Asn Val Met Ile 545 550 555 560 Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Glu 565 570 575 Tyr Gly Thr Val Ala Thr Asn Leu Gln Ser Ala Asn Thr Ala Pro Ala 580 585 590 Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Thr Asn Val Asp Phe Ala Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 24 <--!> 736 <212> PRT <--!> Adeno-associated virus <400> 24 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Note: There seems to be an issue with the original text where the tag <211> has "736" which might be an incorrect format. I've translated it as <--!> 736 and for <213> "腺伴随病毒" I've translated as <--!> Adeno-associated virus following the pattern of the other tags in the text. If this is not what is expected, please clarify the correct format or meaning of these tags.Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ala Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Gly Ser Thr Asn Asp Asn Thr 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Lys Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Thr Phe 405 410 415 Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg 435 440 445 Thr Gln Thr Thr Ser Gly Thr Ala Gly Asn Arg Thr Leu Gln Phe Ser 450 455 460 Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ala Asn Gln Asn 485 490 495 Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Thr His Lys 515 520 525 Asp Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Leu Ile Phe Gly 530 535 540 Lys Gln Gly Ala Gly Asn Ser Asn Val Asp Leu Asp Asn Val Met Ile 545 550 555 560 Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Gln 565 570 575 Tyr Gly Thr Val Ala Thr Asn Leu Gln Ser Ser Asn Thr Ala Pro Ala 580 585 590 Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Thr Asn Val Asp Phe Ala Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 25 <211> 736 <212> PRT <213> Adeno-associated virus <400> 25 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Gly Ser Thr Asn Asp Asn Thr 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Lys Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Thr Phe 405 410 415 Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg 435 440 445 Thr Gln Thr Thr Ser Gly Thr Ala Gly Asn Arg Thr Leu Gln Phe Ser 450 455 460 Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ala Asn Gln Asn 485 490 495 Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Thr His Lys 515 520 525 Asp Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Leu Ile Phe Gly 530 535 540 Lys Gln Gly Ala Gly Asn Ser Asn Val Asp Leu Asp Asn Val Met Ile 545 550 555 560 Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Glu 565 570 575 Tyr Gly Thr Val Ala Thr Asn Leu Gln Ser Ser Asn Thr Ala Pro Ala 580 585 590 Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Thr Asn Val Asp Phe Ala Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 26 <211> 7 "36" <212> PRT <213> Adeno - associated virus <400> 26 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala Note: There seems to be an issue with the "7 "36"" in the translation of <211> tag. It's not clear if the space and the double quotes are part of the original content or an error. If it's an error, it should probably be "736". 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Asn Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Gly Ser Thr Asn Asp Asn Thr 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Lys Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Thr Phe 405 410 415 Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg 435 440 445 Thr Gln Thr Thr Ser Gly Thr Ala Gly Asn Arg Glu Leu Gln Phe Ser 450 455 460 Gln Ala Gly Pro Ser Ser Met Ala Asn Gln Ala Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Thr Asn Gln Asn 485 490 495 Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Thr His Lys 515 520 525 Asp Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Leu Ile Phe Gly 530 535 540 Lys Gln Gly Ala Gly Asn Ser Asn Val Asp Leu Asp Asn Val Met Ile 545 550 555 560 Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Gln 565 570 575 Tyr Gly Thr Val Ala Thr Asn Leu Gln Ser Ala Asn Thr Ala Pro Ala 580 585 590 Thr Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Thr Asn Val Asp Phe Ala Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 27 <211> 738 <212> PRT <213> virus <400> 27 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Gln Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Pro Ser Pro Gln Arg Ser Pro Asp Ser Ser Thr Gly Ile 145 150 155 160 Gly Lys Lys Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln 165 170 175 Thr Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro 180 185 190 Pro Ala Ala Pro Ser Gly Val Gly Pro Asn Thr Met Ala Ala Gly Gly 195 200 205 Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser 210 215 220 Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val 225 230 235 240 Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His 245 250 255 Leu Tyr Lys Gln Ile Ser Asn Gly Thr Ser Gly Gly Ala Thr Asn Asp 260 265 270 Asn Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn 275 280 285 Arg Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn 290 295 300 Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Ser Phe Lys Leu Phe Asn 305 310 315 320 Ile Gln Val Lys Glu Val Thr Gln Asn Glu Gly Thr Lys Thr Ile Ala 325 330 335 Asn Asn Leu Thr Ser Thr Ile Gln Val Phe Thr Asp Ser Glu Tyr Gln 340 345 350 Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe 355 360 365 Pro Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn 370 375 380 Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr 385 390 395 400 Phe Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Thr Tyr 405 410 415 Thr Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser 420 425 430 Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu 435 440 445 Ser Arg Thr Gln Thr Thr Gly Gly Thr Ala Asn Thr Gln Thr Leu Gly 450 455 460 Phe Ser Gln Gly Gly Pro Asn Thr Met Ala Asn Gln Ala Lys Asn Trp 465 470 475 480 Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Thr Thr Thr Gly 485 490 495 Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Ala Gly Thr Lys Tyr His 500 505 510 Leu Asn Gly Arg Asn Ser Leu Ala Asn Pro Gly Ile Ala Met Ala Thr 515 520 525 His Lys Asp Asp Glu Glu Arg Phe Phe Pro Ser Asn Gly Ile Leu Ile 530 535 540 Phe Gly Lys Gln Asn Ala Ala Arg Asp Asn Ala Asp Tyr Ser Asp Val 545 550 555 560 Met Leu Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr 565 570 575 Glu Glu Tyr Gly Ile Val Ala Asp Asn Leu Gln Gln Gln Asn Thr Ala 580 585 590 Pro Gln Ile Gly Thr Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val 595 600 605 Trp Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile 610 615 620 Pro His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe 625 630 635 640 Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val 645 650 655 Pro Ala Asp Pro Pro Thr Thr Phe Asn Gln Ser Lys Leu Asn Ser Phe 660 665 670 Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu 675 680 685 Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr 690 695 700 Ser Asn Tyr Tyr Lys Ser Thr Ser Val Asp Phe Ala Val Asn Thr Glu 705 710 715 720 Gly Val Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg 725 730 735 Asn Leu <210> 28 <211> 736 <212> PRT <213> Adeno-associated virus <400> 28 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Ala Pro Gln Pro 20 25 30 Lys Ala Asn Gln Gln His Gln Asp Asn Ala Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Leu Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ala Gly Ile Gly 145 150 155 160 Lys Ser Gly Ala Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Thr Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro Pro 180 185 190 Ala Ala Pro Ser Gly Val Gly Ser Leu Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Val Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser Ser 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Asn Ser Thr Ser Gly Gly Ser Ser Asn Asp Asn 260 265 270 Ala Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg 275 280 285 Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn 290 295 300 Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile 305 310 315 320 Gln Val Lys Glu Val Thr Asp Asn Asn Gly Val Lys Thr Ile Ala Asn 325 330 335 Asn Leu Thr Ser Thr Val Gln Val Phe Thr Asp Ser Asp Tyr Gln Leu 340 345 350 Pro Tyr Val Leu Gly Ser Ala His Glu Gly Cys Leu Pro Pro Phe Pro 355 360 365 Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asp 370 375 380 Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe 385 390 395 400 Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Glu 405 410 415 Phe Glu Asn Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu 420 425 430 Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser 435 440 445 Lys Thr Ile Asn Gly Ser Gly Gln Asn Gln Gln Thr Leu Lys Phe Ser 450 455 460 Val Ala Gly Pro Ser Asn Met Ala Val Gln Gly Arg Asn Tyr Ile Pro 465 470 475 480 Gly Pro Ser Tyr Arg Gln Gln Arg Val Ser Thr Thr Val Thr Gln Asn 485 490 495 Asn Asn Ser Glu Phe Ala Trp Pro Gly Ala Ser Ser Trp Ala Leu Asn 500 505 510 Gly Arg Asn Ser Leu Met Asn Pro Gly Pro Ala Met Ala Ser His Lys 515 520 525 Glu Gly Glu Asp Arg Phe Phe Pro Leu Ser Gly Ser Leu Ile Phe Gly 530 535 540 Lys Gln Gly Thr Gly Arg Asp Asn Val Asp Ala Asp Lys Val Met Ile 545 550 555 560 Thr Asn Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Ser 565 570 575 Tyr Gly Gln Val Ala Thr Asn His Gln Ser Ala Gln Ala Gln Ala Gln 580 585 590 Thr Gly Trp Val Gln Asn Gln Gly Ile Leu Pro Gly Met Val Trp Gln 595 600 605 Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Met 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asp Pro Pro Thr Ala Phe Asn Lys Asp Lys Leu Asn Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Tyr Lys Ser Asn Asn Val Glu Phe Ala Val Asn Thr Glu Gly Val 705 7l0 l15 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 29 <211> 736 <212> PRT <213> Adeno - associated virus [[ID=~39]]<400> 29 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Phe Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Thr Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Thr Pro Ala Ala Val Gly Pro Thr Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Ala Ser Thr Gly Ala Ser Asn Asp Asn His 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Val Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Ser Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Thr Phe Ser Tyr Thr Phe 405 410 415 Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Asn Arg 435 440 445 Thr Gln Asn Gln Ser Gly Ser Ala Gln Asn Lys Asp Leu Leu Phe Ser 450 455 460 Arg Gly Ser Pro Ala Gly Met Ser Val Gln Pro Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Lys Thr Asp Asn 485 490 495 Asn Asn Ser Asn Phe Thr Trp Thr Gly Ala Ser Lys Tyr Asn Leu Asn 500 505 510 Gly Arg Glu Ser Ile Ile Asn Pro Gly Thr Ala Met Ala Ser His Lys 515 520 525 Asp Asp Lys Asp Lys Phe Phe Pro Met Ser Gly Val Met Ile Phe Gly 530 535 540 Lys Glu Ser Ala Gly Ala Ser Asn Thr Ala Leu Asp Asn Val Met Ile 545 550 555 560 Thr Asp Glu Glu Glu Ile Lys Ala Thr Asn Pro Val Ala Thr Glu Arg 565 570 575 Phe Gly Thr Val Ala Val Asn Leu Gln Ser Ser Ser Thr Asp Pro Ala 580 585 590 Thr Gly Asp Val His Val Met Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Ala Glu Phe Ser Ala Thr Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Val Gln Tyr Thr Ser Asn 690 695 700 Tyr Ala Lys Ser Ala Asn Val Asp Phe Thr Val Asp Asn Asn Gly Leu 705 710 715 720 Tyr Thr Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Pro Leu 725 730 735 <210> 30 <211> 736 <212> PRT <213> Adeno-associated virus <400> 30 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 2S 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Ile Gly 145 150 155 160 Lys Thr Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Thr Pro Ala Ala Val Gly Pro Thr Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ala 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Ala Ser Thr Gly Ala Ser Asn Asp Asn His 260 265 270 Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe 275 280 285 His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn 290 295 300 Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln 305 310 315 320 Val Lys Glu Val Thr Thr Asn Asp Gly Val Thr Thr Ile Ala Asn Asn 325 330 335 Leu Thr Ser Thr Val Gln Val Phe Ser Asp Ser Glu Tyr Gln Leu Pro 340 345 350 Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala 355 360 365 Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly 370 375 380 Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro 385 390 395 400 Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Thr Phe Ser Tyr Thr Phe 405 410 415 Glu Glu Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp 420 425 430 Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Asn Arg 435 440 445 Thr Gln Asn Gln Ser Gly Ser Ala Gln Asn Lys Asp Leu Leu Phe Ser 450 455 460 Arg Gly Ser Pro Ala Gly Met Ser Val Gln Pro Lys Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Lys Thr Asp Asn 485 490 495 Asn Asn Ser Asn Phe Thr Trp Thr Gly Ala Ser Lys Tyr Asn Leu Asn 500 505 510 Gly Arg Glu Ser Ile Ile Asn Pro Gly Thr Ala Met Ala Ser His Lys 515 520 525 Asp Asp Glu Asp Lys Phe Phe Pro Met Ser Gly Val Met Ile Phe Gly 530 535 540 Lys Glu Ser Ala Gly Ala Ser Asn Thr Ala Leu Asp Asn Val Met Ile 545 550 555 560 Thr Asp Glu Glu Glu Ile Lys Ala Thr Asn Pro Val Ala Thr Glu Arg 565 570 575 Phe Gly Thr Val Ala Val Asn Phe Gln Ser Ser Ser Thr Asp Pro Ala 580 585 590 Thr Gly Asp Val His Ala Met Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys Asn Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Ala Glu Phe Ser Ala Thr Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Val Gln Tyr Thr Ser Asn 690 695 700 Tyr Ala Lys Ser Ala Asn Val Asp Phe Thr Val Asp Asn Asn Gly Leu 705 710 715 720 Tyr Thr Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Pro Leu 725 730 735 <210> 31 <211> 735 <212> PRT <213> Adeno-associated virus <400> 31 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Thr Leu Ser 1 5 10 15 Glu Gly Ile Arg Gln Trp Trp Lys Leu Lys Pro Gly Pro Pro Pro Pro 20 25 30 Lys Pro Ala Glu Arg His Lys Asp Asp Ser Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Glu Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Arg Gln Leu Asp Ser Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Lys Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Pro Val Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu His Ser Pro Val Glu Pro Asp Ser Ser Ser Gly Thr Gly 145 150 155 160 Lys Ala Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ala Asp Ser Val Pro Asp Pro Gln Pro Leu Gly Gln Pro Pro 180 185 190 Ala Ala Pro Ser Gly Leu Gly Thr Asn Thr Met Ala Thr Gly Ser Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ser 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Thr Trp Met Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala Ser Asn Asp Asn His Tyr 260 265 270 Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe His 275 280 285 Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn Trp 290 295 300 Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn Ile Gln Val 305 310 315 320 Lys Glu Val Thr Gln Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn Leu 325 330 335 Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr 340 345 350 Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala Asp 355 360 365 Val Phe Met Val Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser 370 375 380 Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro Ser 385 390 395 400 Gln Met Leu Arg Thr Gly Asn Asn Phe Thr Phe Ser Tyr Thr Phe Glu 405 410 415 Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg 420 425 430 Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ser Arg Thr 435 440 445 Asn Thr Pro Ser Gly Thr Thr Thr Gln Ser Arg Leu Gln Phe Ser Gln 450 455 460 Ala Gly Ala Ser Asp Ile Arg Asp Gln Ser Arg Asn Trp Leu Pro Gly 465 470 475 480 Pro Cys Tyr Arg Gln Gln Arg Val Ser Lys Thr Ser Ala Asp Asn Asn 485 490 495 Asn Ser Glu Tyr Ser Trp Thr Gly Ala Thr Lys Tyr His Leu Asn Gly 500 505 510 Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Ser His Lys Asp 515 520 525 Asp Glu Glu Lys Phe Phe Pro Gln Ser Gly Val Leu Ile Phe Gly Lys 530 535 540 Gln Gly Ser Glu Lys Thr Asn Val Asp Ile Glu Lys Val Met Ile Thr 545 550 555 560 Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr 565 570 575 Gly Ser Val Ser Thr Asn Leu Gln Arg Gly Asn Arg Gln Ala Ala Thr 580 585 590 Ala Asp Val Asn Thr Gln Gly Val Leu Pro Gly Met Val Trp Gln Asp 595 600 605 Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His Thr 610 615 620 Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu Lys 625 630 635 640 His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro Ala Asn 645 650 655 Pro Ser Thr Thr Phe Ser Ala Ala Lys Phe Ala Ser Phe Ile Thr Gln 660 665 670 Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln Lys 675 680 685 Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn Tyr 690 695 700 Asn Lys Ser Val Asn Val Asp Phe Thr Val Asp Thr Asn Gly Val Tyr 705 710 715 720 Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 32 <211> 736 <212> PRT <213> Adeno-associated virus <400> 32 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Val Pro Gln Pro 20 25 30 Lys Ala Asn Gln Gln His Gln Asp Asn Arg Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Glu Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Ile Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Gly 130 135 140 Ala Val Asp Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Val Gly 145 150 155 160 Lys Ser Gly Lys Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Thr Ser Leu Gly Ser Asn Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ser 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala Ser Asn Asp Asn His Tyr 260 265 270 Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe His 275 280 285 Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn Trp 290 295 300 Gly Phe Arg Pro Lys Lys Leu Ser Phe Lys Leu Phe Asn Ile Gln Val 305 310 315 320 Arg Gly Val Thr Gln Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn Leu 325 330 335 Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr 340 345 350 Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala Asp 355 360 365 Val Phe Met Val Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser 370 375 380 Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro Ser 385 390 395 400 Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Thr Phe Glu 405 410 415 Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg 420 425 430 Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Asn Arg Thr 435 440 445 Gln Gly Thr Thr Ser Gly Thr Thr Asn Gln Ser Arg Leu Leu Phe Ser 450 455 460 Gln Ala Gly Pro Gln Ser Met Ser Leu Gln Ala Arg Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Leu Ser Lys Thr Ala Asn Asp Asn 485 490 495 Asn Asn Ser Asn Phe Pro Trp Thr Ala Ala Ser Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Ser His Lys 515 520 525 Asp Asp Glu Glu Lys Phe Phe Pro Met His Gly Asn Leu Ile Phe Gly 530 535 540 Lys Glu Gly Thr Thr Ala Ser Asn Ala Glu Leu Asp Asn Val Met Ile 545 550 555 560 Thr Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu Gln 565 570 575 Tyr Gly Thr Val Ala Asn Asn Leu Gln Ser Ser Asn Thr Ala Pro Thr 580 585 590 Thr Gly Thr Val Asn His Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Met Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Val Asn Val Asp Phe Thr Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735 <210> 33 <211> 736 <212> PRT <213> Adeno-associated virus <400> 33 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Ala Leu Lys Pro Gly Val Pro Gln Pro 20 25 30 Lys Ala Asn Gln Gln His Gln Asp Asn Arg Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Gly Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Glu Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Lys Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Ile Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Ala Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Asp Gln Ser Pro Gln Glu Pro Asp Ser Ser Ser Gly Val Gly 145 150 155 160 Lys Ser Gly Lys Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln Thr 165 170 175 Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro Pro 180 185 190 Ala Ala Pro Thr Ser Leu Gly Ser Asn Thr Met Ala Ser Gly Gly Gly 195 200 205 Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn Ser 210 215 220 Ser Gly Asn Trp His Cys Asp Ser Gln Trp Leu Gly Asp Arg Val Ile 225 230 235 240 Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His Leu 245 250 255 Tyr Lys Gln Ile Ser Ser Gln Ser Gly Ala Ser Asn Asp Asn His Tyr 260 265 270 Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg Phe His 275 280 285 Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn Asn Trp 290 295 300 Gly Phe Arg Pro Lys Lys Leu Ser Phe Lys Leu Phe Asn Ile Gln Val 305 310 315 320 Lys Glu Val Thr Gln Asn Asp Gly Thr Thr Thr Ile Ala Asn Asn Leu 325 330 335 Thr Ser Thr Val Gln Val Phe Thr Asp Ser Glu Tyr Gln Leu Pro Tyr 340 345 350 Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro Ala Asp 355 360 365 Val Phe Met Val Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn Gly Ser 370 375 380 Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe Pro Ser 385 390 395 400 Gln Met Leu Arg Thr Gly Asn Asn Phe Gln Phe Ser Tyr Thr Phe Glu 405 410 415 Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu Asp Arg 420 425 430 Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Asn Arg Thr 435 440 445 Gln Gly Thr Thr Ser Gly Thr Thr Asn Gln Ser Arg Leu Leu Phe Ser 450 455 460 Gln Ala Gly Pro Gln Ser Met Ser Leu Gln Ala Arg Asn Trp Leu Pro 465 470 475 480 Gly Pro Cys Tyr Arg Gln Gln Arg Leu Ser Lys Thr Ala Asn Asp Asn 485 490 495 Asn Asn Ser Asn Phe Pro Trp Thr Ala Ala Ser Lys Tyr His Leu Asn 500 505 510 Gly Arg Asp Ser Leu Val Asn Pro Gly Pro Ala Met Ala Ser His Lys 515 520 525 Asp Asp Glu Glu Lys Phe Phe Pro Met His Gly Asn Leu Ile Phe Gly 530 535 540 Lys Glu Gly Thr Thr Ala Ser Asn Ala Glu Leu Asp Asn Val Met Ile 545 550 555 560 Thr Asp Glu Glu Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu Gln 565 570 575 Tyr Gly Thr Val Ala Asn Asn Leu Gln Ser Ser Asn Thr Ala Pro Thr 580 585 590 Thr Arg Thr Val Asn Asp Gln Gly Ala Leu Pro Gly Met Val Trp Gln 595 600 605 Asp Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro His 610 615 620 Thr Asp Gly His Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly Leu 625 630 635 640 Lys His Pro Pro Pro Gln Ile Met Ile Lys Asn Thr Pro Val Pro Ala 645 650 655 Asn Pro Pro Thr Thr Phe Ser Pro Ala Lys Phe Ala Ser Phe Ile Thr 660 665 670 Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu Gln 675 680 685 Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser Asn 690 695 700 Tyr Asn Lys Ser Val Asn Val Asp Phe Thr Val Asp Thr Asn Gly Val 705 710 715 720 Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn Leu 725 730 735<000408۴><210> 34 <211> 737 <212> PRT <2۱۳> Adeno-associated virus <400> 34 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asn Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Ala Lys Lys Arg 130 135 140 Pro Val Glu Pro Ser Pro Gln Arg Ser Pro Asp Ser Ser Thr Gly Ile 145 150 155 160 Gly Lys Lys Gly Gln Gln Pro Ala Arg Lys Arg Leu Asn Phe Gly Gln 165 170 175 Thr Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Leu Gly Glu Pro 180 185 190 Pro Ala Ala Pro Ser Ser Val Gly Ser Gly Thr Val Ala Ala Gly Gly 195 200 205 Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Asn 210 215 220 Ala Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val 225 230 235 240 Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His 245 250 255 Leu Tyr Lys Gln Ile Ser Ser Glu Thr Ala Gly Ser Thr Asn Asp Asn 260 265 270 Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn Arg 275 280 285 Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn Asn 290 295 300 Asn Trp Gly Phe Arg Pro Lys Lys Leu Arg Phe Lys Leu Phe Asn Ile 305 310 315 320 Gln Val Lys Glu Val Thr Thr Asn Asp Gly Val Thr Thr Ile Ala Asn 325 330 335 Asn Leu Thr Ser Thr Ile Gln Val Phe Ser Asp Ser Glu Tyr Gln Leu 340 345 350 Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe Pro 355 360 365 Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn Asn 370 375 380 Gly Ser Gln Ser Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr Phe 385 390 395 400 Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr Ser 405 410 415 Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser Leu 420 425 430 Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu Ala 435 440 445 Arg Thr Gln Ser Asn Pro Gly Gly Thr Ala Gly Asn Arg Glu Leu Gln 450 455 460 Phe Tyr Gln Gly Gly Pro Ser Thr Met Ala Glu Gln Ala Lys Asn Trp 465 470 475 480 Leu Pro Gly Pro Cys Phe Arg Gln Gln Arg Val Ser Lys Thr Leu Asp 485 490 495 Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His 500 505 510 Leu Asn Gly Arg Asn Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr 515 520 525 His Lys Asp Asp Glu Asp Arg Phe Phe Pro Ser Ser Gly Val Leu Ile 530 535 540 Phe Gly Lys Thr Gly Ala Thr Asn Lys Thr Thr Leu Glu Asn Val Leu 545 550 555 560 Met Thr Asn Glu Glu Glu Ile Arg Pro Thr Asn Pro Val Ala Thr Glu 565 570 575 Glu Tyr Gly Ile Val Ser Ser Asn Leu Gln Ala Ala Asn Thr Ala Ala 580 585 590 Gln Thr Gln Val Val Asn Asn Gln Gly Ala Leu Pro Gly Met Val Trp 595 600 605 Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile Pro 610 615 620 His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe Gly 625 630 635 640 Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val Pro 645 650 655 Ala Asn Pro Pro Glu Val Phe Thr Pro Ala Lys Phe Ala Ser Phe Ile 660 665 670 Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu Leu 675 680 685 Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr Ser 690 695 700 Asn Phe Glu Lys Gln Thr Gly Val Asp Phe Ala Val Asp Ser Gln Gly 705 710 715 720 Val Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg Asn 725 73’0 735 Leu <210> 35 <211> 2211 <212> DNA <213> Adeno-associated virus <400> 35 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acttgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata accacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 ggaaagaaga gaccggtaga gcaatcaccc caggaaccag actcctcttc gggcatcggc 480 aagaaaggcc agcagcccgc gaaaaagaga ctcaactttg ggcagacagg cgactcagag 540 tcagtgcccg accctcaacc actcggagaa ccccccgcag ccccctctgg tgtgggatct 600 aatacaatgg ctgcaggcgg tggcgctcca atggcagaca ataacgaagg cgccgacgga 660 gtgggtaacg cctcaggaaa ttggcattgc gattccacat ggctgggcga cagagtcatc 720 accaccagca cccgaacctg ggccctcccc acctacaaca accacctcta caagcaaatc 780 tccagccaat cgggagcaag caccaacgac aacacctact tcggctacag caccccctgg 840 gggtattttg actttaacag attccactgc cacttctcac cacgtgactg gcagcgactc 900 atcaacaaca actggggatt ccggcccaag agactcaact tcaagctctt caacatccag 960 gtcaaggagg tcacgacgaa tgatggcacc acgaccatcg ccaataacct taccagcacg 1020 gttcaggtct ttacggactc ggaataccag ctcccgtacg tcctcggctc tgcgcaccag 1080 ggctgcctgc ctccgttccc ggcggacgtc ttcatgattc ctcagtacgg gtacctgact 1140 ctgaacaatg gcagtcaggc cgtgggccgt tcctccttct actgcctgga gtactttcct 1200 tctcaaatgc tgagaacggg caacaacttt gagttcagct acacgtttga ggacgtgcct 1260 tttcacagca gctacgcgca cagccaaagc ctggaccggc tgatgaaccc cctcatcgac 1320 footcctgt actacctgtc tcggactcag accacgagtg gtaccgcagg aaatcggacg 1380 ttgcaatttt ctcaggccgg gcctagtagc atggcgaatc aggccaaaaa ctggctaccc 1440 gggccctgct accggcagca acgcgtctcc aagacagcga atcaaaaataa caacagcaac 1500 tttgcctgga ccggtgccac caagtatcat ctgaatggca gagactctct ggtaaatccc 1560 ggtcccgcta tggcaaccca caaggacgacgac gaacaaat tttttccgat gagcggagtc 1620 ttaatatttg ggaacaggg agctgggaaat agcaacgtgg accttgacaa cgttatgata 1680 accagtgagg aagaaattaa aaccaccaac ccagtggcca cagaacagta cggcacggtg 1740 gccactaacc tgcaatcgtc aaacaccgct cctgctacag ggaccgtcaa cagtcaagga 1800 gccttacctg gcatggtctg gcagaaccgg gacgtgtacc tgcagggtcc tatctgggcc 1860 aagattcctc acacggacgg acactttcat ccctcgccgc tgatgggagg ctttggactg 1920 aaacacccgc ctcctcagat cctgattaag aatacacctg ttcccgcgaa tcctccaact 1980 accttcagtc cagctaagtt tgcgtcgttc atcacgcagt acagcaccgg acaggtcagc 2040 gtggaaattg aatgggagct gcagaaagaa aacagcaaac gctggaaccc agagattcaa 2100 tacacttcca actacaacaa atctacaaat gtggactttg ctgttgacac aaatggcgtt 2160 tattctgagc ctcgccccat cggcacccgt tacctcaccc gtaatctgta a 2211 <210> 36 <211> 738 <212> PRT <213> Adeno-associated virus <400> 36 Met Ala Ala Asp Gly Tyr Leu Pro Asp Trp Leu Glu Asp Asn Leu Ser[[ID=二十九]] 1 5 10 15 Glu Gly Ile Arg Glu Trp Trp Asp Leu Lys Pro Gly Ala Pro Lys Pro 20 25 30 Lys Ala Asn Gln Gln Lys Gln Asp Asp Gly Arg Gly Leu Val Leu Pro 35 40 45 Gly Tyr Lys Tyr Leu Gly Pro Phe Asn Gly Leu Asp Lys Gly Glu Pro 50 55 60 Val Asn Ala Ala Asp Ala Ala Ala Leu Glu His Asp Lys Ala Tyr Asp 65 70 75 80 Gln Gln Leu Lys Ala Gly Asp Asn Pro Tyr Leu Arg Tyr Asn His Ala 85 90 95 Asp Ala Glu Phe Gln Glu Arg Leu Gln Glu Asp Thr Ser Phe Gly Gly 100 105 110 Asn Leu Gly Arg Ala Val Phe Gln Ala Lys Lys Arg Val Leu Glu Pro 115 120 125 Leu Gly Leu Val Glu Glu Gly Ala Lys Thr Ala Pro Gly Lys Lys Arg 130 135 140 Pro Val Glu Pro Ser Pro Gln Arg Ser Pro Asp Ser Ser Thr Gly Ile 145 150 155 160 Gly Lys Lys Gly Gln Gln Pro Ala Lys Lys Arg Leu Asn Phe Gly Gln 165 170 175 Thr Gly Asp Ser Glu Ser Val Pro Asp Pro Gln Pro Ile Gly Glu Pro 180 185 190 Pro Ala Gly Pro Ser Gly Leu Gly Ser Gly Thr Met Ala Ala Gly Gly 195 200 205 Gly Ala Pro Met Ala Asp Asn Asn Glu Gly Ala Asp Gly Val Gly Ser 210 215 220 Ser Ser Gly Asn Trp His Cys Asp Ser Thr Trp Leu Gly Asp Arg Val 225 230 235 240 Ile Thr Thr Ser Thr Arg Thr Trp Ala Leu Pro Thr Tyr Asn Asn His 245 250 255 Leu Tyr Lys Gln Ile Ser Asn Gly Thr Ser Gly Gly Ser Thr Asn Asp 260 265 270 Asn Thr Tyr Phe Gly Tyr Ser Thr Pro Trp Gly Tyr Phe Asp Phe Asn 275 280 285 Arg Phe His Cys His Phe Ser Pro Arg Asp Trp Gln Arg Leu Ile Asn 290 295 300 Asn Asn Trp Gly Phe Arg Pro Lys Arg Leu Asn Phe Lys Leu Phe Asn 305 310 315 320 Ile Gln Val Lys Glu Val Thr Gln Asn Glu Gly Thr Lys Thr Ile Ala 325 330 335 Asn Asn Leu Thr Ser Thr Ile Gln Val Phe Thr Asp Ser Glu Tyr Gln 340 345 350 Leu Pro Tyr Val Leu Gly Ser Ala His Gln Gly Cys Leu Pro Pro Phe 355 360 365 Pro Ala Asp Val Phe Met Ile Pro Gln Tyr Gly Tyr Leu Thr Leu Asn 370 375 380 Asn Gly Ser Gln Ala Val Gly Arg Ser Ser Phe Tyr Cys Leu Glu Tyr 385 390 395 400 Phe Pro Ser Gln Met Leu Arg Thr Gly Asn Asn Phe Glu Phe Ser Tyr 405 410 415 Gln Phe Glu Asp Val Pro Phe His Ser Ser Tyr Ala His Ser Gln Ser 420 425 430 Leu Asp Arg Leu Met Asn Pro Leu Ile Asp Gln Tyr Leu Tyr Tyr Leu 435 440 445 Ser Arg Thr Gln Ser Thr Gly Gly Thr Ala Gly Thr Gln Gln Leu Leu 450 455 460 Phe Ser Gln Ala Gly Pro Asn Asn Met Ser Ala Gln Ala Lys Asn Trp 465 470 475 480 Leu Pro Gly Pro Cys Tyr Arg Gln Gln Arg Val Ser Thr Thr Leu Ser 485 490 495 Gln Asn Asn Asn Ser Asn Phe Ala Trp Thr Gly Ala Thr Lys Tyr His 500 505 510 Leu Asn Gly Arg Asp Ser Leu Val Asn Pro Gly Val Ala Met Ala Thr 515 520 525 His Lys Asp Asp Glu Glu Arg Phe Phe Pro Ser Ser Gly Val Leu Met 530 535 540 Phe Gly Lys Gln Gly Ala Gly Lys Asp Asn Val Asp Tyr Ser Ser Val 545 550 555 560 Met Leu Thr Ser Glu Glu Glu Ile Lys Thr Thr Asn Pro Val Ala Thr 565 570 575 Glu Gln Tyr Gly Val Val Ala Asp Asn Leu Gln Gln Gln Asn Ala Ala 580 585 590 Pro Ile Val Gly Ala Val Asn Ser Gln Gly Ala Leu Pro Gly Met Val 595 600 605 Trp Gln Asn Arg Asp Val Tyr Leu Gln Gly Pro Ile Trp Ala Lys Ile 610 615 620 Pro His Thr Asp Gly Asn Phe His Pro Ser Pro Leu Met Gly Gly Phe 625 630 635 640 Gly Leu Lys His Pro Pro Pro Gln Ile Leu Ile Lys Asn Thr Pro Val 645 650 655 Pro Ala Asp Pro Pro Thr Thr Phe Ser Gln Ala Lys Leu Ala Ser Phe 660 665 670 Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val Glu Ile Glu Trp Glu 675 680 685 Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro Glu Ile Gln Tyr Thr 690 695 700 Ser Asn Tyr Tyr Lys Ser Thr Asn Val Asp Phe Ala Val Asn Thr Asp 705 710 715 720 Gly Thr Tyr Ser Glu Pro Arg Pro Ile Gly Thr Arg Tyr Leu Thr Arg 725 730 735 Asn Leu <210> 37 <211> twenty one <212> PRT <213> Adeno-associated virus <400> 37 Glu Ile Lys Ala Thr Asn Pro Val Ala Thr Glu Arg Phe Gly Thr Val 1 5 10 15 Ala Val Asn Phe Gln 20 <210> 38 <211> twenty one <212> PRT <213> Adeno-associated virus <400> 38 Glu Ile Lys Ala Thr Asn Pro Val Ala Thr Glu Arg Phe Gly Thr Val 1 5 10 15 Ala Val Asn Leu Gln 20 <210> 39 <211> twenty one <212> PRT <213> Adeno-associated virus <400> 39 Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr Gly Ser Val 1 5 10 15 Ser Thr Asn Leu Gln 20 <210> 40 <211> twenty one <212> PRT <213> Adeno-associated virus <400> 40 Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr Gly Thr Val 1 5 10 15 Ala Asn Asn Leu Gln 20 <210> 41 <211> twenty one <212> PRT <213> Adeno-associated virus <400> 41 Glu Ile Arg Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr Gly Thr Val 1 5 10 15 Ala Thr Asn Leu Gln 20 <210> 42 <211> twenty one <212> PRT <213> Adeno-associated virus <400> 42 Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr Gly Thr Val 1 5 10 15 Ala Thr Asn Leu Gln 20 <210> 43 <211> twenty one <212> PRT <213> Adeno-associated virus <220> <221> Variants <222> (12)..(12) <223> / Replace="Glu" <220> <221> misc_feature <222> (1)..(21) <223> / note="There is no preference for variant residues given in the sequence relative to those in the variant position annotation" <400> 43 Glu Ile Lys Thr Thr Asn Pro Val Ala Thr Glu Gln Tyr Gly Thr Val 1 5 10 15 Ala Thr Asn Leu Gln 20 <210> 44 <211> 6 <212> PRT <213> Artificial sequence <220> <221> source <223> / Note="Description of artificial sequence: synthetic 6xHis tag" <400> 44 His His His His His His 1 5

Claims

1. An adeno-associated virus (AAV) capsid polypeptide consisting of the amino acid sequence of SEQ ID NO:

7.

2. A nucleic acid molecule encoding the adeno-associated virus (AAV) capsid polypeptide according to claim 1.

3. A vector comprising the nucleic acid molecule of claim 2.

4. A host cell comprising the vector of claim 3.

5. A purified viral particle comprising the AAV capsid polypeptide of claim 1.

6. The purified viral particle of claim 5, further comprising a transgene.

7. Use of the virus particle of claim 6 in preparing a composition for gene transfer, wherein the composition is used in a method for gene transfer comprising: The viral particle of claim 6 is administered to a subject in need of gene transfer.

8. Use of the viral particle of claim 6 in the preparation of a composition for vaccination with a transgenic vaccine, wherein the composition is used in a method for vaccination with a transgenic vaccine, the method comprising: The viral particle of claim 6 is administered to a subject in need of vaccination.

9. Use of the AAV capsid polypeptide of claim 1 in the preparation of a composition for vaccinating a subject, wherein the composition is used in a method of vaccinating a subject, the method comprising: A target antigen operably linked to the AAV capsid polypeptide of claim 1 is administered to a subject in need of vaccination.

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