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 of 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 title of "Methods and Uses for Predicting Ancestral Viral Sequences". Technical Field

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

[0003] Avoiding and evading neutralizing or toxic immune responses against gene therapy vectors is a major challenge for all types of gene transfer vectors. Gene transfer to date has been most effectively achieved using vectors based on viruses that circulate in humans and animals (such as adenoviruses and adeno-associated viruses (AAV)). However, if a subject has been naturally infected with a virus, then subsequent treatment with a vector based on that virus results in increased safety risks and decreased efficiency of gene transfer due to cellular and humoral immune responses. Viral capsid antigens are mainly responsible for innate and / or adaptive immunity against viral particles; however, polypeptides encoded by viral genes can also be immunogenic. 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 virus (AAV); thus, the present disclosure describes predicted ancestral AAV sequences and AAV viral particles containing such ancestral AAV sequences. The present disclosure also describes a reduced seroprevalence exhibited by viral particles containing ancestral sequences relative to viral particles containing contemporary sequences.

[0005] In one aspect, the present disclosure includes adeno-associated virus (AAV) capsid polypeptides, e.g., synthetic and / or artificial AAV capsid polypeptides, 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 a viral particle comprising the AAV capsid polypeptide exhibits a lower seropositivity rate than an AAV2 capsid polypeptide or a viral particle comprising the AAV2 capsid polypeptide, and the AAV capsid polypeptide or a viral particle comprising the AAV capsid polypeptide exhibits a seropositivity rate that is approximately the same as or lower than that of an AAV8 capsid polypeptide or a viral particle comprising the AAV8 capsid polypeptide. In some embodiments, the AAV capsid polypeptide or a viral particle comprising the AAV capsid polypeptide is neutralized by human serum to a lesser extent than an AAV2 capsid polypeptide or a viral particle comprising the AAV2 capsid polypeptide, and the AAV capsid polypeptide or a viral particle comprising the AAV capsid polypeptide is neutralized by human serum to a similar or lower extent than an AAV8 capsid polypeptide or a viral particle 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 NO: 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 NO: 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 the AAV capsid polypeptides described herein. In some embodiments, the viral particles comprise 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 NO: 19, 20, 21, 22, 23, 24, 25, and 26. In some embodiments, the AAV capsid polypeptide or a virus particle comprising the AAV capsid polypeptide exhibits a lower seropositivity rate than an AAV2 capsid polypeptide or a virus particle comprising the AAV2 capsid polypeptide, and the AAV capsid polypeptide or a virus particle comprising the AAV capsid polypeptide exhibits an approximately the same or lower seropositivity rate as an AAV8 capsid polypeptide or a virus particle comprising the AAV8 capsid polypeptide. In some embodiments, the AAV capsid polypeptide or a virus particle comprising the AAV capsid polypeptide is neutralized by human serum to a lesser extent than an AAV2 capsid polypeptide or a virus particle comprising the AAV2 capsid polypeptide, and the AAV capsid polypeptide or a virus particle comprising the AAV capsid polypeptide is neutralized by human serum to a similar or lesser extent than an AAV8 capsid polypeptide or a virus particle 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 a nucleic acid sequence as described herein. In one embodiment, the present disclosure provides a nucleic acid molecule encoding an adeno-associated virus (AAV) capsid polypeptide, wherein the nucleic acid molecule has at least 95% (e.g., 97, 98, 99, or 100%) sequence identity to the nucleic acids as shown herein. The present disclosure also provides a vector comprising such a nucleic acid molecule, and also provides a host cell comprising such a vector.

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

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

[0012] In one aspect, the present disclosure provides a method 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 seropositivity rate than the AAV2 capsid polypeptide. In some embodiments, the AAV capsid polypeptide exhibits a seropositivity rate that is approximately the same or lower than the AAV8 capsid polypeptide. In some embodiments, the AAV capsid polypeptide is neutralized by human serum to a lesser extent than the AAV2 capsid polypeptide, and the AAV capsid polypeptide is neutralized by human serum to a similar or lesser extent than the AAV8 capsid polypeptide.

[0013] In another aspect, the present disclosure provides a computer method (insilico 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 of the plurality of contemporary viruses or portions thereof; estimating the evolutionary probability of a particular nucleotide or amino acid residue at each position in the sequence at a phylogenetic node of the predicted ancestral phylogeny and predicting the sequence of the ancestral virus or a portion thereof based on the estimated probabilities at each position.

[0014] In some embodiments, one or more or all of the steps are performed using a computer processor. In some embodiments, the MSA algorithm uses phylogenetic information to predict whether a gap in the alignment is the result of a deletion or an insertion. In some embodiments, the MSA algorithm is the Probabilistic Alignment Kit (PRANK). In some embodiments, the model used for modeling evolution is selected using the Aikake Information Criterion (AIC). In some embodiments, the predicted ancestral phylogeny is obtained using the JTT model and the Gamma distribution model (“+G”) and the calculation of the frequencies of πi (“+F”). In some embodiments, the step of modeling evolution is performed using the JTT+G+F model. In some embodiments, the method includes synthesizing the ancestral virus or a portion thereof based on the predicted sequence. In some embodiments, the method includes assembling the ancestral virus or a portion thereof into an ancestral virus particle.

[0015] In some embodiments, the method further comprises screening the ancestral viral particles for at least one of the following: (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, the ancestral viral particles are neutralized by human serum to a lesser extent than viral particles assembled from at least one of a plurality of contemporary viruses or portions thereof. 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, herpes simplex virus (HSV), vaccinia virus, pox virus, influenza virus, respiratory syncytial virus, parainfluenza virus, and foamy virus.

[0016] Accordingly, the present disclosure provides ancestral viruses or portions thereof that exhibit a reduced susceptibility to pre-existing immunity in the contemporary population compared to contemporary viruses or portions thereof. Generally, the reduced susceptibility to pre-existing immunity exhibited by ancestral viruses or portions thereof in the current population is reflected as a reduced susceptibility to neutralizing antibodies.

[0017] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the methods and compositions of matter belong. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the methods and compositions of matter, the following describes suitable methods and materials. In addition, the materials, methods, and examples are illustrative only and 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 NO: 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 a viral particle comprising the AAV capsid polypeptide exhibits a lower seroprevalence than an AAV2 capsid polypeptide or a viral particle comprising an AAV2 capsid polypeptide, and wherein the AAV capsid polypeptide or a viral particle comprising the AAV capsid polypeptide exhibits the same or lower seroprevalence than an AAV8 capsid polypeptide or a viral particle comprising an AAV8 capsid polypeptide.

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

[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, which has 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, which comprises the AAV capsid polypeptide of any one of embodiments 1-5.

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

[0029] 11. An adeno-associated virus (AAV) capsid polypeptide, which has at least 95% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 19, 20, 21, 22, 23, 24, 25, and 26.

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

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

[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 with 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 with 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, which further comprises a transgene.

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

[0038] administering the viral particle of embodiment 10 or embodiment 18 to a subject in need of gene transfer or immunization, wherein the viral particle exhibits a lower seropositivity rate than an AAV2 viral particle.

[0039] 20. The method of embodiment 19, wherein the viral particle exhibits a substantially same or lower seropositivity rate as an AAV8 viral particle.

[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 of vaccinating a subject, the method comprising

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

[0043] 23. The method of embodiment 22, wherein the AAV capsid polypeptide exhibits a seropositivity rate that is approximately the same as or lower than that of an 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 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.

[0045] 25. A computer method (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 a plurality of contemporary viruses or portions thereof;

[0047] aligning the sequences using a 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 specific nucleotide or amino acid residue at each position in the sequences at phylogenetic nodes of the predicted ancestral phylogeny, and

[0050] predicting the sequence of the ancestral virus or a portion thereof based on the estimated probabilities 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 the Probabilistic Alignment Kit (PRANK).

[0054] 29. The method of any one of embodiments 25 to 28, wherein a model for evolutionary modeling 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 the JTT model, the gamma distribution model (“+G”), and the calculation of the frequencies of πi (“+F”).

[0056] 31. The method of embodiment 25, wherein the step of evolutionary modeling is performed using the 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 ancestral virus particles.

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

[0060] 35. The method of embodiment 34, wherein the ancestral virus particles exhibit a lower seropositivity than virus 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 virus particles are neutralized to a lesser extent by human serum than virus particles assembled from at least one of the plurality of contemporary viruses or portions thereof.

[0062] 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, herpes simplex virus (HSV), vaccinia virus, pox virus, influenza virus, respiratory syncytial virus, parainfluenzavirus, 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 drawings 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 responses.

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

[0066] Figure 3 together show a phylogenetic tree of AAV contemporary sequences generated using the methods described herein.

[0067] Figure 4 together show an alignment of ancestral AAV VP1 polypeptides.

[0068] Figure 5A and 5B together show an alignment of functional ancestral AAV VP1 polypeptides and contemporary AAV VP1 polypeptides.

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

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

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

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

[0073] Figure 10 A - D are images of experimental results showing that Anc80L65 is able to assemble and produce high - titer particles. Figure A shows that Anc80L65 can produce a vector yield comparable to AAV2; Figure B is a TEM image of virus particles including Anc80L65; Figure C shows virus particles including Anc80L65, and based on SDS - PAGE gels under denaturing conditions, the virus particles can produce AAV cap VP1, 2, and 3 proteins; and Figure D shows a Western blot of Anc80L65 using the AAV capsid antibody B1.

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

[0075] Figure 12 Figures A - D show that Anc80L64 has a minimal seropositivity rate in human populations using IVIg (Figure A) or sera from Belgian populations (Figure B), sera from Boston populations (Figure C), or sera from cynomolgus macaques (Figure D).

[0076] Figure 13A and 13B is a sequence identity matrix generated using MAFFT, showing the amino acid sequence alignment of the VP1 protein of ancestral vectors with the amino acid sequence of the VP1 protein of representative extant AAVs ( Figure 13A ), and the amino acid sequence alignment of the VP3 protein of ancestral vectors with the amino acid sequence of the VP3 protein of representative extant AAVs ( Figure 13B ).

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

[0078] Figure 15 Table showing the titer of each vector, taking the average and comparing it with the titer of AAV8.

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

[0080] Figure 17 Figure showing the experimental results in which the same cells from Figure 16 were lysed and luciferase expression was measured. As in Figure 16 , Anc126 was not titrated against other vectors, but the range was an MOI of 2.5E2 - 3.1E2 GC / cell.

[0081] Figure 18 Table showing the luminescence of cells transduced by each vector, taking the average and comparing it with the luminescence of AAV8.

[0082] Figure 19 Figure providing a summary of in vitro experiments to determine 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 the main determinant of the efficiency, specificity, host response, pharmacology, and lifespan of the gene transfer reaction. Currently, the most efficient and effective way to achieve gene transfer is by using a vector or vector system based on a virus that has become replication-deficient.

[0084] However, seroprevalence studies indicate that a substantial proportion of the global human population has been pre-exposed (e.g., through natural infection) to a large number of the viruses currently used in gene transfer and thus 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, e.g., Calcedo et al., (2009, J. Infect. Dis., 199:381-90) and Boutin et al., (2010, Human Gene Ther., 21:704-12). Accordingly, the present disclosure is based on the recognition that ancestral viruses or portions thereof exhibit reduced susceptibility to pre-existing immunity (e.g., reduced susceptibility to neutralizing antibodies) in the current human population compared to contemporary viruses or portions thereof.

[0085] Figure 1 is a schematic diagram showing the relationship between ancestral and contemporary virus infections and ancestral and contemporary host immune responses. Figure 1 Shows how ancestral AAV can be refractory to contemporary pre-existing immunity. It is hypothesized that contemporary extant 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 extant viruses in terms of epitope composition.

[0086] The present disclosure provides methods for predicting the sequences of ancestral viruses or portions thereof. One or more ancestral virus sequences predicted using the methods described herein can be generated and assembled into virus particles. As demonstrated herein, virus particles assembled from predicted ancestral virus sequences can exhibit a lower, sometimes significantly lower, seroprevalence than current contemporary virus particles. Accordingly, 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 Virus Sequences

[0088] To predict an ancestral viral sequence, nucleotide or amino acid sequences are first compiled from multiple contemporary viruses or portions thereof. Although the methods described herein are exemplified with adeno-associated virus (AAV) capsid sequences, the same methods can be applied to other sequences from AAV (e.g., the entire genome, rep sequences, ITR sequences) or any other virus or portion thereof. Viruses other than AAV include, but are not limited to, adenovirus (AV), human immunodeficiency virus (HIV), retroviruses, lentiviruses, herpes simplex virus (HSV), measles virus, vaccinia virus, poxvirus, influenza virus, respiratory syncytial virus, parainfluenza virus, foamy virus, or any other virus for which pre-existing immunity is considered problematic.

[0089] Sequences from as few as two contemporary viruses or portions can be used; however, it is understood that a greater number of sequences of contemporary viruses or portions thereof are desirable in order to include as much of the landscape of modern 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, sequences from 100 or more contemporary viruses or portions thereof can be used.

[0090] Such sequences can be obtained, for example, from a number of public databases including, but not limited to, GenBank, UniProt, EMBL, the International Nucleotide Sequence Database Collaboration (INSDC), or the European Nucleotide Archive. Additionally / or, such sequences can be obtained from databases specific to a particular organism (e.g., an HIV database). Contemporary sequences can correspond to an 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 the viral capsid, replication proteins, or ITR sequences.

[0091] Next, the contemporary sequences are aligned using a multiple sequence alignment (MSA) algorithm. Figure 2FIG. (a) is a schematic diagram showing the alignment of multiple sequences. MSA algorithms are well known in the art and are generally designed to work with datasets of different sizes and different inputs (e.g., nucleic acids or proteins), and to align sequences in a particular way (e.g., dynamic programming, progressive, heuristic) and apply different scoring schemes in the alignment (e.g., matrix-based or consensus-based, e.g., minimum entropy, pair sum, similarity matrix, gap score). Well-known MSA algorithms include, for example, ClustalW (Thompson et al., 1994, Nuc. Acids Res., 22:4673-90), Kalign (Lassmann 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 an MSA algorithm for use in the methods described herein is the way in which the algorithm handles gaps in the alignment. A penalty value can be assigned to gaps in the sequence alignment, which may or may not depend on the gap size. In the present method, preferably, in contrast to the biased, non-phylogenetic treatment of gaps caused by, for example, insertions and / or deletions, the MSA algorithms used in the methods described herein apply phylogenetic information to predict whether a gap in the alignment is the result of a deletion or an insertion. Suitable methods for handling gaps in alignment and evolutionary analysis are described in Loytynoja and Goldman, 2008, Science, 320:1632-5, and commercially available algorithms that apply gaps to the alignment in a manner suitable for use in the methods described herein are the Probabilistic Alignment Kit (PRANK; Goldman Group Software; Loytynoja and Goldman, 2005, PNAS USA, 102:10557-62) and variants of the PRANK algorithm.

[0093] Then, an evolutionary model is applied to the resulting alignment to obtain a predicted ancestral phylogeny (see Figure 2(b)). There are many available evolutionary models in the art, each of which applies a slightly different matrix of substitution rates to amino acids. Algorithms for applying 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 D.C.), 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, for example, by considering that some positions are invariant (“+I”; Reeves, 1992, J. Mol. Evol., 35: 17-31), some positions experience changes 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), the constraints imposed on the evolutionary model by structure and function can be modeled themselves.

[0095] The Akaike 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 variants or combinations thereof can be used to evaluate the fitness of one or more evolutionary models. In addition, AIC, BIC, or variants or combinations thereof can be used to evaluate the relative importance of including one or more parameters (such as the constraints discussed above) in an 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 different from the AAV capsid polypeptide. However, those skilled in the art will also understand that different evolutionary models may be more appropriate depending on the dataset and fitness score.

[0097] Once an evolutionary model has been selected and its fitness determined, a phylogenetic tree of the viral sequence or a portion thereof can be constructed. Constructing phylogenetic trees is known in the art and typically uses the maximum likelihood method, 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). Those skilled in the art will understand that a balance between computational complexity and goodness of fit in models of amino acid substitution is desirable.

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

[0099] At any phylogenetic node of the phylogeny (e.g., an internal phylogenetic node), the sequence can be reconstructed by estimating the evolutionary probability of a specific nucleotide or amino acid residue at each position in the sequence (asFigure 2 (c)). A phylogenetic node is an intermediate evolutionary branch point within a predicted ancestral phylogeny. As used herein, "evolutionary probability" refers to the probability of the presence of a particular nucleotide or amino acid at a particular position based on an evolutionary model as contrasted with a model that does not account for evolutionary shifts such as codon selection. Any number of maximum likelihood methods can be used, including but not limited to 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) to evaluate exemplary models that account for the evolutionary probability of a particular nucleotide or amino acid residue at a particular position.

[0100] Based on the evaluated evolutionary probability of a particular nucleotide or amino acid residue at each position, a predicted sequence of an ancestral virus or a portion thereof can be assembled to form a complete or partial synthetic nucleic acid or polypeptide sequence. As needed, the likelihood of any residue being in a given state at a given node along the node can be calculated, and any positions along the sequence having a calculated posterior probability below a particular threshold can be identified ( Figure 2 (d)). In this way, an ancestral scaffold sequence can be generated that can include variations at those positions having a probability below a particular 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) to the ancestral scaffold sequence, and the ancestral sequence as described herein can be codon optimized for codon selection in mammals (e.g., humans).

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

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

[0104] Apply the methods described herein to adeno-associated virus (AAV) (described in detail in the following examples) using contemporary capsid sequences. AAV is widely regarded as a therapeutic gene transfer vector and a genetic vaccine vector, but exhibits a high seropositivity rate in the population. Using the methods described herein, a phylogenetic tree was assembled using contemporary AAV sequences (see Figure 3 ) and predicted ancestral scaffold sequences were obtained 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 probabilities and evolutionary modeling) and that is known not to exist in nature. As used herein, an ancestral scaffold sequence is different from a consensus sequence, which is typically constructed using the frequency of nucleotide 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 and 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 a virus or a portion thereof, a library of 2048 predicted ancestral sequences at the AAV Anc80 node was generated and, as described herein, shown to form live virus particles that exhibit a lower seropositivity rate than virus particles assembled with contemporary capsid polypeptides and, in some cases, a significantly lower seropositivity rate.

[0109] Methods for preparing ancestral virus particles

[0110] After a predicted ancestral sequence of a virus or a portion thereof has been obtained, actual nucleic acid molecules and / or polypeptides can be generated (e.g., synthesized). Methods for generating artificial nucleic acid molecules or polypeptides based on, for example, computer-derived sequences are known in the art and include, for example, chemical synthesis or recombinant cloning. Other methods for generating nucleic acid molecules or polypeptides are known in the art and are discussed in more detail below.

[0111] Once an ancestral polypeptide has been produced, or once an ancestral nucleic acid molecule has been produced and expressed to produce an ancestral polypeptide, the ancestral polypeptide can be assembled into ancestral virus particles using, for example, a packaging host cell. Components of the virus particles (e.g., rep sequences, cap sequences, inverted terminal repeat (ITR) sequences) can be introduced transiently or stably into the packaging host cell using one or more vectors as described herein. One or more components of the virus particles can be based on the predicted ancestral sequences as described herein, while the remaining components can be based on contemporary sequences. In some cases, the entire virus particle can be based on the predicted ancestral sequences.

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

[0113] Once assembled, the ancestral virus particles can be screened for, for example, the ability to replicate; gene transfer properties; receptor binding ability; and / or seropositivity in a population (e.g., the human population). Determining whether a virus particle can replicate is routine in the art and generally involves infecting host cells with a certain amount of virus particles and determining whether the number of virus particles increases over time. Determining whether a virus particle is capable of performing gene transfer is also routine in the art and generally involves infecting host cells with virus particles containing a transgene (e.g., a detectable transgene, such as a reporter gene, discussed in more detail below). After infection and clearance of the virus, the presence or absence of the transgene in the host cells can be evaluated. Determining whether a virus particle binds to its receptor is routine 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 involves using immunoassays to determine the prevalence of one or more antibodies in a sample (e.g., a blood sample) from a particular population of individuals. Seroprevalence is understood in the art to refer to the proportion of subjects in a population who are seropositive (i.e., have been exposed to a particular pathogen or immunogen), and is calculated as the number of subjects in the population who produce antibodies against a particular pathogen or immunogen 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, immunodot, Western blot, enzyme immunoassay (EIA), enzyme-linked immunosorbent assay (ELISA), or radioimmunoassay (RIA). As noted herein, ancestral viral particles exhibit a lower seroprevalence than contemporary viral particles (i.e., viral particles assembled using contemporary viral sequences or portions thereof). By way of example only, see Xu 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 determined the seroprevalence of a particular antibody in a given population.

[0115] As described herein, ancestral viral particles are neutralized to a lesser extent by the immune system of an individual (e.g., a patient) than contemporary viral particles. Several methods for determining the degree of neutralizing antibodies in a serum sample are available. For example, a neutralization antibody assay measures the titer of an antibody concentration in an experimental sample that neutralizes infection by 50% or more compared to a control sample 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 exemplary ancestral AAV capsid polypeptides herein, the seropositivity rate and / or degree of neutralization can be compared, for example, with 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 a relatively low seropositivity rate and resulting neutralization in the human population, while AAV2 capsid polypeptides or viral particles exhibit a relatively high seropositivity rate and resulting neutralization in the human population. Obviously, a particular seropositivity rate will depend on the population being examined and the immunological method used, but it has also been reported that AAV8 exhibits a seropositivity rate of about 22% up to about 38%, while AAV2 exhibits a seropositivity rate of about 43.5% up to about 72%. See, e.g., Boutin et al., 2010, “Prevalence of serum IgG and neutralizing factors against AAV types 1, 2, 5, 6, 8 and 9 in the healthy population: implications for gene therapy using AAV vectors,” Hum. Gene Ther., 21:704-12. See also, Calcedo et al., 2009, J. Infect. Dis., 199:381-90.

[0117] Predicted Adeno-Associated Virus (AAV) Ancestral Nucleic Acid and Polypeptide Sequences

[0118] Multiple different clones from a 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 understand that the nucleic acid sequences encoding each amino acid sequence can be readily determined.

[0119] In addition to the predicted ancestral capsid polypeptides having the sequences shown 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) with the predicted ancestral capsid polypeptides having the sequences shown 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) with nucleic acid molecules encoding ancestral capsid polypeptides (i.e., having at least 95% sequence identity).

[0120] In calculating the percent sequence identity, the two sequences are aligned and the number of identical matches of nucleotide or amino acid residues between the two sequences is determined. The number of identical matches is divided by the length of the alignment region (i.e., the number of aligned nucleotide or amino acid residues), and multiplied by 100 to obtain the percent sequence identity value. It should be understood that the length of the alignment region can be a part of one or both sequences up to the full-length size of the shortest sequence. It should also be understood that a single sequence can be aligned with more than one other sequence and thus can have different percent sequence identity values relative to each alignment region.

[0121] The alignment of two or more sequences to determine percent sequence identity can be carried out 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. BLAST searches can be performed to determine the percent sequence identity between a sequence (nucleic acid or amino acid) aligned using the algorithm of Altschul et al. and any other sequence or part thereof. BLASTN is the program for aligning and comparing identities between nucleic acid sequences, while BLASTP is the program for aligning and comparing identities between amino acid sequences. When using the BLAST program to calculate the percent identity between one sequence and another, the default parameters of the respective programs are typically used.

[0122] Representative alignments are shown in Figure 4 and Figure 5A and 5B . Figure 4shows the alignment of 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). The alignment shown in Figure 4 confirms predicted variations at each of 11 positions, as well as a single non-synonymous mutation at position 609E of Anc80L60 (SEQ ID NO: 21), which may be a cloning artifact. Figure 5A and 5B show the 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)). The alignment in Figure 5A and 5B shows that the ancestral AAV sequences have sequence identity between approximately 85% and 91% with the contemporary AAV sequences.

[0123] Vectors containing nucleic acid molecules encoding polypeptides are also provided. Vectors (including expression vectors) are commercially available or can be generated by recombinant techniques. A vector containing a nucleic acid molecule can have one or more expression elements operably linked to such nucleic acid molecule and can also include sequences such as those encoding selectable markers (e.g., antibiotic resistance genes), and / or those sequences useful in polypeptide purification (e.g., 6xHis tag). Expression elements include nucleic acid sequences that direct and regulate the expression of a nucleic acid coding sequence. An example of an expression element 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 a nucleic acid molecule. Expression elements can be of bacterial, yeast, insect, mammalian, or viral origin, and a vector can contain a combination of expression elements from different origins. As used herein, operably linked means that, relative to the coding sequence, the expression element is positioned in the vector in such a way as to direct or regulate the expression of the coding sequence.

[0124] A nucleic acid molecule, such as a nucleic acid molecule in a vector (e.g., an expression vector, a viral vector), can be introduced into a host cell. The term "host cell" refers not only to a particular cell that has received the introduction of a nucleic acid molecule, but also to the progeny or potential progeny of such a cell. Many suitable host cells are known to those of skill in the art; host cells can be prokaryotic cells (e.g., E. coli) or eukaryotic cells (e.g., yeast cells, insect cells, plant cells, mammalian cells). Representative 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, 293 cells, Saos, C2C12, L cells, HT1080, HepG2, and primary fibroblasts, hepatocytes, and myoblasts derived from mammals (including humans, monkeys, mice, rats, rabbits, and hamsters). 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 a polypeptide, "purified" means a polypeptide (i.e., a peptide or polypeptide) that has been separated or purified from the cellular components that naturally accompany it. Typically, a polypeptide is considered "purified" when it is at least 70% (e.g., at least 75%, 80%, 85%, 90%, 95% or 99%) by weight free of the polypeptides and naturally occurring molecules that are naturally associated with it. Since chemically synthesized polypeptides are inherently separated from the components that naturally accompany them, synthetic polypeptides are considered "purified", but may also be removed from the components used to synthesize the polypeptide (e.g., amino acid residues). With respect to a nucleic acid molecule, "isolated" means a nucleic acid molecule that is separated from other nucleic acid molecules that are normally associated with it in the genome. In addition, an isolated nucleic acid molecule may 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 a nucleic acid molecule in an expression vector or by chemical synthesis. Any suitable method, e.g., column chromatography, polyacrylamide gel electrophoresis, or HPLC analysis, can be used to measure the purity of a polypeptide. Similarly, nucleic acid molecules can be obtained (e.g., isolated) using conventional methods such as, but not limited to, recombinant nucleic acid techniques (e.g., restriction enzyme digestion and ligation) or polymerase chain reaction (PCR; see, e.g., PCR Primer: A Laboratory Manual, Dieffenbach & Dveksler, Eds., Cold Spring Harbor Laboratory Press, 1995). In addition, isolated nucleic acid molecules can be chemically synthesized.

[0127] Methods using an ancestral virus or a portion thereof

[0128] An ancestral virus or a portion thereof as described herein, particularly those ancestral viruses or portions thereof that exhibit a reduced seropositivity relative to a contemporary virus or a portion thereof, can be used in a number of research and / or therapeutic applications. For example, an ancestral virus or a portion thereof as described herein can be used in human or veterinary medicine for gene therapy (e.g., in a vector or vector system for gene transfer) or for vaccination (e.g., for antigen presentation). More specifically, an ancestral virus or a portion thereof as described herein can be used for gene addition, gene augmentation, heritable delivery of polypeptide therapeutics, genetic vaccination, gene silencing, genome editing, gene therapy, RNAi delivery, cDNA delivery, mRNA delivery, miRNA delivery, miRNA sponging, genetic immunization, optogenetic gene therapy, transgenesis, DNA vaccination or DNA immunization.

[0129] Host cells can be transduced or infected with an 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 an ancestral virus or a portion thereof in vitro are described herein; host cells that can be transduced or infected with an 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] An ancestral virus or portion as described herein can be modified to include a transgene (cis or trans to 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 that includes 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., cytokines and interleukins) or its receptor, an enzyme, an RNA (e.g., inhibitory RNA or catalytic RNA), or a target antigen (e.g., a carcinogenic 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 diseases, 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 (such as age-related macular degeneration), lipoprotein lipase deficiency, familial hypercholesterolemia, spinal muscular atrophy, Duchenne's 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, Huntington's disease, frontotemporal lobar degeneration, Usher syndrome, lactose intolerance, lipid storage disorder (such as Niemann-Pick type C), Batten disease, choroideremia, glycogen storage disease type II (Pompe disease), ataxia telangiectasia (Louis-Bar syndrome), congenital hypothyroidism, severe combined immunodeficiency (SCID), and / or amyotrophic lateral sclerosis (ALS).

[0132] The transgene can also be, for example, an immunogen 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] An ancestral virus or a portion thereof, which is typically suspended in a physiologically compatible carrier, can be administered to a subject (e.g., a human or a non-human mammal). Suitable carriers include saline (which can be formulated 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 a portion thereof is administered in an amount sufficient to transduce or infect cells and provide a sufficient level of gene transfer and expression to provide a therapeutic benefit without undue side effects. Conventional and pharmaceutically acceptable routes of administration include, but are not limited to, direct delivery to an organ, such as, for example, the liver or the lung, oral, intranasal, intratracheal, by inhalation, intravenous, intramuscular, intraocular, subcutaneous, intradermal, transmucosal, or by other routes of administration. Routes of administration can be combined as needed.

[0134] The dose of the ancestral virus or a portion thereof administered to a subject will depend primarily on factors such as the condition being treated, as well as the age, weight, and health of the subject. For example, a therapeutically effective dose of the ancestral virus or a portion thereof to be administered to a human subject is typically in the range from about 0.1 ml to about 10 ml of a solution containing a concentration of about 1×10 1 to 1×10 12 genomic 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 expression level of the transgene can be monitored to determine the frequency and / or amount of the dose. Dose 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 in order to modify or remove one or more immunogenic domains of a virus or a portion thereof.

[0136] In accordance with the present invention, conventional molecular biology, microbiology, biochemistry, and recombinant DNA techniques within the skill of the art can be employed. These techniques are well explained in the literature. The present invention will be further described in the following examples, which do not limit the scope of the methods and compositions described in the claims.

[0137] Examples

[0138] Example 1: In Silico Prediction of Ancestral Sequences

[0139] A set of 75 different amino acid sequences of AAV capsids was obtained from a number of public databases, including GenBank, and the sequences were aligned using the PRANK-MSA algorithm, version 121002, with the 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 models included in ProtTest3, i.e., JTT, LG, WAG, VT, CpRev, RtRev, Dayhoff, DCMut, FLU, Blosum62, VT, HIVb, MtArt, MtMam) under different conditions (e.g., those conditions included in ProTest3, i.e., “+I”, “+F”, “+G” and combinations thereof). The JTT model (Jones et al., 1992, Comp.Appl.Biosci., 8:275-82) with +G and +F (Yang, 1993, Mol.Biol.Evol., 10:1396-1401; and Cao et al., 1994, J.Mol.Evol., 39:519-27) was selected based on the Aikake Information Criterion (AIC; Hirotugu, 1974, IEEE Transactions on Automatic Control, 19:716-23) scores as implemented in ProtTest3.

[0141] The phylogeny of AAV evolution was constructed using PhyML (Guindon and Gascuel, 2003, Systematic Biology, 52:696-704). SeeFigure 3 Trees were generated using the JTT+F substitution model with 4 discrete substitution classes and an estimated Gamma shape parameter. The resulting trees were improved by Nearest Neighbor Interchange (NNI) and Subtree Pruning and Regrafting (SPR), and significance was evaluated using the "SH-Like" variant by bootstrapping and the approximate likelihood ratio test (aLRT; Anisimova and Gascuel, 2006, Systematic Biology, 55:539-52).

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

[0143] Since the reconstruction was done in a maximum likelihood manner, the likelihood of any residue at a given position at a given node could 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 the variants of interest in the library.

[0144] To finalize the sequence, additional utility must be encoded to select codons. A script was written to derive codons similar to those of another AAV sequence (AVVRh10, which has approximately 92% sequence identity with the Anc80 scaffold sequence) and a new algorithm was applied to replace codons where there were sequence mismatches based on a 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] Align Pi with Pt using Needleman-Wunsch with the Blosum62 table for scoring. A new nucleotide sequence, Ni, is generated by stepping through the protein alignment using the corresponding codons from Nt.

[0147] where the amino acid in Pt exactly matches the amino acid in Pi,

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

[0149] a gap where there is a gap in Pi aligned with the amino acid in Pt, and

[0150] the nucleotide that most frequently occurs in Nt (encoding the given amino acid) where there is an amino acid in Pi aligned with a gap in Pt.

[0151] In addition, two single nucleotide changes were also generated to eliminate the transcription of an assembly-activating protein (AAP) that is encoded out-of-frame within the AAV capsid gene in wild-type AAV. Since the encoding of AAP (contemporary or ancestral) is not part of this reconstruction, the expression of AAP 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 the ancestral AAV VP1 sequence

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

[0154] Numerous predicted ancestral AAV capsid sequences were cloned. The ancestral capsid library was transferred to a rep-cap expression plasmid, enabling the formation of viral particles in transient transfection. To maintain appropriate expression levels and splicing of VP1, VP2, and VP3, the library cap gene was cloned by cutting HindIII at the 5’ of cap located within the rep coding sequence and SpeI engineered between the cap stop codon and the polyadenylation signal. Thus, to clone the ancestral capsids into a more conventional “REP / CAP” construct, the transfer plasmid was digested with HindIII and SpeI, gel purified, and ligated into the similarly digested rep / cap plasmid.

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

[0156] Example 3: Virus Titration

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

[0158] The cell culture suspension was lysed by three consecutive freeze-thaw cycles. Thereafter, the lysate was clarified by centrifugation and treated with an enzyme (Benzonase TM ) under conditions for thorough DNA digestion to digest any DNA present outside the viral particles. The AAV preparation was diluted to fall within the linear measurement range of a control DNA template, in this case, a linearized plasmid with the same TaqMan TM primer and probe binding sequences compared to the vector genome. TaqMan TM PCR was performed with primers and probes annealed to the selected viral vector genome. The titer was calculated based on TaqMan TM measurement in 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 > Capsidless control <![CDATA[5.23x10 5 > <![CDATA[7.25x10 5 >

[0161] Small-scale vector production results on ancestrally reconstructed AAV capsid particles showed yields similar to AAV2 but reduced relative to AAV8 (both 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 cell seeds were transduced with each viral vector. Except for 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 quantifying bioluminescence in a 96-well plate reader. After quantification, a heatmap of luciferase expression in four cascading 96-well plates was generated (excluding the control columns in each plate). Due to the large number of insertions, deletions, and conversions associated with the high-throughput vector production process, many vectors were non-functional. For the purposes of this article, only functional viruses in this assay (i.e., capable of transducing HEK293 cells and expressing the transgene) were further evaluated.

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

[0166] Example 5 - In vivo retinal transduction

[0167] Retinal transduction was performed to determine whether ancestral AAV vectors are able to target murine retinal cells in vivo.

[0168] With 2×10 8Three different ancestral AAVs (Anc80L27, Anc80L44, and Anc80L65) and a contemporary AAV (AAV2 / 8) of genomic copies (GC) transduce the murine eye, all of which include the eGFP-encoding transgene. For transduction, each AAV vector was delivered subretinally by surgically creating a space between the photoreceptors and the retinal pigment epithelium layer via delivery of a vector bolus with an injection device. The vector bolus was left in the subretinal space and the retinal detachment resolved over time. By using Tropicamide TM The animal retinas after pupil dilation were non-invasively monitored for GFP expression by fundus photography. All presented retinas demonstrated varying degrees of successful targeting of the ancestral AAVs to the retina.

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

[0170] Example 6: Neutralizing Antibody Assay

[0171] A neutralizing antibody assay was performed to evaluate whether ancestral AAV viruses are more resistant to antibody neutralization than contemporary AAV viruses. The neutralizing antibody assay measures the antibody concentration (or the titer of the antibody concentration in the experimental sample) that neutralizes 50% or more of the infection 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 co-incubated with 10 4 MOI of the ancestral AAV virus, Anc80L65 and the contemporary AAV virus, AAV2 / 8, at 37 °C for approximately 30 minutes. Each virus includes the luciferase gene. Then the mixed vector and antibody samples were transduced into HEK293 cells. For these experiments, the antibody sample used was intravenous immunoglobulin (IVIG), from over 1000 blood donors (commercially available, e.g., Gammagard TM (Baxter Healthcare; Deerfield, IL) or Gamunex TMPooled IgG extracted from plasma of (Grifols; Los Angeles, CA)). At 48 hours after transduction initiation, cells were assayed by bioluminescence to detect luciferase. Neutralizing antibody titers were determined by identifying the dilution of samples that achieved 50% or greater neutralization (sample transduction / transduction of control virus in the absence of sample).

[0173] As Figure 8 shown, to reduce the transduction efficiency of the ancestral AAV virus Anc80L65 below 50% of the IVIG-free control (dashed line), significantly higher concentrations of IVIG were required compared to the contemporary AAV virus AAV2 / 8. These results demonstrate a higher resistance of the ancestral AAV virus to IVIG neutralization compared to contemporary AAV viruses.

[0174] Example 7: Characterization of Anc80

[0175] According to the methods described herein, the most likely Anc80 sequence (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). A Anc80 probability library was also synthesized by a commercial company using the sequences described herein and subcloned into an expression vector.

[0176] In a combined assay, the vector yield and infectivity of the Anc80 library were evaluated by cloning. In the screening, Anc80L65 (SEQ ID NO: 23) and various other variants were further characterized.

[0177] The Anc80 library and Anc80L65 were compared in terms of sequence differences ( Figure 9 ; % from the diagonal upwards, amino acid differences below). Using NCBI-BLAST, the most closely related publicly available sequence to Anc80L65 is rh10 (GenBank accession number AAO88201.1).

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

[0179] Briefly, small-scale production of AAV2 / 8, AAV2 / 2, AAV2 / Anc80L27, AAV2 / Anc80L44, and AAV2 / Anc80L65 vectors containing the reporter construct, which consists of eGFP and firefly luciferase under the control of the CMV promoter, was performed. Then, the titers of these small-scale preparations of the virus were 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 the Anc80L65 capsid protein assembled into intact virus-like particles of appropriate size and conformation, micrographs were obtained using transmission electron microscopy (TEM). A large-scale purified Anc80-L065 preparation was loaded onto a polyvinyl formal coated copper grid and then stained with uranyl acetate. The micrographs revealed intact, hexagonal particles with diameters between 20 and 25 nm ( Figure 10 B).

[0181] To determine whether the synthetic 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 an SDS-PAGE gel (1E10 GC / well) under denaturing conditions. For each vector preparation, bands representing the viral capsid proteins VP1, VP2, and VP3 were clearly present ( Figure 10 C). Western blotting with the AAV capsid antibody B1 further confirmed that these bands represented the predicted proteins ( Figure 10 D).

[0182] In addition, Figure 11 it was shown that, relative to AAV2 and / or AAV8 controls, using GFP as a readout (Figure A) or luciferase (Figure B), Anc80L65 infected mammalian tissues and cells in vitro on HEK293 cells at an MOI of 10E4 GC / cell. After IV injection of the indicated AAV encoding the nuclear LacZ transgene (top row, Figure C), after direct intramuscular (IM) injection of the indicated AAV encoding GFP (middle row, Figure C), and after subretinal injection of the indicated AAV encoding GFP (bottom row, Figure C), Anc80L65 also effectively targeted the liver. 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 were generated that contained a bicistronic reporter construct that included eGFP and firefly luciferase under the control of the CMV promoter. Then, 96-well plates confluent with HEK293 cells were transduced with each vector at an MOI of 1E4 GC / cell (titer obtained by qPCR as described above). After 48 hours, fluorescence microscopy confirmed the presence of GFP in the transduced cells ( Figure 11 A). Then, the presence of luciferase in the cells was assayed ( Figure 11 B), which determined that the expression of luciferase in cells transduced with Anc80-derived vectors was between the expression in cells transduced with AAV8 (lower level of transduction) and cells transduced with AAV2 (higher level of transduction).

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

[0185] Then, vectors of the same serotype were obtained that contained 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, each vector at 1E10 GC was injected into the skeletal muscle of C57BL / 6 mice (5 mice per condition) after general anesthesia. Twenty-eight days after injection, the mice were sacrificed, the tissue was cryosectioned, and the presence of eGFP was evaluated by fluorescence confocal microscopy (blue is DAPI, green is EGFP). Representative images are shown in Figure 11 C, middle row. These experiments demonstrated that the Anc80L65 vector was capable of transducing murine skeletal muscle by intramuscular injection.

[0186] Vectors of the same serotype were obtained, and this time encapsidated constructs encoding only the eGFP transgene under the control of the CMV promoter. After general anesthesia, 2E9 particles were injected sub - retinally into C57BL / 6 mice. Twenty - eight days after injection, the mice were sacrificed, the eyes were collected, cryosectioned, and the presence of eGFP was evaluated 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 capable of transducing the murine retina at levels comparable to those of the AAV8 vector.

[0187] Figure 12 Experimental results are shown where the seropositivity of ancestral viral vectors was evaluated relative to existing AAV viral vectors. Using an in vitro neutralizing antibody assay, it was shown that Anc80L65 exhibits increased resistance to neutralization using IVIG (pooled medicinal serum from approximately 10,000 individuals) ( Figure 12 A). Additionally, in box - and - whisker plots, sera from Belgian individuals (n = 100; Figure 12 C) or sera from Boston individuals (n = 102; Figure 12 B) show reduced sensitivity (or increased resistance) to neutralizing Anc80L65 relative to AAV2,8, and a similar seropositivity relative to rh32.33 (a different AAV vector with a known minimal seropositivity but limited use as a gene therapy vector). Sera obtained from cynomolgus monkeys exhibit a similar increased resistance relative to AAV2,8 and rh32.33 ( Figure 12 D). These experiments are described in more detail in the following paragraphs.

[0188] Briefly, highly purified high - titer preparations of AAV2 / 8, AAV2 / 2, AAV2 / rh32.33, and AAV2 / Anc80L65 viral vectors were obtained, and the viral vectors were encapsidated to include the eGFP and firefly luciferase bicistronic transgenes under the control of the CMV promoter. These vectors were then incubated with two - fold serial dilutions of IVIG (10 mg, 5 mg, 2.5 mg, etc.) or without IVIG (1E9 GC for each condition). After incubation, the vectors were used to transduce HEK293 cells at 1E4 MOI per well (one dilution per well). Forty - eight hours later, the relative amount of luciferase was measured by luminescence assay. Transduction relative to the 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 12Similar experiments were conducted for B). For each serum sample, the neutralization titer was reported as the dilution at which the given vector reduced by 50% relative to the serum-free control. In Figure 12 B and C of, the neutralization titers were reported as box-and-whisker plots, which determined that the Anc80L65 vector had a lower prevalence in these two populations (Belgian or Boston populations) compared to AAV2 and AAV8, approaching the level of Rh32.33 (a different AAV vector with a known minimal seropositivity rate). Serum obtained from cynomolgus monkeys was evaluated in the same manner ( Figure 12 D), where it was found that Anc80L65 had a significantly lower seropositivity rate compared to AAV2, AAV8, and the Rh.32.33 vector.

[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 to the amino acid sequences of extant AAVs to determine the degree to which they differed (Figure 13). Each ancestral VP1 protein differed from the VP1 protein of a selected representative extant AAV by 3.6% - 9.3% ( Figure 13A ), while the ancestral VP3 protein differed by 4.2% - 9.4% ( Figure 13B ). Each of these capsids was subcloned into the 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 mid-scale preparation of plasmid DNA was generated.

[0191] Each of these plasmids was then used to generate AAV vectors that contained a reporter gene encoding both eGFP and firefly luciferase. These vectors were generated in small-scale triplicate as described previously. The crude preparations of the viruses were then titrated by qPCR and found to produce 2.71% to 183.1% of the viral particles relative to AAV8 ( Figure 14 and 15 ). These titers were then used to set up titer control experiments to evaluate relative infectivity. Anc126 did not have a titer control due to its significantly reduced production, and therefore, data on 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 1.9E3 GC / cell.

[0192] Sixty hours after transduction, GFP expression in the cells was evaluated by fluorescence microscopy. eGFP-positive cells were detected under each condition except for the negative control ( Figure 16)。This indicates that each of the predicted, synthesized, and cloned ancestral sequences was able to produce live, infectious viral particles. To obtain an idea of the relative levels of infectivity, a luciferase assay was also performed on the same cells. The results indicate that each of the ancestral vectors 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 relative transduction analysis because it did not have a titer control.

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

[0194] Example 9: Vector Immunoprophylaxis

[0195] In vectored immunoprophylaxis, a gene therapy vector such as AAV is used to deliver a transgene encoding a broad-spectrum neutralizing antibody against an infectious agent. 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 the antibody in their own cells, which means that a single administration has the potential to confer lifelong protection against the etiologic agent.

[0196] Example 10: Drug Delivery Vector

[0197] (ranibizumab) and AVASTIN Both (bevacizumab) are anti-angiogenic agents based on the same humanized murine monoclonal antibody against vascular endothelial growth factor A (VEGF-A). Although bevacizumab is a full 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). Since these two molecules are proteins, they can be encoded by DNA and produced in cells transduced with a vector containing the transgene, and are small enough to be packaged into an AAV vector.

[0198] Other embodiments

[0199] It should be understood that although the methods and substance compositions have been described in connection with multiple different aspects herein, the foregoing description of the multiple aspects is intended to illustrate and not limit the scope of the methods and substance compositions. Other aspects, advantages, and modifications fall 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 combination with the products of the disclosed methods and compositions, can be used to prepare the products of the disclosed methods and compositions, or are the 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 substitution of these compositions and methods may not be explicitly disclosed, each is specifically contemplated and described herein. For example, if a particular substance composition or particular method is disclosed and discussed and multiple compositions and methods are discussed, then unless there is an express contrary indication, each combination and substitution of the compositions and methods is expressly covered. Similarly, any subgroup or combination of these is also expressly covered and disclosed.

[0201] Appendix A

[0202]

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210]

[0211]

[0212]

[0213] Sequence Listing <110> Massachusetts Eye and Ear Infirmary <120> Method 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> Variant <222> (168)..(168) <223> / replacement="Arg" <220> <221> Variant <222> (204)..(204) <223> / replacement="Ser" <220> <221> Variant <222> (266)..(266) <223> / replacement="Gly" <220> <221> Variant <222> (311)..(311) <223> / Replacement = "Lys" <220> <221> Variant <222> (411)..(411) <223> / Replacement = "Gln" <220> <221> Variant <222> (460)..(460) <223> / Replacement = "Glu" <220> <221> Variant <222> (493)..(493) <223> / Replacement = "Thr" <220> <221> Variant <222> (562)..(562) <223> / Replacement = "Asn" <220> <221> Variant <222> (576)..(576) <223> / Replacement = "Glu" <220> <221> Variant <222> (587)..(587) <223> / Replacement = "Ala" <220> <221> Variant <222> (609)..(609) <223> / Replacement = "Asp" <220> <221> misc_feature <222> (1)..(736) <223> / Note = "The variant residues given in the sequence have no preference over those in the annotation for the variant positions" <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> Mutation <222> (502)..(504) <223> / Replacement = "aaa" <220> <221> Mutation <222> (610)..(612) <223> / Replacement = "agc" <220> <221> Mutation <222> (796)..(798) <223> / Replacement = "ggc" <220> <221> Mutation <222> (931)..(933) <223> / Replacement = "aag" <220> <221> Mutation <222> (1231)..(1233) <223> / Replacement = "cag" <220> <221> Mutation <222> (1378)..(1380) <223> / Replacement = "gag" <220> <221> Mutation <222> (1477)..(1479) <223> / Replacement = "acc" <220> <221> Mutation <222> (1684)..(1686) <223> / Replacement = "aac" <220> <221> Mutation <222> (1726)..(1728) <223> / Replacement = "gag" <220> <221> Mutation <222> (1759)..(1761) <223> / Replacement = "gcc" <220> <221> Mutation <222> (1825)..(1827) <223> / Replacement = "gac" <220> <221> misc_feature <222> (1)..(2208) <223> / Note = "The mutated nucleotides given in the sequence have no preference over those in the mutation 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 cagtacctgt actacctgtc tcggactcag accacgagtg gtaccgcagg aaatcggacg 1380 ttgcaatttt ctcaggccgg gcctagtagc atggcgaatc aggccaaaaa ctggctaccc 1440 gggccctgct accggcagca acgcgtctcc aagacagcga atcaaaataa caacagcaac 1500 tttgcctgga ccggtgccac caagtatcat ctgaatggca gagactctct ggtaaatccc 1560 ggtcccgcta tggcaaccca caaggacgac gaagacaaat tttttccgat gagcggagtc 1620 ttaatatttg ggaaacaggg agctggaaat 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 gtaatctg 2208 <210> 3 <211> 737 <212> PRT <213> Adeno-associated virus <220> <221> Variant <222> (157)..(157) <223> / Replacement = "Ser" <220> <221> Variant <222> (168)..(168) <223> / Replacement = "Arg" <220> <221> Variant <222> (262)..(262) <223> / Replacement = "Ser" <220> <221> Variant <222> (263)..(263) <223> / Replacement = "His" <220> <221> Variant <222> (312)..(312) <223> / Replacement = "Lys" <220> <221> Variant <222> (412)..(412) <223> / Replacement = "Gln" <220> <221> Variant <222> (460)..(460) <223> / Replacement = "Gln" <220> <221> Variant <222> (461)..(461) <223> / Replacement = "Glu" <220> <221> Variant <222> (552)..(552) <223> / Replacement = "Ser" <220> <221> Variant <222> (556)..(556) <223> / replacement="Tyr" <220> <221> variant <222> (557)..(557) <223> / replacement="Ser" <220> <221> variant <222> (563)..(563) <223> / replacement="Asn" <220> <221> variant <222> (580)..(580) <223> / replacement="Ile" <220> <221> variant <222> (588)..(588) <223> / replacement="Ser" <220> <221> variant <222> (664)..(664) <223> / replacement="Thr" <220> <221> misc_feature <222> (1)..(737) <223> / note="The variant residues given in the sequence have no preference over those in the annotation for the variant positions" <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> Mutation <222> (469)..(471) <223> / Replacement = "agc" <220> <221> Mutation <222> (502)..(504) <223> / Replacement = "aag" <220> <221> Mutation <222> (784)..(786) <223> / Replacement = "agt" <220> <221> Mutation <222> (787)..(789) <223> / Replacement = "cac" <220> <221> Mutation <222> (934)..(936) <223> / Replacement = "aag" <220> <221> Mutation <222> (1234)..(1236) <223> / replacement="cag" <220> <221> Mutation <222> (1378)..(1380) <223> / replacement="cag" <220> <221> Mutation <222> (1381)..(1383) <223> / replacement="gag" <220> <221> Mutation <222> (1654)..(1656) <223> / replacement="agc" <220> <221> Mutation <222> (1666)..(1668) <223> / replacement="tac" <220> <221> Mutation <222> (1669)..(1671) <223> / replacement="agc" <220> <221> Mutation <222> (1687)..(1689) <223> / replacement="aac" <220> <221> Mutation <222> (1738)..(1740) <223> / replacement="atc" <220> <221> Mutation <222> (1762)..(1764) <223> / replacement="agc" <220> <221> Mutation <222> (1990)..(1992) <223> / replacement="acc" <220> <221> misc_feature <222> (1)..(2211) <223> / Note = "The variant nucleotides given in the sequence have no preference 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 ggaaagaaga gaccggtaga gcaatcaccc caggaaccag actcctctac gggcatcggc 480 aagaaaggcc agcagcccgc gaaaaagaga ctcaactttg ggcagactgg cgactcagag 540 tcagtgcccg accctcaacc actcggagaa ccccccgcag ccccctctgg tgtgggatct 600 aatacaatgg ctgcaggcgg tggcgctcca atggcagaca ataacgaagg cgccgacgga 660 gtgggtaatg cctcaggaaa ttggcattgc gattccacat ggctgggcga cagagtcatc 720 gtgggtaatg cctcaggaaa ttggcattgc gattccacat ggctgggcga cagagtcatc 720 accaccagca cccgaacctg ggccctcccc acctacaaca accacctcta caagcaaatc 780 accaccagca cccgaacctg ggccctcccc acctacaaca accacctcta caagcaaatc 780 tccaacagcc aatcgggagg aagcaccaac gacaacacct acttcggcta cagcaccccc 840 tccaacagcc aatcgggagg aagcaccaac gacaacacct acttcggcta cagcaccccc 840 tgggggtatt ttgactttaa cagattccac tgccacttct caccacgtga ctggcagcga 900 tgggggtatt ttgactttaa cagattccac tgccacttct caccacgtga ctggcagcga 900 ctcatcaaca acaactgggg attccggccc aagagactca acttcaagct cttcaacatc 960 ctcatcaaca acaactgggg attccggccc aagagactca acttcaagct cttcaacatc 960 caggtcaagg aggtcacgac gaatgatggc accacgacca tcgccaataa ccttaccagc 1020 caggtcaagg aggtcacgac gaatgatggc accacgacca tcgccaataa ccttaccagc 1020 acggttcagg tctttacgga ctcggaatac cagctcccgt acgtcctcgg ctctgcgcac 1080 acggttcagg tctttacgga ctcggaatac cagctcccgt acgtcctcgg ctctgcgcac 1080 cagggctgcc tgcctccgtt cccggcggac gtcttcatga ttcctcagta cgggtacctg 1140 cagggctgcc tgcctccgtt cccggcggac gtcttcatga ttcctcagta cgggtacctg 1140 actctgaaca atggcagtca ggccgtgggc cgttcctcct tctactgcct ggagtacttt 1200 actctgaaca atggcagtca ggccgtgggc cgttcctcct tctactgcct ggagtacttt 1200 ccttctcaaa tgctgagaac gggcaacaac tttgagttca gctacacgtt tgaggacgtg 1260 ccttctcaaa tgctgagaac gggcaacaac tttgagttca gctacacgtt tgaggacgtg 1260 ccttttcaca gcagctacgc gcacagccaa agcctggacc ggctgatgaa ccccctcatc 1320 ccttttcaca gcagctacgc gcacagccaa agcctggacc ggctgatgaa ccccctcatc 1320 gaccagtacc tgtactacct gtctcggact cagaccacgg gaggtaccgc aggaaatcgg 1380 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> Variant <222> (158)..(158) <223> / Replacement="Ser" <220> <221> Variant <222> (169)..(169) <223> / Replacement="Arg" <220> <221> Variant <222> (564)..(564) <223> / Replacement="Asn" <220> <221> misc_feature <222> (1)..(738) <223> / Note="The variant residues given in the sequence have no preference over those in the annotation of the variant positions" <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> Mutation <222> (472)..(474) <223> / Replacement = "agc" <220> <221> Mutation <222> (505)..(507) <223> / Replacement = "aga" <220> <221> Mutation <222> (1690)..(1692) <223> / Replacement = "aac" <220> <221> misc_feature <222> (1)..(2214) <223> / Note = "The mutated nucleotides given in the sequence have no preference relative to those in the mutation position annotation" <400> 6 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acctgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gagtggtggg acctgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata atcacgccga cgccgagttt 300 cagcagctca aagcgggtga caatccgtac ctgcggtata atcacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 ggaaagaaga gaccggtaga gcagtcacca cagcgtgagc ccgactcctc cacgggcatc 480 ggaaagaaga gaccggtaga gcagtcacca cagcgtgagc ccgactcctc cacgggcatc 480 ggcaagaaag gccagcagcc cgccaaaaag agactcaatt tcggtcagac tggcgactca 540 ggcaagaaag gccagcagcc cgccaaaaag agactcaatt tcggtcagac tggcgactca 540 gagtcagtcc ccgaccctca acctctcgga gaacctccag cagcgccctc tggtgtggga 600 gagtcagtcc ccgaccctca acctctcgga gaacctccag cagcgccctc tggtgtggga 600 tctaatacaa tggctgcagg cggtggcgca ccaatggcag acaataacga aggtgccgac 660 tctaatacaa tggctgcagg cggtggcgca ccaatggcag acaataacga aggtgccgac 660 ggagtgggta attcctcggg aaattggcat tgcgattcca catggctggg cgacagagtc 720 ggagtgggta attcctcggg aaattggcat tgcgattcca catggctggg cgacagagtc 720 atcaccacca gcacccgaac ctgggccctg cccacctaca acaaccacct ctacaagcaa 780 atcaccacca gcacccgaac ctgggccctg cccacctaca acaaccacct ctacaagcaa 780 atctccaacg ggacctcggg aggcagcacc aacgacaaca cctactttgg ctacagcacc 840 ccctgggggt 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 agcaactttg cctggactgg tgccaccaag tatcatctga acggcagaga ctctctggtg 1560 aatccgggcg tcgccatggc aacccacaag gacgacgagg accgcttctt cccatccagc 1620 aatccgggcg tcgccatggc aacccacaag gacgacgagg accgcttctt cccatccagc 1620 ggcgtcctca tatttggcaa gcagggagct ggaaatgaca acgtggacta tagcaacgtg 1680 ggcgtcctca tatttggcaa gcagggagct ggaaatgaca acgtggacta tagcaacgtg 1680 atgataacca gcgaggaaga aatcaagacc accaaccccg tggccacaga agagtatggc 1740 atgataacca gcgaggaaga aatcaagacc accaaccccg tggccacaga agagtatggc 1740 gtggtggcta ctaacctaca gtcggcaaac accgctcctc aaacggggac cgtcaacagc 1800 gtggtggcta ctaacctaca gtcggcaaac accgctcctc aaacggggac cgtcaacagc 1800 cagggagcct tacctggcat ggtctggcag aaccgggacg tgtacctgca gggtcctatt 1860 cagggagcct tacctggcat ggtctggcag aaccgggacg tgtacctgca gggtcctatt 1860 tgggccaaga ttcctcacac agatggcaac tttcacccgt ctcctttaat gggcggcttt 1920 tgggccaaga ttcctcacac agatggcaac tttcacccgt ctcctttaat gggcggcttt 1920 ggacttaaac atccgcctcc tcagatcctc atcaaaaaca ctcctgttcc tgcggatcct 1980 ggacttaaac atccgcctcc tcagatcctc atcaaaaaca ctcctgttcc tgcggatcct 1980 ccaacaacgt tcaaccaggc caagctgaat tctttcatca cgcagtacag caccggacaa 2040 ccaacaacgt tcaaccaggc caagctgaat tctttcatca cgcagtacag caccggacaa 2040 gtcagcgtgg agatcgagtg ggagctgcag aaggagaaca gcaagcgctg gaacccagag 2100 gtcagcgtgg agatcgagtg ggagctgcag aaggagaaca gcaagcgctg gaacccagag 2100 attcagtata cttccaacta ctacaaatct acaaatgtgg actttgctgt taatactgag 2160 attcagtata cttccaacta ctacaaatct acaaatgtgg actttgctgt taatactgag 2160 ggtgtttact ctgagcctcg ccccattggc actcgttacc tcacccgtaa tctg 2214 ggtgtttact ctgagcctcg ccccattggc actcgttacc tcacccgtaa tctg 2214 <210> 7 <210> 7 <211> 738 <212> PRT <213> Adeno-associated virus <220> <221> Variant <222> (158)..(158) <223> / Replacement = "Ser" <220> <221> Variant <222> (169)..(169) <223> / Replacement = "Lys" <220> <221> Variant <222> (315)..(315) <223> / Replacement = "Ser" <220> <221> Variant <222> (413)..(413) <223> / Replacement = "Glu" <220> <221> Variant <222> (472)..(472) <223> / Replacement = "Thr" or "Ser" <220> <221> Variant <222> (534)..(534) <223> / Replacement = "Glu" <220> <221> Variant <222> (542)..(542) <223> / Replacement = "Val" <220> <221> Variant <222> (595)..(595) <223> / Replacement = "Val" <220> <221> misc_feature <222> (1)..(738) <223> / Note = "The variant residues given in the sequence have no preference over those in the annotation for the variant positions" <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> Mutation <222> (472)..(474) <223> / Replacement = "agc" <220> <221> Mutation <222> (505)..(507) <223> / Replacement = "aag" <220> <221> Mutation <222> (943)..(945) <223> / Replacement = "agc" <220> <221> Mutation <222> (1237)..(1239) <223> / Replacement = "gaa" <220> <221> Mutation <222> (1414)..(1416) <223> / Replacement = "aac" or "agc" <220> <221> Mutation <222> (1600)..(1602) <223> / Replacement = "gag" <220> <221> Mutation <222> (1624)..(1626) <223> / Replacement = "gtc" <220> <221> Mutation <222> (1783)..(1785) <223> / Replacement = "gta" <220> <221> misc_feature <222> (1)..(2214) <223> / Note = "The mutated nucleotides given in the sequence have no preference relative to those in the mutation 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 ccctgggggt 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> Variant <222> (169)..(169) <223> / Replacement="Lys" <220> <221> Variant <222> (315)..(315) <223> / Replacement="Ser" <220> <221> Variant <222> (534)..(534) <223> / Replacement="Glu" <220> <221> Variant <222> (542)..(542) <223> / Replacement="Val" <220> <221> misc_feature <222> (1)..(738) <223> / Note="The variant residues given in the sequence have no preference over those in the annotation for the variant positions" <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> Variation <222> (505)..(507) <223> / Replacement = "aaa" <220> <221> Variation <222> (943)..(945) <223> / Replacement = "agc" <220> <221> Variation <222> (1600)..(1602) <223> / Replacement = "gag" <220> <221> Variation <222> (1624)..(1626) <223> / Replacement = "gtc" <220> <221> misc_feature <222> (1)..(2214) <223> / Note = "The variant nucleotides given in the sequence have no preference relative to those in the variant position annotation" <400> 10 atggctgccg atggttatct tccagattgg ctcgaggaca acctctctga gggcattcgc 60 gagtggtggg acctgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gagtggtggg acctgaaacc tggagccccg aaacccaaag ccaaccagca aaagcaggac 120 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 gacggccggg gtctggtgct tcctggctac aagtacctcg gacccttcaa cggactcgac 180 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 aagggggagc ccgtcaacgc ggcggacgca gcggccctcg agcacgacaa ggcctacgac 240 cagcagctca aagcgggtga caatccgtac ctgcggtata atcacgccga cgccgagttt 300 cagcagctca aagcgggtga caatccgtac ctgcggtata atcacgccga cgccgagttt 300 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 caggagcgtc tgcaagaaga tacgtctttt gggggcaacc tcgggcgagc agtcttccag 360 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 gccaagaagc gggttctcga acctctcggt ctggttgagg aaggcgctaa gacggctcct 420 ggaaagaaga gaccggtaga gccgtcacca cagcgttccc ccgactcctc cacgggcatc 480 ggaaagaaga gaccggtaga gccgtcacca cagcgttccc ccgactcctc cacgggcatc 480 ggcaagaaag gccagcagcc cgccagaaag agactcaatt tcggtcagac tggcgactca 540 ggcaagaaag gccagcagcc cgccagaaag agactcaatt tcggtcagac tggcgactca 540 gagtcagtcc ccgaccctca acctatcgga gaacctccag cagcgccctc tggtgtggga 600 gagtcagtcc ccgaccctca acctatcgga gaacctccag cagcgccctc tggtgtggga 600 tctggtacaa tggctgcagg cggtggcgca ccaatggcag acaataacga aggtgccgac 660 tctggtacaa tggctgcagg cggtggcgca ccaatggcag acaataacga aggtgccgac 660 ggagtgggta gttcctcggg aaattggcat tgcgattcca catggctggg cgacagagtc 720 ggagtgggta gttcctcggg aaattggcat tgcgattcca catggctggg cgacagagtc 720 atcaccacca gcacccgaac ctgggccctg cccacctaca acaaccacct ctacaagcaa 780 atcaccacca gcacccgaac ctgggccctg cccacctaca acaaccacct ctacaagcaa 780 atctccaacg ggacctcggg aggcagcacc aacgacaaca cctactttgg ctacagcacc 840 ccctgggggt 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> Variant <222> (471)..(471) <223> / replacement = "Asn" <220> <221> misc_feature <222> (1)..(738) <223> / note = "The variant residues given in the sequence have no preference over those in the annotation for the variant positions" <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> variant <222> (1411)..(1413) <223> / replacement="aat" <220> <221> misc_feature <222> (1)..(2214) <223> / note="The variant nucleotides given in the sequence have no preference over those in the annotation of the variant positions" <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 900 cgactcatca acaacaactg gggattccgg cccaagagac tcaacttcaa gctcttcaac 960 atccaggtca aggaggtcac gcagaatgaa ggcaccaaga ccatcgccaa taaccttacc 1020 agcacgattc aggtctttac ggactcggaa taccagctcc cgtacgtcct cggctctgcg 1080 caccagggct gcctgcctcc gttcccggcg gacgtcttca tgattcctca gtacgggtac 1140 ctgactctga acaatggcag tcaggccgtg ggccgttcct ccttctactg cctggagtac 1200 tttccttctc aaatgctgag aacgggcaac aactttgagt tcagctacac gtttgaggac 1260 gtgccttttc acagcagcta cgcgcacagc caaagcctgg accggctgat gaaccccctc 1320 atcgaccagt acctgtacta cctgtctcgg actcagtcca cgggaggtac cgcaggaact 1380 cagcagttgc tattttctca ggccgggcct agtaacatgt cggctcaggc caaaaactgg 1440 ctacccgggc cctgctaccg gcagcaacgc gtctccacga cactgtcgca aaataacaac 1500 agcaactttg cctggaccgg tgccaccaag tatcatctga atggcagaga ctctctggta 1560 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 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> Variant <222> (148)..(148) <223> / Replacement = "Gln" <220> <221> Variant <222> (169)..(169) <223> / Replacement = "Arg" <220> <221> Variant <222> (314)..(314) <223> / Replacement = "Asn" <220> <221> Variant <222> (466)..(466) <223> / Replacement = "His" <220> <221> Variant <222> (563)..(563) <223> / Replacement = "Ser" <220> <221> Variant <222> (580)..(580) <223> / Replacement = "Ile" <220> <221> Variant <222> (588)..(588) <223> / Replacement = "Ser" <220> <221> misc_feature <222> (1)..(737) <223> / Note = "The variant residues given in the sequence have no preference over those in the annotation for the variant positions" <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> Mutation <222> (442)..(444) <223> / Replacement="cag" <220> <221> Mutation <222> (505)..(507) <223> / Replacement="aga" <220> <221> Mutation <222> (940)..(942) <223> / Replacement="aac" <220> <221> Mutation <222> (1396)..(1398) <223> / Replacement="cac" <220> <221> Mutation <222> (1687)..(1689) <223> / Replacement="agt" <220> <221> Mutation <222> (1738)..(1740) <223> / Replacement="ata" <220> <221> Mutation <222> (1762)..(1764) <223> / Replacement="tct" <220> <221> misc_feature <222> (1)..(2211) <223> / Note = "The variant nucleotides given in the sequence have no preference 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 acctctcggt 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> Variant <222> (162)..(162) <223> / Replacement="Thr" <220> <221> Variant <222> (168)..(168) <223> / Replacement="Arg" <220> <221> Variant <222> (224)..(224) <223> / Replacement="Ser" <220> <221> Variant <222> (310)..(310) <223> / Replacement="Lys" <220> <221> Variant <222> (410)..(410) <223> / Replacement="Gln" <220> <221> Variant <222> (446)..(446) <223> / Replacement="Asn" <220> <221> Variant <222> (461)..(461) <223> / Replacement="Leu" <220> <221> Variant <222> (471)..(471) <223> / Replacement = "Ser" <220> <221> Variant <222> (708)..(708) <223> / Replacement = "Thr" <220> <221> misc_feature <222> (1)..(735) <223> / Note = "The variant residues given in the sequence have no preference relative to those in the mutation 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 Leucine Methionine Asparagine Proline Leucine Isoleucine Aspartic acid Glutamine Tyrosine Leucine Tyrosine Tyrosine Leucine Serine Arginine Threonine 435 440 445 Glutamine Threonine Threonine Serine Glycine Threonine Alanine Glutamine Asparagine Arginine Glutamic acid Leucine Glutamine Phenylalanine Serine Glutamine 450 455 460 Alanine Glycine Proline Serine Serine Methionine Alanine Asparagine Glutamine Alanine Lysine Asparagine Tryptophan Leucine Proline Glycine 465 470 475 480 Proline Cysteine Tyrosine Arginine Glutamine Glutamine Arginine Valine Serine Lysine Threonine Alanine Asparagine Aspartic acid Asparagine Asparagine 485 490 495 Asparagine Serine Asparagine Phenylalanine Alanine Tryptophan Threonine Glycine Alanine Threonine Lysine Tyrosine Histidine Leucine Asparagine Glycine 500 505 510 Arginine Aspartic acid Serine Leucine Valine Asparagine Proline Glycine Proline Alanine Methionine Alanine Serine Histidine Lysine Aspartic acid 515 520 525 Aspartic acid Glutamic acid Aspartic acid Lysine Phenylalanine Phenylalanine Proline Methionine Serine Glycine Valine Leucine Isoleucine Phenylalanine Glycine Lysine 530 535 540 Glutamine Glycine Alanine Glycine Alanine Serine Asparagine Valine Aspartic acid Leucine Aspartic acid Asparagine Valine Methionine Isoleucine Threonine 545 550 555 560 Aspartic acid Glutamic acid Glutamic acid Glutamic acid Isoleucine Lysine Threonine Threonine Asparagine Proline Valine Alanine Threonine Glutamic acid Glutamine Tyrosine 565 570 575 Glycine Threonine Valine Alanine Threonine Asparagine Leucine Glutamine Serine Serine Asparagine Threonine Alanine Proline Alanine Threonine 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> Mutation <222> (484)..(486) <223> / replacement="aca" <220> <221> Mutation <222> (502)..(504) <223> / replacement="aga" <220> <221> Mutation <222> (670)..(672) <223> / replacement="tcc" <220> <221> Mutation <222> (928)..(930) <223> / replacement="aaa" <220> <221> Mutation <222> (1228)..(1230) <223> / replacement="cag" <220> <221> Mutation <222> (1336)..(1338) <223> / replacement="aac" <220> <221> Mutation <222> (1381)..(1383) <223> / replacement="ctg" <220> <221> Mutation <222> (1411)..(1413) <223> / replacement="tct" <220> <221> Mutation <222> (2122)..(2124) <223> / replacement="acc" <220> <221> misc_feature <222> (1)..(2205) <223> / Note = "There is no preference for the mutant nucleotides given in the sequence relative to those in the mutant 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 gccaagaaga 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 ggctgggcga cagagtcatc 720 accaccagca cccgaacatg ggccttgccc acctataaca accacctcta caagcaaatc 780 tccagtcaat caggggccag caacgacaac cactacttcg gctacagcac cccctggggg 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> Variant <222> (42)..(42) <223> / Replacement="Ser" <220> <221> Variant <222> (168)..(168) <223> / Replacement="Lys" <220> <221> Variant <222> (310)..(310) <223> / Replacement="Arg" <220> <221> Variant <222> (410)..(410) <223> / Replacement="Gln" <220> <221> Variant <222> (446)..(446) <223> / Replacement="Arg" <220> <221> Variant <222> (461)..(461) <223> / Replacement="Leu" <220> <221> Variant <222> (471)..(471) <223> / Replacement="Ser" <220> <221> Variant <222> (475)..(475) <223> / Replacement="Arg" <220> <221> Variant <222> (504)..(504) <223> / Replacement="Ala" <220> <221> Variant <222> (539)..(539) <223> / Replacement="Asn" <220> <221> misc_feature <222> (1)..(735) <223> / Note="The variant residues given in the sequence have no preference relative to those in the mutation 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> Mutation <222> (124)..(126) <223> / Replacement = "agt" <220> <221> Mutation <222> (502)..(504) <223> / Replacement = "aaa" <220> <221> Mutation <222> (928)..(930) <223> / Replacement = "aga" <220> <221> Mutation <222> (1228)..(1230) <223> / Replacement = "cag" <220> <221> Mutation <222> (1336)..(1338) <223> / Replacement = "aga" <220> <221> Mutation <222> (1381)..(1383) <223> / Replacement = "ctc" <220> <221> Mutation <222> (1411)..(1413) <223> / Replacement = "tct" <220> <221> Mutation <222> (1423)..(1425) <223> / Replacement = "aga" <220> <221> Mutation <222> (1510)..(1512) <223> / Replacement="gcg" <220> <221> Mutation <222> (1615)..(1617) <223> / Replacement="gac" <220> <221> misc_feature <222> (1)..(2205) <223> / Note="The mutated nucleotides given in the sequence have no preference over those in the mutation 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 ggacgtcttc 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 gcttggacag 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 <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 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 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 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 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 <211> 736 <212> PRT <213> 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 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> 736 <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 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> Adeno-associated 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 705 710 715 720 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 <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 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 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 <210> 34 <211> 737 <212> PRT <213> 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 725 730 735 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 cagtacctgt actacctgtc tcggactcag accacgagtg gtaccgcagg aaatcggacg 1380 ttgcaatttt ctcaggccgg gcctagtagc atggcgaatc aggccaaaaa ctggctaccc 1440 gggccctgct accggcagca acgcgtctcc aagacagcga atcaaaataa caacagcaac 1500 tttgcctgga ccggtgccac caagtatcat ctgaatggca gagactctct ggtaaatccc 1560 ggtcccgcta tggcaaccca caaggacgac gaagacaaat tttttccgat gagcggagtc 1620 ttaatatttg ggaaacaggg agctggaaat 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 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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <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> 21 <212> PRT <213> Adeno-associated virus <220> <221> variant <222> (12)..(12) <223> / replacement="Glu" <220> <221> misc_feature <222> (1)..(21) <223> / note="The variant residues given in the sequence have no preference 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:

5.

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 transgene, wherein the composition is used in a method for vaccination with a transgene, 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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