Adeno-associated virus vector for GLUT1 expression and its use

Abstract

We provide gene therapy for GLUT1 deficiency syndrome. [Solution] This specification provides gene therapy for GLUT1 deficiency syndrome and related disorders using recombinant adeno-associated virus (rAAV) virions as vectors for expressing the GLUT1 protein or a functional variant thereof. The rAAV virions may use endothelial-specific promoters, such as the FLT-1 or Tie-1 promoter. The capsid may be the AAV6, AAV8, AAV9, AAVrh.74, or AAVrh.10 capsid or a functional variant thereof. Other promoters or capsids may be used. Furthermore, methods for therapy using rAAV virions intracerebral and / or intravenously, as well as other compositions and methods, are provided.

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims priority to U.S. Patent Application No. 63 / 061,726, filed on August 5, 2021, the entire content of which is incorporated herein by reference.

[0002] 1]Description of the Sequence Listing The sequence listing related to this application is provided in text format instead of a paper copy and is incorporated herein by reference. The name of the text file containing the sequence listing is ROPA_018_01WO_ST25.txt. The text file is approximately 190 KB, was created on August 3, 2021, and was electronically submitted via EFS - Web.

Background Art

[0003] Background Mutations in the SLC2A1 gene encoding glucose transporter 1 (GLUT1) are associated with a neurodevelopmental disorder called GLUT1 deficiency syndrome (GLUT1 DS). GLUT1 DS is an autosomal dominant disorder and often exists as a sporadic disease with de novo mutations that result in haploinsufficiency and symptomatic heterozygosity.

[0004] ]1] GLUT1 is an insulin-independent glucose transporter. Patients with classic GLUT1 DS, also known as Devibo disease, suffer from low brain glucose levels and exhibit a phenotype characterized by: early-onset seizures (median 12 months), developmental delay, acquired microcephaly (slowed head growth), complex motor impairment (spasticity, ataxia, dystonia), paroxysmal intraocular movements, and cerebrospinal fluid glucose deficiency, or low cerebrospinal fluid (CSF) glucose concentration. The clinical course of the disease highlights the importance of early treatment. Alter et al. J. Child Neurol. 30(2):160-169 (2015) (Non-patent Literature 1). GLUT1 is involved in endothelial cell function, including angiogenesis and maintenance of the blood-brain barrier (BBB). However, studies in haploinsufficiency mouse models have provided conflicting evidence regarding the role of GLUT1 in maintaining the physical integrity of the BBB. Endothelial cell lineage-specific knockout of GLUT1 reduces the availability of endothelial energy, decreases proliferation without affecting migration, and thereby delays developmental angiogenesis (Veys et al., Circ.Res.2020;127:466-482 (Non-patent Literature 2)), but its effect on restoring GLUT1 expression, particularly in endothelial cells, has not been tested.

[0005] Treatment strategies for the disease are outlined below, Tang et al. Ann. Clin. Trans. Neurol. 2019;6(9):1923-1932 (Non-Patent Literature 3). The current standard of care is ketogenic diet therapy, which increases levels of ketones in the blood to make them available to the brain as an alternative to glucose. Therapy with the triglyceride triheptanoin has been proposed as an alternative to ketogenic diet therapy. Gene therapy using adeno-associated virus (AAV) vectors has also been attempted. AAV9 vectors encoding GLUT1 under the control of a neuron-specific promoter (e.g., synapsin), targeting GLUT1 deficiency in neurons, have been tested in young postnatal mouse models. Other studies have used constitutive promoters (e.g., CMV promoter) or promoters of the endogenous GLUT1 gene. Various small molecules, including the anticonvulsant carbonic anhydrase inhibitor acetazolamide and others, have also been tested.

[0006] GLUT1 haploinsufficiency halts cerebral angiogenesis, resulting in a relatively small cerebral microvascular system, which may be related to glucose dependence in endothelial apical cells. However, Tang et al. observed that it has not yet been investigated whether low GLUT1 in endothelial cells induces this pathology. The GLUT1 protein is expressed in additional brain cells, including oligodendrocytes, microglia, and ependymal cells.

[0007] Addressing GLUT1 DS with gene therapy presents several challenges. Both the required degree of CNS coverage using the necessary vectors and the required level of GLUT1 treatment are highly unpredictable in order to achieve clinically meaningful efficacy.

[0008] There is an unmet need for the treatment of GLUT1 deficiency syndrome. The gene therapies presented herein address this need. [Prior art documents] [Non-patent literature]

[0009] [Non-Patent Document 1] Alter et al.J.Child Neurol.30(2):160-169(2015) [Non-Patent Document 2] Veys et al.,Circ.Res.2020;127:466-482 [Non-Patent Document 3] Tang et al.Ann.Clin.Trans.Neurol.2019;6(9):1923-1932 [Overview of the Initiative]

[0010] overview The present invention generally relates to gene therapy for neurological disorders or disabilities using adeno-associated virus (AAV)-based delivery of polynucleotides encoding GLUT1 or functional variants thereof.

[0011] GLUT1 deficiency syndrome (DS) is a neurodevelopmental disorder characterized by clinical symptoms rooted in a lack of proper neuronal function. However, this gene therapy, without being constrained by theory, can target endothelial cells involved in the development and induction of angiogenesis in the central nervous system (CNS). Direct delivery of AAV to the developing CNS vascular system, accompanied by subsequent expression of GLUT1 protein in endothelial apical cells, may promote vascular growth and formation throughout the CNS during a critical window of angiogenesis and neurodevelopment.

[0012] In one embodiment, the disclosure provides an expression cassette comprising a polynucleotide sequence encoding GLUT1 or a functional variant thereof, which is operably ligated to a promoter.

[0013] In some embodiments, the promoter is the endothelial promoter, optionally the Tie-1 promoter, the Tie-2 (TEK) promoter, the FLT-1 promoter, the FLK-1 (KDR) promoter, the ICAM-2 promoter, the VE-cadherin (CDH5) promoter, the VWF promoter, the ENG promoter, the PDGFB promoter, the ESM1 promoter, the APLN promoter, or the claudin-5 (Ple261) promoter, and the endothelial promoter is not the Glut1 promoter.

[0014] In some embodiments, the promoter is the FLT-1 promoter.

[0015] In some embodiments, the FLT-1 promoter is a human FLT-1 (hFTL-1) promoter.

[0016] In some embodiments, the hFTL-1 promoter shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 1.

[0017] In some embodiments, the promoter is a Tie-1 promoter.

[0018] In some embodiments, the Tie-1 promoter is the human Tie-1 (hTie-1) promoter.

[0019] In some embodiments, the hFTL-1 promoter shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 2.

[0020] In some embodiments, the promoter is a vascular endothelial-cadherin (VE-cadherin) promoter.

[0021] In some embodiments, the VE-cadherin promoter is a human VE-cadherin (hVE-cadherin) promoter.

[0022] In some embodiments, the hVE-cadherin promoter shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 3.

[0023] In some embodiments, the promoter is a ubiquitous promoter.

[0024] In some embodiments, the promoter is a CMV promoter.

[0025] In some embodiments, the promoter is a CAG promoter.

[0026] In some embodiments, the expression cassette includes a polyA signal, optionally including human growth hormone (hGH) polyA.

[0027] In some embodiments, the expression cassette includes a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), optionally WPRE(x).

[0028] In some embodiments, the expression cassette includes a 3'untranslated region (3'UTR) that includes a sequence sharing at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 4.

[0029] [[ID=3,0]]In some embodiments, the polynucleotide sequence encoding GLUT1 is an SLC2A1 polynucleotide.

[0030] In some embodiments, the SLC2A1 polynucleotide is a human SLC2A1 polynucleotide.

[0031] In some embodiments, the polynucleotide sequence encoding GLUT1 shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 5.

[0032] In some embodiments, the expression cassette is flanked by the 5' and 3' inverted terminal repeats (ITRs), optionally adjacent to the AAV2 ITR.

[0033] In some embodiments, the expression cassette shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with one of sequence numbers 8-16, sequence number 97, sequence number 99, and sequence number 101.

[0034] In another embodiment, the Disclosure provides a gene therapy vector comprising one of the expression cassettes of the Disclosure.

[0035] In some embodiments, the gene therapy vector is a recombinant adeno-associated virus (rAAV) vector.

[0036] In some embodiments, the rAAV vector is AAV6, AAV8, AAV9, or AAVrh.74, AAVrh.10 vector or a functional variant thereof.

[0037] In some embodiments, the rAAV vector is not an AAV2 vector.

[0038] In some embodiments, the rAAV vector contains a capsid protein that shares 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with any one of sequence numbers 76-82.

[0039] In another aspect, the Disclosure provides a method for treating and / or preventing a disease or disorder in a subject in need thereof, the method comprising the step of administering a vector described in any one of the Disclosures to a subject.

[0040] In some embodiments, the disease or disorder is a neurological disorder.

[0041] In some embodiments, the disease or disorder is glucose transporter 1 deficiency syndrome (GLUT1 DS) or Devibo disease.

[0042] In some embodiments, the vector is administered by intraventricular (ICV) injection.

[0043] In some embodiments, administration results in increased expression of the polynucleotide sequence encoding GLUT1 in the brain and / or increased glucose or lactate levels in CSF, at an optionally increased level compared to the reference rAAV vector, where the optional increase is at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or more.

[0044] In some embodiments, administration results in the expression of GLUT1 protein in the brain at selectively increased levels compared to the reference rAAV vector.

[0045] In some embodiments, the vector is administered in doses of 1E11 vector genome (vg), 1E12vg, 1E13, 1E14, 2E14, or 3E14.

[0046] In another embodiment, the present disclosure provides a method for expressing GLUT1 in cells, comprising the step of contacting the cells with one of the vectors of the present disclosure.

[0047] In some embodiments, the cells are endothelial cells.

[0048] In some embodiments, the endothelial cells are in vivo endothelial cells.

[0049] In some embodiments, the cells are neurons.

[0050] In some embodiments, the neurons are in vivo neurons.

[0051] In some embodiments, the method includes in vivo administration of the vector to a subject.

[0052] In further embodiments, the disclosure provides polynucleotides (e.g., vector genomes), pharmaceutical compositions, kits, and other compositions and methods.

[0053] [Invention 1001] An expression cassette comprising a polynucleotide sequence encoding GLUT1 or a functional variant thereof, operably ligated to a promoter. [Invention 1002] The expression cassette of the present invention 1001, wherein the promoter is an endothelial promoter, optionally a Tie-1 promoter, a Tie-2 (TEK) promoter, a FLT-1 promoter, a FLK-1 (KDR) promoter, an ICAM-2 promoter, a VE-cadherin (CDH5) promoter, a VWF promoter, an ENG promoter, a PDGFB promoter, an ESM1 promoter, an APLN promoter, or a claudin-5 (Ple261) promoter, provided that the endothelial promoter is not a Glut1 promoter. [Invention 1003] An expression cassette according to the present invention 1001 or 1002, wherein the promoter is an FLT-1 promoter. [Invention 1004] The expression cassette of the present invention 1003, wherein the FLT-1 promoter is a human FLT-1 (hFLT-1) promoter. [Invention 1005] An expression cassette of the present invention 1004, wherein the hFLT-1 promoter shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 1. [Invention 1006] An expression cassette according to the present invention 1001 or 1002, wherein the promoter is the Tie-1 promoter. [Invention 1007] The expression cassette of the present invention 1006, wherein the Tie-1 promoter is the human Tie-1 (hTie-1) promoter. [Invention 1008] An expression cassette of the present invention 1007, wherein the hTie-1 promoter shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 2. [Invention 1009] An expression cassette according to the present invention 1001 or 1002, wherein the promoter is a vascular endothelial-cadherin (VE-cadherin) promoter. [Invention 1010] The expression cassette of the present invention 1009, wherein the VE-cadherin promoter is a human VE-cadherin (hVE-cadherin) promoter. [Invention 1011] An expression cassette of the present invention 1010, wherein the hVE-cadherin promoter shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 3. [Invention 1012] The expression cassette of the present invention 1001, wherein the promoter is a ubiquitous promoter. [Invention 1013] An expression cassette according to the present invention 1001 or 1012, wherein the promoter is a CMV promoter. [Invention 1014] An expression cassette according to the present invention 1001 or 1012, wherein the promoter is a CAG promoter. [Invention 1015] An expression cassette according to any of the present invention 1001 to 1014, comprising a polyA signaling molecule and optionally containing human growth hormone (hGH) polyA. [Invention 1016] An expression cassette according to any of the inventions 1001 to 1015, comprising a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), optionally comprising WPRE(x). [Invention 1017] An expression cassette according to any of the inventions 1001 to 1016, comprising a 3' untranslated region (3'UTR) containing a sequence that shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with sequence number 4. [Invention 1018] An expression cassette according to any of the present invention 1001 to 1017, wherein the polynucleotide sequence encoding GLUT1 is the SLC2A1 polynucleotide. [Invention 1019] The expression cassette of the present invention 1018, wherein the SLC2A1 polynucleotide is human SLC2A1 polynucleotide. [Invention 1020] An expression cassette according to any of the invention 1017 to 1019, wherein the polynucleotide sequence encoding GLUT1 shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 5. [Invention 1021] An expression cassette of any of the Invention 1001-1020, adjacent to 5' and 3' inverted terminal repeats (ITRs), optionally to an AAV2 ITR, or optionally to an ITR that shares at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with SEQ ID NO: 6 or SEQ ID NO: 7. [Invention 1022] An expression cassette according to any of the invention 1001 to 1021, sharing at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identity with any one of sequence numbers 8 to 16, sequence number 97, sequence number 99, and sequence number 101. [Invention 1023] A gene therapy vector comprising any expression cassette described in invention 1001 to 1021. [Invention 1024] The vector of the present invention 1023, wherein the gene therapy vector is a recombinant adeno-associated virus (rAAV) vector. [Invention 1025] The vector of the present invention 1024, wherein the rAAV vector is an AAV6, AAV8, AAV9, AAVrh.74, or AAVrh.10 vector or a functional variant thereof. [Invention 1026] The rAAV vector is not an AAV2 vector, and is a vector according to Invention 1024 or Invention 1025. [Invention 1027] A vector according to any of the inventions 1024 to 1026, wherein the rAAV vector contains a capsid protein that shares 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with any one of sequence numbers 15 to 17. [Invention 1028] A method for treating and / or preventing a disease or disorder in a subject requiring such treatment, comprising the step of administering a vector according to any of invention 1023 to 1027 to the subject. [Invention 1029] The method of the present invention 1028, wherein the disease or disorder is a neurological disorder. [Invention 1030] The method of the present invention 1028 or 1029, wherein the disease or disorder is glucose transporter 1 deficiency syndrome (GLUT1DS) or Devibo disease. [Invention 1031] The method according to any of items 1028 to 1030 of the present invention, wherein the vector is administered by intraventricular (ICV) injection. [Invention 1032] Any method of the present invention 1028-1031, wherein the administration results in the expression of the polynucleotide sequence encoding GLUT1 in the brain at an arbitrarily increased level compared to a reference rAAV vector. [Invention 1033] The method of any of the invention 1028-1032 wherein the administration results in an increase in the expression of GLUT1 protein in the brain, and / or an increase in glucose and / or lactate levels in CSF, at an optionally increased level compared to a reference rAAV vector, wherein the increase is optionally at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, or more. [Invention 1034] The method according to any one of the present invention 1028 to 1033, wherein the vector is administered in a dose of 1E12 vector genome (vg), 1E13vg, 1E14vg, or 3E14vg. [Invention 1035] Any method of the present invention 1028-1034 that causes increased glucose uptake by cerebral microvascular endothelial cells compared to methods performed using an endogenous Glut1 promoter or a ubiquitous promoter. [Invention 1036] A method for expressing GLUT1 in cells, comprising the step of contacting the cells with any of the vectors of the present invention 1023 to 1027. [Invention 1037] The method of the present invention 1036, wherein the cells are endothelial cells. [Invention 1038] The method of the present invention 1037, wherein the endothelial cells are cerebral microvascular endothelial cells. [Invention 1039] The method of the present invention 1037 or 1038, wherein the endothelial cells are in vivo endothelial cells. [Invention 1040] The method of the present invention 1036, wherein the cells are neurons. [Invention 1041] The method of the present invention 1040, wherein the neuron is an in vivo neuron. [Invention 1042] A method according to any of the present invention 1036 to 1040, comprising in vivo administration of the vector to a subject. [Invention 1043] Any method according to item 1036 to 1041 of the present invention, wherein the vector causes an increase in glucose uptake by the cells compared to cells that have come into contact with a vector containing an endogenous Glut1 promoter or a ubiquitous promoter. [Invention 1044] A pharmaceutical composition comprising any vector according to invention 1023 to 1027. [Invention 1045] A kit comprising any vector according to Invention 1023-1027 or a pharmaceutical composition according to Invention 1043 and optionally instructions for use. Various other aspects and embodiments are disclosed in the detailed description below. The present invention is limited only by the appended claims. [Brief explanation of the drawing]

[0054] [Figure 1-1] Figure 1 shows vector diagrams for various non-restrictive examples of vector genomes. [Figure 1-2] Same as above. [Figure 2-1] Figure 2 shows a vector diagram of a non-restrictive example of a vector genome. The complete polynucleotide sequence of the vector genome is sequence number 17. The capitalized portion is the expression cassette (sequence number 8). [Figure 2-2] Same as above. [Figure 3-1] Figure 3 shows a vector diagram of a non-restrictive example of a vector genome. The complete polynucleotide sequence of the vector genome is SEQ ID NO: 19. The capitalized portion is the expression cassette (SEQ ID NO: 10). [Figure 3-2] Same as above. [Figure 4-1] Figure 4 shows a vector diagram of a non-restrictive example of a vector genome. The complete polynucleotide sequence of the vector genome is SEQ ID NO: 21. The capitalized portion is the expression cassette (SEQ ID NO: 12). [Figure 4-2] Same as above. [Figure 5-1]Figure 5 shows a vector diagram of a non-restrictive example of a vector genome. The complete polynucleotide sequence of the vector genome is SEQ ID NO: 96. The capitalized portion is the expression cassette (SEQ ID NO: 97). A substitute for the complete polynucleotide sequence of the vector genome is SEQ ID NO: 23. A substitute for the expression cassette is SEQ ID NO: 14. [Figure 5-2] Same as above. [Figure 6-1] Figure 6 shows a vector diagram of a non-restrictive example of a vector genome. The complete polynucleotide sequence of the vector genome is SEQ ID NO: 25. The capitalized portion is the expression cassette (SEQ ID NO: 16). [Figure 6-2] Same as above. [Figure 7-1] Figure 7 shows a vector diagram of a non-restrictive example of a vector genome. The complete polynucleotide sequence of the vector genome is sequence number 98. The capitalized portion is the expression cassette (sequence number 99). [Figure 7-2] Same as above. [Figure 8-1] Figure 8 shows a vector diagram of a non-restrictive example of a vector genome. The complete polynucleotide sequence of the vector genome is sequence number 100. The capitalized portion is the expression cassette (sequence number 101). [Figure 8-2] Same as above. [Figure 9] Figure 9. AAV9-mediated expression of the hGlut1 protein in CHO-Lec2 cells. CHO-Lec2 cells were transduced with an AAV9 vector expressing the hGlut1 transgene protein driven by one of several endothelial-specific promoters (i.e., hFLT1, mTie1, or hGlut1) or the ubiquitous CMV promoter. [SLC2A1=GLUT1 gene] [Figure 10]Figures 10A-10C. Expression of the transgene protein (Glut1-GFP) after transfection in human brain microvascular endothelial cells (hCMEC / d3). Figure 10A. GFP fluorescence 72 hours after transfection with a construct containing one of several endothelial cell promoters that drive the expression of the Glut1-GFP transgene. Figure 10B. GFP fluorescence 72 hours after transfection with a construct containing one of two ubiquitous promoters (CMV or CAG), a control vector without Glut1 (CMV-GFP), or no transfection (no NFX). Images acquired using Operetta CLS® (PerkinElmer®). Figure 10C. Figure of the expression cassette containing the target promoter (hFLT1, mTie, hTie, or hGlut1), and the GLUT1 (SLC2A1) gene (T2A link-GFP), as well as regulatory elements adjacent to the AAV2 inverted terminal repeat (ITR). [Figure 11]Figures 11A-11C. 2-deoxy-D-glucose (glucose) uptake in hCMEC / d3 cells after expression of human GLUT1 (SLC2A1). Human brain microvascular endothelial cells (hCMEC / d3) were transfected with a plasmid expressing either CAG-GFP (negative control), or the hGLUT1-t2A-eGFP transgene driven by one of several endothelial-specific promoters (i.e., hFLT1, mTie1, or hGlut1), or by the ubiquitous CMV promoter. Glucose uptake was measured using a luminescence-based kit (Promega®) with 0.5 mM 2-deoxy-D-glucose (2-DG) in culture medium. Glucose uptake was normalized by total cells using phase-contrast imaging [error bars represent SEM; n=6 replications per state]. Figure 11A. Glucose (2-DG) uptake was measured in the first experiment, 72 hours after transfection. Figure 11B. Glucose (2-DG) uptake was measured 72 hours after transfection in the second experiment. Figure 11C. Glucose (2-DG) uptake was measured 96 hours after transfection. [Figure 12]Figures 12A-12B. 2-deoxy-D-glucose (glucose) uptake in hCMEC / d3 cells after expression of human GLUT1 (SLC2A1). Human brain microvascular endothelial cells (hCMEC / d3) were transfected with a plasmid expressing the hGLUT1-t2A-eGFP transgene, driven by one of several endothelial-specific promoters (i.e., hFLT1, mTie1, or hGlut1) or by the ubiquitous CMV promoter. Untransfected hCMEC / d3 cells were used as control (CON). Glucose uptake was measured using a luminescence-based kit (Promega®) with various concentrations of 2-deoxy-D-glucose in the culture medium (0 mM, 0.1 mM, 0.5 mM, or 1.0 mM). Glucose uptake was normalized at the cellular level by multiplexing using the RealTime-GloMT cell viability assay (Promega®) performed according to the manufacturer's recommendations. Figure 12A shows glucose uptake in hCMEC / d3 cells after human Glut1 (SLC2A1) expression at 72 hours. Figure 12B shows glucose uptake in hCMEC / d3 cells after human Glut1 (SLC2A1) expression at 96 hours. [Figure 13]Figure 13. 2-deoxy-D-glucose (glucose) uptake after AAV9-mediated expression of hGLUT1 (SLC2A1) in hCMEC / d3 cells. Human brain microvascular endothelial cells (hCMEC / d3) were transduced using an AAV9 vector (3x10⁵ vector genome / cell) expressing either CAG-GFP (negative control), or the hGLUT1 transgene driven by one of several endothelial-specific promoters (i.e., hFLT1, mTie1, or hGlut1), or by the ubiquitous CMV promoter. Glucose (2-DG) uptake was measured 72 hours post-transduction using the luminescence-based Glucose Uptake-Glo kit (Promega®) and normalized per cell using the RealTime-Glo MT cell viability assay (Promega®) [error bars represent SEM; n=4 replications per state]. [Modes for carrying out the invention]

[0055] Detailed description of the invention definition Section headings are intended solely for organizational purposes and should not be construed as limiting the subject matter described to any particular aspect or embodiment.

[0056] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the present invention relates. Similar or equivalent methods and materials may be used in the practice of the present invention, but preferred methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are expressly incorporated by reference in their entirety. In case of any conflict, this specification shall prevail, including definitions. Furthermore, the materials, methods, and examples described herein are illustrative and not intended to limit the scope of the invention.

[0057] All publications and patents described herein are incorporated herein by reference in whole, as if each individual publication or patent were specifically and individually indicated to be incorporated by reference. In case of any conflict, this application shall prevail, including any definitions herein. However, any reference to any reference, article, publication, patent, patent publication, and patent application cited herein shall not be, and should not be construed as, an endorsement or proposal in any form of which that constitutes valid prior art or forms part of the common general knowledge in any country of the world.

[0058] In this document, unless otherwise indicated, any concentration range, percentage range, ratio range, or integer range should be understood to include any integer value within the enumerated range, and, where appropriate, fractions thereof (e.g., one-tenth and one-hundredth of an integer). The term “about” when preceding a number or figure means that the number or figure is within a range of plus or minus 10%. Where used herein, the terms “a” and “an” should be understood to refer to “one or more” of the enumerated components, unless otherwise indicated. The use of substitutes (e.g., “or”) should be understood to mean either one, both, or any combination thereof of the substitutes. The term “and / or” should be understood to mean one or both of the choices. Where used herein, the terms “include” and “comprise” are used synonymously.

[0059] As used herein, the terms “identity” and “identical” refer to the percentage of residues that are perfectly identical in the alignment of a “query” sequence and a “target” sequence, such as the alignment generated by the BLAST algorithm, for polypeptide or polynucleotide sequences. Identity is calculated over the entire length of the target sequence unless otherwise specified. Thus, if, when the query sequence is aligned with the target sequence, at least x% (truncated) of residues in the target sequence are aligned to perfectly match the corresponding residues in the query sequence, then the query sequence “shares at least x% identity” with the target sequence. If the target sequence has variable positions (e.g., residues indicated by X), any alignment to any residue in the query sequence is counted as a match.

[0060] As used herein, “AAV vector” or “rAAV vector” refers to a recombinant vector containing one or more target polynucleotides (or transgenes) flanked by an AAV terminal repeat sequence (ITR). Such an AAV vector may be replicated and packaged into infectious viral particles when present in a host cell transfected with a plasmid encoding and expressing the rep and cap gene products. Alternatively, the AAV vector may be packaged into infectious particles using a host cell stably engineered to express the rep and cap genes.

[0061] As used herein, “AAV virion,” “AAV virus particle,” or “AAV vector particle” refers to a virus particle consisting of at least one AAV capsid protein and a capsidized polynucleotide AAV vector. As used herein, if a particle contains heterologous polynucleotides (i.e., polynucleotides other than the wild-type AAV genome, such as a transgene delivered to a mammalian cell), it is usually referred to as an “AAV vector particle” or simply an “AAV vector.” Thus, the production of an AAV vector particle necessarily involves the production of an AAV vector, since the vector is contained within the AAV vector particle.

[0062] As used herein, “promoter” refers to a polynucleotide sequence that can promote the initiation of RNA transcription from a polynucleotide in eukaryotic cells.

[0063] As used herein, “vector genome” refers to a polynucleotide sequence packaged by a vector (e.g., rAAV virion), including contiguous sequences (in the AAV, inverted terminal repeats). The terms “expression cassette” and “polynucleotide cassette” refer to portions of the vector genome between contiguous ITR sequences. “Expression cassette” means that the vector genome contains at least one gene encoding a gene product operably ligated to an element that drives expression (e.g., a promoter).

[0064] As used herein, the terms “requiring patient” or “requiring subject” refer to a patient or subject at risk of or suffering from a disease, disorder, or condition suitable for treatment or improvement with the recombinant gene therapy vectors or gene editing systems disclosed herein. A requiring patient or subject may, for example, be a patient or subject diagnosed with a central nervous system disorder. The subject may have mutations in the SLC2A1 gene, or deletions of all or part of the SLC2A1 gene, or gene regulatory sequences, which cause abnormal expression of the GLUT1 protein. “Subject” and “patient” are used interchangeably herein. Subjects treated by the methods described herein may be neonates, infants, young children, or adults.

[0065] As used herein, the terms “variant” or “functional variant” interchangeably refer to a protein having one or more amino acid substitutions, insertions, or deletions compared to the parent protein, which retains one or more desired activities of the parent protein.

[0066] As used herein, “genetic disruption” means a partial or complete loss of function or abnormal activity in a gene. For example, an object may suffer genetic disruption in the expression or function of the SLC2A1 gene, resulting in reduced expression of the GLUT1 protein or loss or abnormal function of the GLUT1 protein in at least some of the cells of the object (e.g., endothelial cells and / or neurons).

[0067] As used herein, “to treat” means to improve one or more symptoms of a disease or disorder. The term “to prevent” means to delay or interrupt the onset of one or more symptoms of a disease or disorder, or to slow the progression of an SLC2A1-related neurological disorder or disorder, such as GLUT1 deficiency syndrome (GLUT1 DS). GLUT1 protein or polynucleotide

[0068] This disclosure concerns compositions and methods of use related to the glucose transporter 1 (GLUT1) protein. Various mutations in SLC2A1 are known to be associated with GLUT1 DS. Both hereditary and de novo mutations have been observed. In some cases, heterozygous missense mutations are sufficient to cause disease.

[0069] The polypeptide sequence of GLUT1 is as follows: MEPSSKKLTGRLMLAVGGAVLGSLQFGYNTGVINAPQKVIEEFYNQTWVHRYGESILPTTLTTLWSLSVAIFSVGGMIGSFSVGLFVNRFGRRNSMLMMNLLAFVSAVLMGFSKLGKSFEMLI LGRFIIGVYCGLTTGFVPMYVGEVSPTALRGALGTLHQLGIVVGILIAQVFGLDSIMGNKDLWPLLLSIIFIPALLQCIVLPFCPEESPRFLLINRNEENRAKSVLKKLRGTADVTHDLQEMKE ESRQMMREKKVTILELFRSPAYRQPILIAVVLQLSQQLSGINAVFYYSTSIFEKAGVQQPVYATIGSGIVNTAFTVVSLFVVERAGRRTLHLIGLAGMAGCAILMTIALALLEQLPWMSYLSI VAIFGFVAFFEVGPGPIPWFIVAELFSQGPRPAAIAVAGFSNWTSNFIVGMCFQYVEQLCGPYVFIIFTVLLVLFFIFTYFKVPETKGRTFDEIASGFRQGGASQSDKTPEELFHPLGADSQV (Sequence number: 26).

[0070] In some embodiments, the GLUT1 protein comprises a polypeptide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 26).

[0071] In some embodiments, the disclosure provides a recombinant adeno-associated virus (rAAV) virion comprising a capsid and a vector genome, the vector genome comprising a polynucleotide sequence encoding the GLUT1 protein or a functional variant thereof operably ligated to a promoter. In some embodiments, the disclosure provides a recombinant adeno-associated virus (rAAV) virion comprising a capsid and a vector genome, the vector genome comprising a polynucleotide sequence encoding the GLUT1 protein operably ligated to a promoter. The polynucleotide encoding the GLUT1 protein may comprise a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the following: (Sequence ID 5)

[0072] In some embodiments, the polynucleotide sequence encoding the GLUT1 protein is a codon-optimized sequence. The polynucleotide sequence encoding the GLUT1 protein may include a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the following: (Sequence ID 27)

[0073] Optionally, the polynucleotide sequence encoding the vector genome may include, but is not limited to, a Kozak sequence containing GCCACCATGG (SEQ ID NO: 28). The Kozak sequence may overlap with the polynucleotide sequence encoding the GLUT1 protein or a functional variant thereof. For example, the vector genome may include a polynucleotide sequence (Kozak is underlined) that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the following: gccaccATGG (Sequence ID 29)

[0074] In some embodiments, the Kozak sequence is an alternative Kozak sequence comprising or consisting of one of the following: (gcc)gccRccAUGG(sequence number 30) AGNNAUGN; ANNAUGG; ACCAUGG, and GACACCAUGG (Sequence ID 31).

[0075] In some embodiments, the vector genome does not contain a Kozak sequence.

[0076] Vector genome The AAV virions of this disclosure include a vector genome. The vector genome may include an expression cassette (or a polynucleotide cassette for gene editing applications that do not require the expression of a polynucleotide sequence). Any suitable inverted end repeat (ITR) may be used. The ITR may be derived from the same serotype as the capsid, or from a different serotype (for example, an AAV2 ITR may be used).

[0077] In some embodiments, the 5'ITR includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the following: CCTGCAGGCAGCTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCT (Sequence ID 32)

[0078] In some embodiments, the 5'ITR includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the following: GCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCAGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTA (Sequence ID 6)

[0079] In some embodiments, the 5'ITR includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the following: CTGCGCGCTCGCTCGCTCACTGAGGCCGCCCGGGCAAAGCCCGGGCGTCGGGCGACCTTTGGTCGCCCGGCCTCAGTGAGCGAGCGAGCGCGAGAGGGAGTGGCCAACTCCATCACTAGGGGTTCCTTGTAGTTAATGATTAACCCGCCATGCTACTTATCTACGTA (Sequence ID 33)

[0080] In some embodiments, the 3'ITR includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the following: AGGAACCCCTAGTGATGGAGTTGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGCAGCTGCCTGCAGG (Sequence ID 34)

[0081] In some embodiments, the 3'ITR includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the following: TACGTAGATAAGTAGCATGGCGGGTTAATCATTAACTACAAGGAACCCCTAGTGATGGAGTTGCCACTCCCTCTCTGCGCGCTCGCTCGCTCACTGAGGCCGGGCGACCAAAGGTCGCCCGACGCCCGGGCTTTGCCCGGGCGGCCTCAGTGAGCGAGCGAGCGCGC (Sequence ID 7)

[0082] In some embodiments, the vector genome includes one or more filler sequences that are at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to, for example, the following: GCGGCAATTCAGTCGATAACTATAACGGTCCTAAGGTAGCGATTTAAATACGCGCTCTCTTAAGGTAGCCCCGGGACGCGTCAATTGACTACAAACCGAGTATCTGCAGAGGGCCCTGCGTATG (SEQ ID NO: 35) CTTCTGAGGCGGAAAGAACCAGATCCTCTCTTAAGGTAGCATCGAGATTTAAATTAGGGATAACAGGGTAATGGCGCGGGCCGC (SEQ ID NO: 36) GTTACCCAGGCTGGAGTGCAGTGGCACATTTCTGCTCACTGCAACCTCCTCCTCCCTGGGTTC (Sequence No. 37)

[0083] promoter In some embodiments, a polynucleotide sequence encoding the GLUT1 protein or a functional variant thereof is operably ligated to a promoter.

[0084] This disclosure intends to utilize a variety of promoters. Promoters useful in embodiments of this disclosure include, but are not limited to, a cytomegalovirus (CMV) promoter, a phosphoglycerate kinase (PGK) promoter, or a promoter sequence comprising a CMV enhancer, a chicken beta-actin promoter, and a portion of the rabbit beta-globin gene (CAG). In some cases, the promoter may be a synthetic promoter. Examples of synthetic promoters are provided by Schlabach et al PNAS USA 107(6):2538-43 (2010). In some embodiments, the promoter includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the following: ACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACT TGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCT ATTACCATGGTCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCCCATCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTATTTATTTTTTAATTATTTTGTGCAGCGATGGGGGCGGGGGGGGGGGGGCGCGCGCCAGGCG GGGCGGGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCCTTTTATGGCGAGGCGGCGGCGGCGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGGCGG (Sequence No. 38)

[0085] In some embodiments, a polynucleotide sequence encoding the GLUT1 protein, or a functional variant thereof, is operably ligated to an inductive promoter. The inductive promoter may be configured to transcriptionally express or deexpress the polynucleotide sequence in response to the addition or accumulation of a drug, or in response to the removal, degradation, or dilution of a drug. The drug may be a pharmacokinetic. The drug may be, but is not limited to, a tetracycline or one of its derivatives, but includes doxycycline. In some cases, the inductive promoter is a tet-on promoter, a tet-off promoter, a chemically regulated promoter, or a physically regulated promoter (i.e., a promoter that responds to the presence or absence of light, or to low or high temperatures). Inductive promoters include heavy metal ion inductive promoters (such as the mouse mammary tumor virus (mMTV) promoter or various growth hormone promoters), and T7 phage-derived promoters that are active in the presence of T7 RNA polymerase. This list of inductive promoters is non-exclusive.

[0086] In some cases, the promoter is a tissue-specific promoter, such as a promoter that can drive expression in neurons rather than non-neuronal cells. In some embodiments, the tissue-specific promoter is a neuron-specific promoter. In some embodiments, the tissue-specific promoter is selected from any variety of neuron-specific promoters, including but not limited to hSYN1 (human synapsin), INA (alpha internexin), NES (nestin), TH (tyrosine hydroxylase), FOXA2 (forkheadbox A2), CaMKII (calmodulin-dependent protein kinase II), and NSE (neuron-specific enolase). In some cases, the promoter is a ubiquitous promoter. A "ubiquitous promoter" refers to a promoter that is not tissue-specific under experimental or clinical conditions. In some cases, the ubiquitous promoter is one of the following: CMV, CAG, UBC, PGK, EF1-alpha, GAPDH, SV40, HBV, chicken beta-actin, and human beta-actin promoters.

[0087] In some embodiments, the promoter sequence is selected from Table 3. In some embodiments, the promoter includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NOs: 1-3 and 39-51.

[0088] (Table 3) TIFF2026123072000002.tif95166TIFF2026123072000003.tif241166TIFF2026123072000004.tif243166TIFF2026123072000005.ti f243166TIFF2026123072000006.tif243166TIFF2026123072000007.tif232166TIFF2026123072000008.tif241166TIFF20261230720 00009.tif243166TIFF2026123072000010.tif241166TIFF2026123072000011.tif243166TIFF2026123072000012.tif243166TIFF202 6123072000013.tif244166TIFF2026123072000014.tif246166TIFF2026123072000015.tif244166TIFF2026123072000016.tif240166

[0089] In a preferred embodiment, the vector genome includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 1. In a preferred embodiment, the vector genome includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 2. In a preferred embodiment, the vector genome includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO: 3.

[0090] Further exemplary examples of promoters include the SV40 late promoter derived from Simian virus 40, baculovirus polyhedral enhancer / promoter elements, herpes simplex virus thymidine kinase (HSV tk), early promoters derived from cytomegalovirus (CMV), and various retroviral promoters including LTR elements. A wide variety of other promoters are known and commonly available in the art, and sequences of many such promoters are available in sequence databases such as the GenBank database.

[0091] Other adjustment elements In some cases, the vectors of the present disclosure further include one or more regulatory elements selected from the group consisting of enhancers, introns, polyA signals, 2A peptide coding sequences, WPREs (woodchuck hepatitis virus post-transcriptional regulatory elements), and HPREs (hepatitis B post-transcriptional regulatory elements).

[0092] In some embodiments, the vector includes a CMV enhancer.

[0093] In certain embodiments, the vector includes one or more enhancers. In certain embodiments, the enhancers are CMV enhancer sequences, GAPDH enhancer sequences, β-actin enhancer sequences, or EF1-α enhancer sequences. The aforementioned sequences are known in the art. For example, the sequence of the CMV earliest (IE) enhancer is as follows: CGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATGGGACTTTCCATTGACGTCATGGGTGGAGTATTTACGGTAAACTGCCCACT TGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATTAGTCATCGCTATTACCATG (Sequence ID 52)

[0094] In certain embodiments, the vector includes one or more introns. In certain embodiments, the introns are rabbit globin intron sequences, chicken β-actin intron sequences, synthetic intron sequences, or EF1-α intron sequences.

[0095] In certain embodiments, the vector includes a polyA sequence. In certain embodiments, the polyA sequence is a rabbit globin polyA sequence, a human growth hormone polyA sequence, a bovine growth hormone polyA sequence, a PGK polyA sequence, an SV40 polyA sequence, or a TK polyA sequence. In some embodiments, the polyA signal may be a bovine growth hormone polyadenylation signal (bGHpA).

[0096] In certain embodiments, the vector includes one or more transcript stabilizing elements. In certain embodiments, the transcript stabilizing elements are a WPRE sequence, an HPRE sequence, a scaffold attachment region, a 3'UTR, or a 5'UTR. In certain embodiments, the vector includes both a 5'UTR and a 3'UTR.

[0097] In some embodiments, the vector includes a 5' untranslated region (UTR) selected from Table 4. In some embodiments, the vector genome includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one of sequence numbers 53–61.

[0098] (Table 4) TIFF2026123072000017.tif90166TIFF2026123072000018.tif246166TIFF20261230720 00019.tif246166TIFF2026123072000020.tif246166TIFF2026123072000021.tif144166

[0099] In some embodiments, the vector includes a 3' untranslated region selected from Table 5. In some embodiments, the vector genome includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one of sequence numbers 62–70.

[0100] (Table 5) TIFF2026123072000022.tif56166TIFF2026123072000023.tif244166TIFF20261230720 00024.tif246166TIFF2026123072000025.tif242166TIFF2026123072000026.tif41166

[0101] In some embodiments, the vector includes a polyadenylation (poly-A) signal selected from Table 6. In some embodiments, the poly-A signal includes a polynucleotide sequence that is at least 75%, 80%, 85%, 90%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to one of sequence numbers 71-75.

[0102] (Table 6) TIFF2026123072000027.tif148166TIFF2026123072000028.tif141166

[0103] Exemplary vector genomes are shown in Figures 2–8, provided as SEQ ID NOs: 17–25. The capitalized portion of each sequence is an expression cassette (SEQ ID NOs: 8–16, SEQ ID NOs: 97, SEQ ID NOs: 99, and SEQ ID NOs: 101). In some embodiments, the vector genome contains, is essentially, or consists of a polynucleotide sequence that shares at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with one of SEQ ID NOs: 8–16, SEQ ID NOs: 97, SEQ ID NOs: 99, and SEQ ID NOs: 101, which optionally have or do not have an ITR sequence in lowercase. Coding sequences are underlined. Expression cassettes are capitalized.

[0104] Adeno-associated virus vector Adeno-associated virus (AAV) is a replication-deficient parvovirus with a single-stranded DNA genome approximately 4.7 kb long, containing two approximately 145-nucleotide inverted terminal repeats (ITRs). AAV has several known variants, sometimes called serotypes, which are classified by their antigenic epitopes. The nucleotide sequences of the genomes of AAV serotypes are known. For example, the whole genome of AAV-1 is available under GenBank registry number NC_002077, the whole genome of AAV-2 is available under GenBank registry number NC_001401 and Srivastava et al., J. Virol., 45:555-564 (1983), the whole genome of AAV-3 is available under GenBank registry number NC_1829, the whole genome of AAV-4 is available under GenBank registry number NC_001829, the genome of AAV-5 is available under GenBank registry number AF085716, the whole genome of AAV-6 is available under GenBank registry number NC_001862, at least portions of the genomes of AAV-7 and AAV-8 are available under GenBank registry numbers AX753246 and AX753249, respectively, and the genome of AAV-9 is available under GenBank registry number Gao et al. The genome for AAV-10 is provided in al., J. Virol., 78:6381-6388 (2004), the genome for AAV-10 is provided in Mol. Ther., 13(1):67-76 (2006), and the genome for AAV-11 is provided in Virology, 330(2):375-383 (2004). The sequence for the AAVrh.74 genome is provided in U.S. Patent No. 9,434,928, incorporated herein by reference. The cis-acting sequences that lead to viral DNA replication (rep), capsid formation / packaging, and host cell chromosome integration are contained within the AAV ITR. Three AAV promoters (named p5, p19, and p40 from their relative map locations) drive the expression of two AAV internal open reading frames encoding the rep and cap genes.Two rep promoters (p5 and p19), linked to differential splicing of a single AAV intron (nucleotides 2107 and 2227), produce four rep proteins (rep78, rep68, rep52, and rep40) from the rep gene. The rep proteins possess multiple enzymatic properties that ultimately carry out viral genome replication. The cap gene is expressed from the p40 promoter and encodes three capsid proteins: VP1, VP2, and VP3. Alternative splicing and non-consensus translation initiation sites are involved in the production of the three related capsid proteins. A single consensus polyadenylation site is located at map position 95 of the AAV genome. The life cycle and genetics of AAV are outlined in Muzyczka, Current Topics in Microbiology and Immunology, 158:97-129 (1992).

[0105] AAV possesses unique characteristics that make it attractive as a vector for delivering foreign DNA to cells, for example, in gene therapy. AAV infection of cells in culture is non-cellular, and natural infection in humans and other animals is asymptomatic. Furthermore, AAV infects many mammalian cells and allows for the potential to target many different tissues in vivo. Additionally, AAV transduces into slow-dividing and non-dividing cells and can essentially persist for the lifetime of these cells as a transcriptionally active nuclear episome (extrachromosomal element). The AAV proviral genome is inserted as cloned DNA within a plasmid, enabling the construction of a recombinant genome. Furthermore, since the signals directing AAV replication and genome capsidation are contained within the ITR of the AAV genome, some or all of the approximately 4.3kb inside the genome (encoding the replication and structural capsid protein, rep-cap) may be replaced with foreign DNA. To generate an AAV vector, the rep and cap proteins may be provided in trans. Another important characteristic of AAV is that it is a very stable and potent virus. AAV readily tolerates the conditions used to inactivate adenoviruses (56°C to 65°C for several hours), reducing the importance of low-temperature storage of AAV. AAV can also be freeze-dried. Finally, AAV-infected cells are not resistant to co-infection.

[0106] The AAV DNA in the rAAV genome may be derived from any AAV variant or serotype from which recombinant virus can be induced, including, but not limited to, AAV-1, AAV-2, AAV-3, AAV-4, AAV-5, AAV-6, AAV-7, AAV-8, AAV-9, AAV-10, AAV-11, AAV-12, AAV-13, and AAVrh10. The production of pseudotyped rAAV is disclosed, for example, in WO01 / 83692. Other types of rAAV variants, such as rAAV with capsid mutations, are also intended. See, for example, Marsic et al., Molecular Therapy, 22(11):1900-1909 (2014). Nucleotide sequences of genomes of various AAV serotypes are publicly known in the art.

[0107] In some cases, rAAVs contain a self-complementary genome. As defined herein, rAAVs containing a “self-complementary” or “double-stranded” genome refer to rAAVs that have been engineered so that the coding region of the rAAV forms an intramolecular double-stranded DNA template, as described by McCarty et al. Self-complementary recombinant adeno-associated virus (scAAV) vectors facilitate efficient transduction independent of DNA synthesis. Gene Therapy. 8(16):1248-54(2001). This disclosure intends to use rAAVs containing a self-complementary genome in some cases, so that, at the time of infection (such as transduction), the two complementary halves of the scAAV immediately form a single double-stranded DNA (dsDNA) unit that can replicate and transcribe, rather than waiting for cell-mediated synthesis of the second strand of the rAAV genome. It should be understood that, instead of the full coding capacity (4.7-6kb) seen in rAAVs, rAAVs containing a self-complementary genome can only hold about half that amount (about 2.4kb).

[0108] In other cases, rAAV vectors contain a single-stranded genome. As defined herein, a “single standard” genome refers to a genome that is not self-complementary. In most cases, non-recombinant AAVs have a single-stranded DNA genome. There have been some indications that rAAVs should be scAAVs to achieve efficient transduction of cells. However, this disclosure intends for rAAV vectors that may have a single-stranded genome rather than a self-complementary genome, based on the understanding that other genetic modifications of rAAV vectors may be beneficial in obtaining optimal gene transcription in target cells. In some cases, this disclosure relates to a single-stranded rAAV vector that can achieve efficient gene transfer to the anterior segment of the mouse eye. See Wang et al. Single stranded adeno-associated virus achieves efficient gene transfer to anterior segment in the mouse eye. PLoS ONE 12(8):e0182473(2017).

[0109] In some cases, the rAAV vector is a vector for serotype AAV1, AAV2, AAV4, AAV5, AAV6, AAV7, AAV8, AAV9, AAV10, AAV11, AAV12, AAV13, AAVrh10, or AAVrh74. The production of pseudotyped rAAV is disclosed, for example, in WO 01 / 83692. Other types of rAAV variants, such as rAAV with capsid mutations, are also intended. See, for example, Marsic et al., Molecular Therapy, 22(11):1900-1909 (2014). In some cases, the rAAV vector is a vector for serotype AAV9. In some embodiments, the rAAV vector is a vector for serotype AAV9 and contains a single-stranded genome. In some embodiments, the rAAV vector is a vector for serotype AAV9 and contains a self-complementary genome. In some embodiments, the rAAV vector contains a sequence of inverted terminal repeats (ITRs) of AAV2. In some embodiments, the rAAV vector contains the AAV2 genome, and as a result, the rAAV vector is an AAV-2 / 9 vector, an AAV-2 / 6 vector, or an AAV-2 / 8 vector.

[0110] The full-length sequence and capsid gene sequence for the most well-known AAV are provided in U.S. Patent No. 8,524,446, which is incorporated herein by reference in its entirety.

[0111] The AAV vector may contain a wild-type AAV sequence or one or more modifications to the wild-type AAV sequence. In certain embodiments, the AAV vector contains one or more amino acid modifications, such as substitutions, deletions, or insertions, within the capsid protein, e.g., VP1, VP2, and / or VP3. In certain embodiments, the modifications result in reduced immunogenicity when the AAV vector is provided to a subject.

[0112] The rAAV capsid protein may be modified so that rAAV targets specific target tissues, such as endothelial cells or, more specifically, endothelial apical cells. In some embodiments, rAAV is injected directly into the target cerebral ventricles.

[0113] In some embodiments, the rAAV virion is an AAV2 rAAV virion. Many of the capsids are AAV2 capsids or functional variants thereof. In some embodiments, the AAV2 capsid shares at least 98%, 99%, or 100% identity with reference AAV2 capsids such as the following: MAADGYLPDWLEDTLSEGIRQWWKLKPGPPPPKPAERHKDDSRGLVLPGYKYLGPFNGLDKGEPVNEADAAALEHDKAYDRQLDSGDNPYLKYNHADAEFQERLKEDTSFGGNLGRAVFQAKKRVLEPLGLVEEPVKTAPGKKRPVEHSPVEPDSSSGTGKAGQQPARKRLNFGQTGDADSVP DPQPLGQPPAAPSGLGTNTMATGSGAPMADNNEGADGVGNSSGNWHCDSTWMGDRVITTSTRTWALPTYNNHLYKQISSQSGASNDNHYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNDGTTTIANNLTSTVQVFTDSEYQLPYVLGSAHQGCLPPFPA DVFMVPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTNTPSGTTTQSRLQFSQAGASDIRDQSRNWLPGPCYRQQRVSKTSADNNNSEYSWTGATKYHLNGRDSLVNPGPAMASHKDDEEKFFPQSGVLIFGKQGSEKTN VDIEKVMITDEEEIRTTNPVATEQYGSVSTNLQRGNRQAATADVNTQGVLPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPPPQILIKNTPVPANPSTTFSAAKFASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYNKSVNVDFTVDTNGVYSEPRPIGTRYLTRNL (Sequence ID 76)

[0114] In some embodiments, the rAAV virion is an AAV9 rAAV virion. Many of the capsids are AAV9 capsids or functional variants thereof. In some embodiments, the AAV9 capsids share at least 98%, 99%, or 100% identity with reference AAV9 capsids such as the following: MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPGNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGNGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTESVPD PQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFP ADVFMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSKTINGSGQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAWPGASSWALNGRNNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDN VDADKVMITNEEEIKTTNPVATESYGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYSEPRPIGTRYLTRNL (Sequence ID 77)

[0115] In some embodiments, the rAAV virion is an AAV6 rAAV virion. Many of the capsids are AAV6 capsids or functional variants thereof. In some embodiments, the AAV6 capsid shares at least 98%, 99%, or 100% identity with reference AAV6 capsids such as the following: MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDDGRGLVLPGYKYLGPFNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLRYNHADAEFQERLQEDTSFGNGNLGRAVFQAKKRVLEPFGLVEEGAKTAPGKKRPVEQSPQEPDSSSGIGKTGQQPAKKRLNFGQTGDSESVPD PQPLGEPPATPAAVGPTTMASGGGAPMADNNEGADGVGNASGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQISSASTGASNDNHYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTTNDGVTTIANNLTSTVQVFSDSEYQLPYVLGSAHQGCLPPFPA DVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFTFSYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLNRTQNQSGSAQNKDLLFSRGSPAGMSVQPKNWLPGPCYRQQRVSKTKTDNNNSNFTWTGASKYNLNGRESIINPGTAMASHKDDKDKFFPMSGVMIFGKESAGASN TALDNVMITDEEEIKATNPVATERFGTVAVNLQSSSTDPATGDVHVMGALPGMVWQDRDVYLQGPIWAKIPHTDGHFHPSPLMGGFGLKHPPPQILIKNTPVPANPPAEFSATKFASFITQYSTGQVSVEIEWELQKENSKRWNPEVQYTSNYAKSANVDFTVDNNGLYTEPRPIGTRYLTRPL (Sequence ID 78)

[0116] In some embodiments, the rAAV virion is an AAVrh.10 rAAV virion. Many of the capsids are AAVrh.10 capsids or functional variants thereof. In some embodiments, the AAVrh.10 capsid shares at least 98%, 99%, or 100% identity with reference AAVrh.10 capsids such as the following. MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDDGRGLVLPGYKYLGPFNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLRYNHADAEFQERLQEDTSFGNGNLGRAVFQAKKRVLEPLGLVEEGAKTAPGKKRPVEPSPQRSPDSSTGIGKKGQQPAKKRLNFGQTGDSESVP DPQPIGEPPAGPSGLGSGTMAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQISNGTSGGSTNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPFP ADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFEFSYQFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQSTGGTAGTQQLLFSQAGPNNMSAQAKNWLPGPCYRQQRVSTTLSQNNNSNFAWTGATKYHLNGRDSLVNPGVAMATHKDDEERFFPSSGVLMFGKQGAGKD NVDYSSVMLTSEEEIKTTNPVATEQYGVVADNLQQQNAAPIVGAVNSQGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPADPPTTFSQAKLASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTDGTYSEPRPIGTRYLTRNL (Sequence ID 79)

[0117] In some embodiments, the rAAV virion is an AAV8 rAAV virion. Many of the capsids are AAV8 capsids or functional variants thereof. In some embodiments, the AAV8 capsid shares at least 98%, 99%, or 100% identity with reference AAV8 capsids such as the following: MAADGYLPDWLEDNLSEGIREWWALKPGAPKPKANQQKQDDGRGLVLPGYKYLGPFNGLDKGEPVNAADAAALEHDKAYDQQLQAGDNPYLRYNHADAEFQERLQEDTSFGGNLGRAVFQAKKRVLEPLGLVEEGAKTAPGKKRPVEPSPQRSPDSSTGIGKKGQQPARKRLNFGQTGDSESVP DPQPLGEPPAAPSGVGPNTMAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQISNGTSGGATNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLSFKLFNIQVKEVTQNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPPFP ADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFQFTYTFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQTTGGTANTQTLGFSQGGPNTMANQAKNWLPGPCYRQQRVSTTTGQNNNSNFAWTAGTKYHLNGRNSLANPGIAMATHKDDEERFFPSNGILIFGKQNAARD NADYSDVMLTSEEEIKTTNPVATEEYGIVADNLQQQQNTAPQIGTVNSQGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPADPPTTFNQSKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTSVDFAVNTEGVYSEPRPIGTRYLTRNL (Sequence ID 80)

[0118] In some embodiments, the rAAV virion is an AAVrh.74 rAAV virion. Many of the capsids are AAVrh.74 capsids or functional variants thereof. In some embodiments, the AAVrh.74 capsid shares at least 98%, 99%, or 100% identity with reference AAVrh.74 capsids such as the following. MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDNGRGLVLPGYKYLGPFNGLDKGEPVNAADAAALEHDKAYDQQLQAGDNPYLRYNHADAEFQERLQEDTSFGGNLGRAVFQAKKRVLEPLGLVESPVKTAPGKKRPVEPSPQRSPDSSTGIGKKGQQPAKKRLNFGQTGDSESVP DPQPIGEPPAGPSGLGSGTMAAGGGAPMADNNEGADGVGSSSGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQISNGTSGGSTNDNTYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTQNEGTKTIANNLTSTIQVFTDSEYQLPYVLGSAHQGCLPFP ADVFMIPQYGYLTLNNGSQAVGRSSFYCLEYFPSQMLRTGNNFEFSYNFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTQSTGGTAGTQQLLFSQAGPNNMSAQAKNWLPGPCYRQQRVSTTLSQNNNSNFAWTGATKYHLNGRDSLVNPGVAMATHKDDEERFFPSSGVLMFGKQGAGKD NVDYSSVMLTSEEEIKTTNPVATEQYGVVADNLQQQNAAPIVGAVNSQGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGLKHPPPQILIKNTPVPADPPTTFNQAKLASFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSTNVDFAVNTEGTYSEPRPIGTRYLTRNL (Sequence No. 81)

[0119] In some embodiments, the rAAV virion is the AAV-PHP.B rAAV virion or its neurotrophic variant, such as those disclosed in International Patent Publications WO2015 / 038958A1 and WO2017 / 100671A1, but is not limited thereto. For example, the AAV capsid may comprise at least four consecutive amino acids from the sequence TLAVPFK (SEQ ID NO: 83) or KFPVALT (SEQ ID NO: 84), which are inserted, for example, between the sequences encoding amino acids 588 and 589 of AAV9.

[0120] Many capsids are AAV-PHP.B capsids or functional variants thereof. In some embodiments, AAV-PHP.B capsids share at least 98%, 99%, or 100% identity with reference AAV-PHP.B capsids such as the following: MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGLVLPGYKYLGPGNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERLKEDTSFGNGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSGAQPAKKRLNFGQTGDTESVPDP QPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVGSSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPWGYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTSTVQVFTDSDYQLPYVLGSAHEGCLPPFPADV FMIPQYGYLTLNDGSQAVGRSSFYCLEYFPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTINGSGQNQQTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAWPGASSWALNGRNNSLMNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVDADK VMITNEEEIKTTNPVATESYGQVATNHQSAQTLAVPFKAQAQTGWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYSEPRPIGTRYLTRNL (Sequence No. 82)

[0121] Further AAV capsids used in the rAAV vilion of this disclosure include those disclosed in International Patent Publications WO2009 / 012176A2 and WO2015 / 168666A2.

[0122] While not bound by theory, the inventors determined that the AAV9 vector or the AAVrh.10 vector would confer broad CNS distribution to the vector. Also, while not bound by theory, the inventors further determined that the AAV6 vector may provide some specificity to target endothelial cells. Other vector serotypes, including but not limited to AAV8 and AAVrh.10, may be used.

[0123] In some embodiments, the rAAV vector is not an AAV2 vector. Not bound by theory, the inventors determined that, in some cases, the use of an AAV2 vector results in the transduction of neurons in addition to, or instead of, endothelial cells. Not bound by theory, the inventors further determined that the spread of an AAV2 vector within the CNS is limited by its interaction with heparan sulfate proteoglycan (HSPG) receptors.

[0124] Pharmaceutical compositions and kits In one embodiment, the Disclosure provides a pharmaceutical composition comprising the rAAV virion of the Disclosure and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0125] For example, various solutions, such as sterile aqueous solutions, can be used for administration by injection. Such aqueous solutions may be buffered if desired, and the liquid diluent can be initially isotonic with physiological saline or glucose. Solutions of rAAV as a free acid (DNA contains acidic phosphate groups) or a pharmacokinetically acceptable salt can be prepared in water preferably mixed with a surfactant such as 0.001% or 0.01% Poloxamer 188. Dispersions of rAAV may be prepared in glycerol, liquid polyethylene glycol and mixtures thereof, and oil. Under normal storage and use conditions, these preparations contain preservatives to prevent microbial growth. In this regard, all sterile aqueous media used are readily available by standard techniques well known to those skilled in the art.

[0126] Suitable pharmaceutical forms for injectable use include, but are not limited to, sterile aqueous solutions or dispersions, and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and fluid enough to be readily injectable (syringability). It must be stable under manufacturing and storage conditions and protected against microbial contamination, such as bacteria and fungi. The carrier may be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oils. Adequate fluidity can be maintained, for example, by the use of coatings such as lecithin, by maintaining the required particle size in the case of dispersions, and by the use of surfactants. Prevention of microbial action can be provided by various antimicrobial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. It is often preferable to include isotonic agents, such as sugars or sodium chloride. Long-term absorption of injectable compositions can be achieved by using absorption-delaying agents, such as aluminum monostearate and gelatin.

[0127] Sterile injectable solutions can be prepared by incorporating the required amount of rAAV in a suitable solvent, along with various other components listed above as needed, and then sterilizing by filtration. Generally, dispersants are prepared by incorporating the sterile active ingredient into a sterile vehicle containing a basic dispersion medium and other components required from those listed above. For sterile powders for the preparation of sterile injectable solutions, preferred preparation methods are vacuum drying and freeze-drying techniques, which yield powders of the active ingredient plus any additional desired components from a pre-sterilized filtered solution.

[0128] In another embodiment, the Disclosure includes a kit comprising the rAAV virion and instructions for use.

[0129] How to use In one embodiment, the Disclosure provides a method for increasing GLUT1 activity in cells, the method comprising contacting cells with rAAV of the Disclosure. In another embodiment, the Disclosure provides a method for increasing GLUT1 activity in a subject, the method comprising administering rAAV of the Disclosure. In some embodiments, cells and / or subjects are deficient in the expression level and / or activity of SLC2A1 messenger RNA or GLUT1 protein and / or include loss-of-function mutations in SLC2A1. The cells may be endothelial cells, for example, endothelial apical cells.

[0130] In some embodiments, the method restores normal function of endothelial apical cells. In some embodiments, the method restores GLUT1 transporter protein expression levels in cell culture and / or in vivo. In some embodiments, the method restores normal glucose transport and metabolism (e.g., glycolysis, lactate production) in cell culture and / or in vivo. In some embodiments, the method restores normal angiogenesis and / or microvascular development in the central nervous system (CNS).

[0131] Treatment method In another embodiment, the Disclosure provides a method for treating a disease or disorder in a subject requiring treatment, the method comprising administering an effective amount of the rAAV virion of the Disclosure to the subject. In some embodiments, the disease or disorder is a neurological disorder or disorder. In some embodiments, the subject suffers from a genetic disruption in SLC2A1 expression or function. In some embodiments, the disease or disorder is GLUT1 deficiency syndrome (GLUT1 DS).

[0132] AAV-mediated delivery of the GLUT1 protein to the CNS may extend lifespan and prevent, reduce, mitigate, or diminish neurodegeneration, early-onset seizures, developmental delay, acquired microcephaly (slowed head growth), complex motor impairments (spasticity, ataxia, dystonia), paroxysmal intraocular movements, and / or low lactate and / or low cerebrospinal fluid glucose levels (CSF hypoglycemia). In some embodiments, this method provides, for example, early treatment of the disease course in neonates, infants, or young children.

[0133] The methods disclosed herein may provide efficient biodistribution in the brain and / or CNS. They may result in sustained expression in all or a significant proportion of endothelial cells (e.g., endothelial apical cells). In particular, the methods disclosed herein may provide long-term, sustained expression of the GLUT1 protein throughout the growth and aging of the subject.

[0134] Combination therapy is also envisioned by the present invention. Combinations of the methods of the present invention with standard medical treatments (e.g., corticosteroids or topical decompressants) are specifically envisioned, as are combinations with novel therapies. In some cases, the subject may be treated with a combination of steroids and / or immunosuppressants to prevent or reduce the immune response to the administration of rAAV as described herein.

[0135] A therapeutically effective dose of rAAV vector, for example, intraventricular (ICV) or intracisional (ICM) injection, is a dose of rAAV of approximately 1 e12 vg / kg to approximately 5 e12 vg / kg, or approximately 1 e13 vg / kg to approximately 5 e13 vg / kg, or approximately 1 e14 vg / kg to approximately 5 e14 vg / kg, or approximately 1 e15 vg / kg to approximately 5 e15 vg / kg in terms of brain weight. The present invention also includes compositions comprising rAAV vectors within these ranges.

[0136] For example, in certain embodiments, therapeutically effective doses of rAAV vector are doses of about 1 e10 vg, about 2 e10 vg, about 3 e10 vg, about 4 e10 vg, about 5 e10 vg, about 6 e10 vg, about 7 e10 vg, about 8 e10 vg, about 9 e10 vg, about 1 e12 vg, about 2 e12 vg, about 3 e12 vg, about 4 e12 vg, about 4 e13 vg, and 4 e14 vg. The present invention also includes compositions comprising these doses of rAAV vector.

[0137] In some embodiments, for example, when ICV injection is performed, the therapeutically effective dose of rAAV vector is in the range of 1e10vg / hemisphere to 2e14vg / hemisphere, or approximately 1e10vg / hemisphere, approximately 1e11vg / hemisphere, approximately 1e12vg / hemisphere, approximately 1e13vg / hemisphere, or approximately 1e14vg / hemisphere. In some embodiments, for example, when ICM injection is performed, the therapeutically effective dose of rAAV vector is in the range of 2e10vg to 2e14vg total, or approximately 2e10vg total, approximately 2e11vg total, approximately 2e12vg total, approximately 2e13vg total, or approximately 2e14vg total.

[0138] In some embodiments, the therapeutic composition contains more than approximately 1e9, 1e10, or 1e11 of rAAV vector genomes per volume of the injected therapeutic composition. In some embodiments, the therapeutic composition contains more than approximately 1e11, 1e12, 1e13, or 1e14 of rAAV vector genomes per mL. In certain embodiments, the therapeutic composition contains less than approximately 1e14, 1e13, or 1e12 of rAAV vector genomes per mL.

[0139] Evidence of functional improvement, clinical utility, or effectiveness in patients may be assessed by analyzing paroxysmal intraocular movements, surrogate markers of seizure frequency reduction (generalized tonic-clonic seizures and myoclonic seizures), cerebrospinal fluid lactate and / or glucose concentrations (CSF), and evaluation of developmental disorders, chorea, dystonia, and microcephaly. Standard disease assessment scales such as the Columbia Neurological Score, Composite Intellectual Estimate, Adaptive Behavior Composite, verbal and nonverbal cognitive abilities, visual-motor integration, and the 6-minute walk test are used to measure cognitive, motor, and language functions. Cognitive and developmental assessments, including the Peabody Developmental Motor Scale II (PDMS-2) and the Bailey Infant Development Scale III, are applied according to the child's level of disability. Gross motor function measurement (GFMF-88) and Pediatric Disability Inventory (PEDI) are also used. These scales, or similar scales, as well as patient-reported QOL outcomes, such as the three-point scale (decreased mean duration, no change, increase) Caregiver Global Impression of Change in Seizure Duration (CGICSD), Pediatric Quality of Life Inventory (PedsQL®), and Vineland Adaptive Behavior Scales-2nd, may demonstrate improvement in disease components. Baseline and post-treatment magnetic resonance imaging may show age-appropriate improvement or normalization of brain volume compared to age-adjusted control data and historical data from GLUT1 deficiency patients.

[0140] Clinical benefits can be confirmed by extension of lifespan, normalization of neurodevelopment, normalization of cerebrospinal fluid glucose concentration, reduction in the frequency or magnitude of paroxysmal intraocular movements, reduction or elimination of epileptic seizure activity (such as myoclonic, clonic, generalized rigid-clonic, and / or epileptic seizures), improvement or resolution of complex motor disorders such as spasticity, dystonia, and / or ataxia, and improvement or normalization of the Columbia Neurological Score and / or 6-minute walk test. Evidence of neuroprotective and / or neuroregenerative effects may be evident by characterizing overall brain size, microcephaly, and / or cortical and / or cerebellar atrophy in all the aforementioned metrics and / or magnetic resonance imaging (MRI).

[0141] In some embodiments, the method results in increased glucose uptake by cells compared to cells contacted with a vector containing an endogenous Glut1 promoter or a ubiquitous promoter, or to target cells to which the vector was administered. In some cases, the increase is at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, or at least 50%. In some cases, the increase is at least 1.1 times, at least 1.2 times, at least 1.3 times, at least 1.4 times, at least 1.5 times, at least 1.6 times, at least 1.7 times, or at least 1.8 times. The vector may be any vector disclosed herein. The cells may be endothelial cells or nerve cells. For example, the method may increase glucose uptake by human brain microvascular endothelial cells, either in vitro or in vivo.

[0142] Administration of composition The effective dose of the composition may be administered by standard routes of the art, including, but not limited to, intravenous, intracerebral, intrathecal, intracisional, or intracerebroventricular administration. In some cases, administration may include intravenous, intracerebral, subarachnoid, intracisional, or intracerebroventricular injection. Administration may be performed by subarachnoid injection with or without Trendelenburg tilt. Intracisional (ICM) delivery may be achieved via catheter insertion in the intrathecal (IT) space. Intracerebroventricular injection may be achieved via neurosurgical targeting guided by magnetic resonance imaging (MRI).

[0143] In some embodiments, this disclosure provides a systemic administration of an effective dose of the rAAV and compositions of the present invention. For example, systemic administration may be administered into the circulatory system so that the whole body is affected. Systemic administration includes intravenous administration by injection or infusion.

[0144] In particular, the administration of rAAV according to the present invention can be achieved by using any physical method for delivering the rAAV recombinant vector to the target tissue of an animal. Administration includes, but is not limited to, injection into the central nervous system (CNS) or cerebrospinal fluid (CSF), and / or direct injection into the brain.

[0145] In some embodiments, the methods of the Disclosure include intraventricular, intracisional, subarachnoid, or intraparenchymal delivery. Infusion may be performed using an infusion pump, a dedicated cannula, catheter, or syringe / needle. Optionally, targeting of the injection site may be achieved using MRI-guided imaging. Administration may include delivery of an effective amount of rAAV virion, or a pharmaceutical composition containing rAAV virion, to the CNS. These can be achieved, for example, via unilateral intraventricular infusion, bilateral intraventricular infusion, intracisional magna infusion with Trendelenburg tilt procedure, or intracisional magna infusion without Trendelenburg tilt procedure, subarachnoid infusion with Trendelenburg tilt procedure, or subarachnoid infusion without Trendelenburg tilt procedure. The compositions of the Disclosure may further be administered intravenously.

[0146] Direct delivery to the central nervous system (CNS) may involve targeting the ventricles, unilateral or bilateral, specific neuronal regions, or more general brain regions including neuronal targets. The selection of intraventricular space, brain region, and / or neuronal target for individual patients, as well as subsequent intraoperative delivery of AAV, can be achieved by employing any number of software planning programs (e.g., Stealth System, Clearpoint Neuronavigation System, Brainlab, Neuroinspire, etc.) using a variety of imaging techniques (MRI, CT, CT combined with MRI merging). Targeting and delivery of intraventricular space or brain region may involve the use of standard stereotactic frames (Leksell, CRW) or a frameless approach with or without intraoperative MRI. Actual delivery of AAV may be by injection through a needle or cannula with or without a material-coated inner lumen to prevent adsorption of the AAV vector (e.g., Smartflow cannula, MRI Interventions cannula). The delivery device consists of a syringe and an automated or microinjection pump with pre-programmed injection rate and volume. A syringe / needle combination, or a guide cannula for the needle alone, may be connected to a stereotactic frame via a direct interface. Infusion may involve a constant flow rate or a variable rate due to convective-enhanced delivery. [Examples]

[0147] Example 1: Preclinical biological activity and efficacy Recombinant AAV virions are produced using the vector genomes disclosed in Figures 2–8. These are evaluated in disease model mice as a result of GLUT1 deficiency disease. One model involves floxing the GLUT1 gene and crossing it with genetically modified animals expressing Cre / lox from a constitutive promoter or an endothelial-specific promoter (e.g., Tie-2). The resulting mice are heterozygous nulls at the GLUT1 locus and exhibit developmental phenotypes that mimic human disease. A second model mouse for GLUT1 deficiency is a heterozygous haploinsufficient mouse (GLUT-1) created by targeted disruption of the promoter and exon 1 region of the mouse GLUT-1 gene. + / - This is the mouse model. Other animal models may include the GLUT1 DS model, in which the GLUT1 gene has a S324P point mutation.

[0148] Gene expression and dose-response are evaluated in vitro (using endothelial and neuronal cell lines) and in vivo (using wild-type and GLUT1 DS model mice). Cultured cells (human embryonic kidney cells 293, HEK293; human umbilical vein endothelial cells, HUVEC; human brain-derived endothelial cells, bEND3; human brain microvascular endothelial cells, HBEC-5i; human brain microvascular endothelial cell line, hCMEC / D3 (blood-brain barrier model); human glial oligodendrosis hybrid cells, MO3.13; human neuroblastoma, SH-SY5Y transfected with SLC2A1 expression vector) are analyzed for transduction efficiency by quantitative real-time PCR and GLUT1 levels by ELISA and / or Western blotting. The concept and efficacy of the AAV vector construct will be demonstrated in vivo using GLUT1 DS mice, compared to GLUT1 DS mutant control mice, by immunolabeling of transgene (GLUT1 protein) expression in the CNS, increased cerebral capillary density and / or increased vascular size in the CNS, increased cerebral glycation by positron emission tomography (PET), increased CSF glucose or lactate levels and / or CSF / blood glucose ratio, increased CSF lactate levels, and improved motor performance as measured by standard assessments such as rotational load and vertical pole evaluation. In vivo gene expression and efficacy using GLUT1 DS mice will be demonstrated by administering the AAV vector construct intravenously or directly into the ventricle, using these administration routes alone or in combination.

[0149] Example 2: In vitro evaluation of GLUT1 expression using an endothelial promoter Gene expression was evaluated in vitro using human brain microvascular endothelial cells (hCMEC / D3). Glut1 expression was assessed in hCMEC / D3 cells transfected with an AAV9 vector encoding SLC2A1 under the control of the hFLT1, mTIE1, hGlut1, or CMV promoter (illustrated in Figure 10C) (Figure 9). Expression from the endothelial promoters (hFLT1 and mTIE1) was comparable to expression from the Glut1 promoter and significantly lower than expression from the CMV promoter. Similar patterns of expression levels among these constructs were observed by immunofluorescence microscopy (Figures 10A and 10B).

[0150] Surprisingly, 2-deoxy-D-glucose (2-DG) uptake by human brain microvascular endothelial cells transfected or transduced under the control of the endothelial promoter was greater than that by the regulatory Glut1 promoter, with the hFLT-1 promoter showing the highest levels of 2-DG (glucose) uptake (Figures 11A–11C, 12, and 13). The discovery of greater 2-DG (glucose) uptake in the hFLT-1 promoter construct was also observed across a range of 2-DG concentrations (Figure 12A; 0, 0.1, 0.5, and 1 mM) and at various time points after transfection (Figure 12B), and in some cases was found to be comparable to or slightly greater than that observed with the CMV promoter (Figures 11A–11C; Figures 12A, 12B; Figure 13).

[0151] Figure 9 shows the expression of the transgene protein (Glut1-GFP) after transfection in human brain microvascular endothelial cells (hCMEC / d3).

[0152] Figure 10A. GFP fluorescence 72 hours after transfection using a construct containing one of several endothelial cell promoters that drive the expression of the Glut1-GFP transgene.

[0153] Figure 10B. GFP fluorescence 72 hours after transfection using a construct containing one of the following: two ubiquitous promoters (CMV or CAG), a Glut1-free control vector (CMV-GFP), or no transfection (no NFX). Images acquired using Operetta CLS® (PerkinElmer®).

[0154] Figure 10C. This figure shows an expression cassette containing the target promoter (hFLT1, mTie, hTie, or hGlut1), the GLUT1 (SLC2A1) gene (T2A link-GFP), and regulatory elements adjacent to the AAV2 inverted terminal repeat (ITR).

[0155] Figures 11A-11C. 2-deoxy-D-glucose (glucose) uptake in hCMEC / d3 cells after expression of human GLUT1 (SLC2A1). Human brain microvascular endothelial cells (hCMEC / d3) were transfected with plasmids expressing the hGLUT1-t2A-eGFP transgene via CAG-GFP (CON; negative control) or several endothelium-specific promoters (i.e., hFLT1, mTie, hTie, or hGlut1) or ubiquitous CMV or CAG promoters. Glucose uptake was measured using a luminescence-based kit (Promega®) with 0.5 mM 2-deoxyglucose (2-DG) in culture medium. Glucose (2-DG) uptake was normalized across all cells using phase-contrast imaging [error bars represent SEM; n=6 replications per state].

[0156] Figure 11A. Glucose (2-DG) uptake was measured 72 hours after transfection in the first experiment.

[0157] Figure 11B. Glucose (2-DG) uptake was measured 72 hours after transfection in the second experiment.

[0158] Figure 11C. Glucose (2-DG) uptake was measured 96 hours after transfection.

[0159] Figure 12A. Glucose (2-DG) uptake in hCMEC / D3 cells at 72 hours after human Glut1 (SLC2A1) expression.

[0160] Figure 12B. Glucose (2-DG) uptake at 96 hours after expression of human Glut1 (SLC2A1) in hCMEC / D3 cells.

[0161] Figure 13. Shows the uptake of 2-deoxy-D-glucose (glucose) associated with the expression of hGLUT1 (SLC2A1) via AAV9 in hCMEC / D3 cells. Human brain microvascular endothelial cells (hCMEC / d3) were subjected to an AAV9 vector expressing either CAG-GFP (negative control) (3x10 5 Transduction was performed using a vector genome / cell, or an hGLUT1 transgene driven by one of several endothelial-specific promoters (i.e., hFLT1, mTie1, or hGlut1), or by a ubiquitous CMV promoter. Glucose (2-DG) uptake was measured 72 hours post-transduction using a luminescence-based Glucose Uptake-Glo kit (Promega®) and normalized per cell using the RealTime-Glo MT cell viability assay (Promega®) [error bars represent SEM; n=4 replications per state].

[0162] Example 3: In vivo evaluation of AAV9-mediated GLUT1 expression using an endothelial promoter in a GLUT1-deficient animal model. A series of experiments will be conducted to evaluate the in vivo effects of AAV9-mediated expression of the Glut1 transporter protein in a GLUT1 deficiency syndrome (DS) model mouse. This model uses heterozygous haploinsufficiency mice (GLUT-1 + / - mice) with targeted disruption of the mouse GLUT-1 gene promoter and exon 1 region, exhibiting characteristics of human GLUT DS, such as paroxysmal syndrome, hypoglycemia, microcephaly, and motor dysfunction (Wang et al, Hum Mol Gen, 2006; Tang et al., Nat Comm, 2016). The AAV9 construct will express the GLUT1 transgene via either a ubiquitous promoter (CMV) or one of several endothelial cell promoters (hFLT-1, mTie, hGlut1) and will be evaluated at different doses and administration routes (intravenous or intraventricular). After administration with the AAV9 vector, the extent to which endothelial cell promoter-mediated GLUT1 transgene expression can prevent or mitigate functional and pathological impairments in the model mouse will be evaluated. The potential beneficial effects of AAV9-mediated Glut1 protein expression administered to heterozygous haploinsufficient mice were revealed in comparison with untreated GLUT-1 + / - control mice, and included improved or normalized weight gain, behavioral performance in exercise tests (e.g., rotation rod, vertical pole evaluation), CSF glucose levels, brain weight, and the integrity and size of cerebral microvascular structure (e.g., cerebral capillary density, vessel size, number of branching points, etc.).

[0163] Sequence information SEQUENCE LISTING <110> Spacecraft Seven, LLC <120> ADENO-ASSOCIATED VIRAL VECTOR FOR GLUT1 EXPRESSION AND USES THEREOF <150> US 63 / 061,726 <151> 2020-08-05 <160> 102 <170> PatentIn version 3.5 <210> 1 <211> 1037 <212> DNA <213> Homo sapiens <400> 1 tttgcttcta ggagcaga ggactgagga atgacttggg cgggtgcatc aatgcggcca aaaaagacac ggacacgctc ccctggggacc tgagctggtt cgcagtcttc ccaaaggtgc caagcaagcg tcagttcccc tcaggcgctc caggttcagt gccttgtgcc gagggtctcc ggtgccttcc tagacttctc gggacagtct gaaggggtca ggagcggcgg gacagcgcgg 240 gaagagcagg caagggaga cagccggact gcgcctcagt cctccgtgcc aagaacaccg tcgcggaggc gcggccagct tcccttggat cggactttcc gcccctaggg ccaggcggcg 360 gagcttcagc cttgtccctt ccccagtttc gggcggcccc cagagctgag tagccgggt 420 ggaggggagtc tgcaaggatt tcctgagcgc gatgggcagg aggaggggca agggcaagag 480 ggcgcggagc aaagaccctg aacctgccgg ggccgcgctc ccggggcccgc gtcgccagca 540 cctccccacg cgcgctcggc cccggggcc cggccctcgt cggcccccgc ccctctccgt 600. agccgcaggg aagcgagcct gggaggaaga agagggtagg tggggaggcg gatgaggggt 660 gggggacccc ttgacgtcac cagaaggagg tgccggggta ggaagtgggc tggggaaagg 720 ttaaatcg cccccgccct cggctgctct tcatcgaggt ccgcgggagg ctcggagcgc 780 gccaggcgga cactcctctc ggctcctcc cggcagcggc ggcggctcgg agcgggctcc 840 ggggctcggg tgcagcggcc agcgggcgcc tggcggcgag gattacccgg ggaagtggtt 900 gtctcctggc tggagccgcg agacgggcgc tcagggcgg gggccggcgg cggcgaacaa 960 gaggacggac tctggcggcc gggtcgttgg ccgcggggag cgcgggcacc gggcgagcag 1020 gccgcgtcgc gctcacc 1037 <210> 2 <211> 1608 <212> DNA <213> Homo sapiens <400> 2 agctcctccc agcctcaggc ccaggaatgg gaatctctgt gggtcacaca tcagtaggga 60 ggtctttccc gatccttttc tatgctactc caggagtcaa agcgtctcct gggactttc 120 agggcgcttc agaagagccc tgggcctaaa ccagctcaac caagctgcag ggacccagcc 180 tcctgagaaa agtgaatgtg agccccggtgc attcagagga gaatgaagcc ttcacccaga 240 acacatctg ggaagatgtc ccaggcccag ggggagggtt tgtactacca gacctaagtc 300 acctaaactg acaccaagtc tcatccatcc caaccattcc attccgggtc agaggggtca 360 tcgatttaac cagcaaggct gcccatccaa cggttgctcc ctctgctccc tggaagggcc 420 tcctcgtggg cgttctgtac ctacaggtct tgttccgttc tgggaactgc cagtggtggc 480 aagaggtgga gcaacgggtg ccagggcagg gagaggtgag tctgggaggg aagcagaggc 540 aagatccatg gggctttaga gactttgcca aagcagtgcg actgctccca ggttgttgtc 600 agccgtcaag agtgagtgca cctccctggg cagacttctg ctgccccagt gcccaggaat 660 aggcaggggt ttgccgcaaa atgaatgaca cctggcagac aataagctga agctttcatt 720 agcagcttaa gctgaggact atctatgcaa ccgatactcc ctgtgtgctc cccgggactg 780 cttaatgtga gcccttgtgg agcgattggc accaagaaag caaggactaa gtcagaagtt 840 caagtcccag ccttgccaca gcctcagggt gccctcgagc acagcaagcc tcagttttcc 900 catctgtaca atgagagagg tacacaaggt agactcgaag gctctttgtt gccagggccc 960 tgtgttcctt tgagtgtatg tgcttctcag gcccacagag gtcctttgtg tttcgtatgt 1020 gaactgctct ctaggaaacc catgtaactg tctgtgtcct ggggcacata catgaggact 1080 catgtgggcc gtattgtgtg tttgtgccgg ggggagggga gaccccagaa caatgtcccc 1140 caccccaccc ccctcctcaa taggcggaag ccactggctt cctccctttc ctgcctcctg 1200 cctcctttgt gccagcaaga ctgagtactg gagagagaca ggggatggga aaaatcagtc 1260 cagctgtccc caggtctgcc cttaccataa ccttcccccc acctcaagtg actcctccca 1320 ggccacaccc atccccagcc ttgtgggggc cagattgggg ggcctagagg ctcaaaggca 1380 gaatgagtcc tcccaccccc taccctgcca cccctcccac ccaagccacc tcatttcctc 1440 ttcctcccca gcaccgaccc acactgacca acacaggctg agcagtcagg cccacagcat 1500 ctgaccccag gcccagctcg tcctggctgg cctgggtcgg cctctggagt atggtctggc 1560 gggtgccccc tttcttgctc cccatcctct tcttggcttc tcatgtgg 1608 <210> 3 <211> 2510 <212> DNA <213> Homo sapiens <400> 3 60. ctagtagcag aaacaaggtc ctctggaaga gcaactgatg ctcttaggta ctgaagcatc atcctgcccc agagaccact cgcatatgaa gcacacatat tcagtctgcc ttacttgtgt taatgattgc cagtgtccct ctgacctcct agccctgaaa agtgtggcct gaaggtcatt tcagagacgg ggagagctgc tcagagaagc caatcggcga gtctaggaca cacagacagg atctagtccc agctagtcgct agcctagtg agcgtcccct ggccccttat accacttcct 300 tctccagctt gcatctaatc tgctctggca gaccatcgtg tttcctgtct tcctggcagc 360 420. ctccagcacg ctcagtgcta ctccctgcgc atgcgccctc tccagtgggc ttggagtgcg aggaggagg gtgaggagg ggtgaaatca ggtattggat 480. ccacaggggg tctgaagagc actagcctgg cctttgggga ctgaacttct gctatgaaga 540 cctccactgc catccctgga gtccggggca catccaaggc ttgctgtcca tcgtttactg 600 tttacagatg acaacaatga ctgtgttcgg ggcagaaata tccaccaggg ctagagtaca 660 aaaggagttt gcattgatgg ccggacaggc cctgtccctg gcagcctgcc agcgctgagt 720 atgagaccca gcgggaagtg ctaccctggc agacgtgtcc actgagtaca cagaccacca 780 aggcaggcag ctctcgggga agctgtctat gctgggccag cccaccttga gggcagggaa 840 cagaacagat tgtggcagag aggaaaatgt ggagcttctg tttgttcaca gacacacgca 900 ctcgcccacg cacgcacgca cgcacgcacg cacgcacgaa tgcacgcacg cagtagttga 960 atgctatgga ttccgctcag agctgagaac agccccagcg acagttccct ggcctctctc 1020 cttactctga tgtcctcatc tgtcttcaca tggtctcagg acgctaatac tccatcctaa 1080 tgtacactcc tttccctggg cctccgttcc agttcagttc tcagaggacc tggagggagt 1140 gattggctac accaactttg ctttcgttca ccaagcccat gtctctactt gggtgtctaa 1200 tgggcatctc caacattacc taccccaaac agaaaaccct ttcttccccc caaccacacc 1260 ccaccctacc cccacagtat tttctccatg cccggaaaga tctgctctct tatggtccct 1320 ctttgcctca ctgaaaagca ggacaagttg gggacttccc aaacttttat gcatgaagaa 1380 acccaggcaa tttgccaaaa ggtacactct gggggtctgt catttactct gagccagaac 1440 cctgaaattt ttactaaccc atcacataat gaatgaagag aatctttttc tttttttttt 1500 tttttctttt tttttggttt ttcgagacag ggtttctctg tatagccctg gctatcctgg 1560 aacacactct gtagaccagg ctggcctcga actcagaaat ccacctgcct ctgcctcccg 1620 agtgctggga ttaaaggcgt gcgccaccac gcctggctga atgaagagaa tcttgacctc 1680 atctccccag cctcttggtc ctgagggacc ctggtctacc tactgctttg ctgtcttctt 1740 agctcttctt acttttttgc tgactcagac ctatggctat ctccattata cagatgagga 1800 gactgaggca tggatccctg gttggtccat ggtcacgtga agcccatcac ccagtatttg 1860 taaagtgaga tgggccaggc tggtaccttg gaactgaaac tcacactgcc ctacctggaa 1920 gaatctgaca ggcaaaatct gctgctgaaa gtgattgtct gtcacgtttc tcagctgccc 1980 gactctgaga actccacagc cccctttcgt tccaccatac tacagagtcg ccacggaaag 2040 ccggctctgt ggagaagctg aggtagctgg gtttctgtct gggttactct gtccagcgag 2100 gaaacaagta ccttagaccc actaagcctc tgctttctga actgtaaagt gggggatatg 2160 acacctgcct cccagggatg gctgaatgct ctggcagaag cttagagccc ccacagctac 2220 ccctaggctc acagctcctc cgatgagacc tagaattgag gtatgagttg aataccccag 2280 gcaggtccaa ggcttccacg ggcccaggct gaccaagctg aggccgccca ccgtagggct 2340 tgcctatctg caggcagctc acaaaggaac aataacagga aaccatcccg aggggaagtg 2400 ggccagggcc agttggaaaa cctgcctccc tcccagcctg ggtgtggctc cctctcccc 2460 tcctgaggca atcaactgtg ctctccacaa agctcggccc tggacagact 2510 <210> 4 <211> 94 <212> DNA <213> Homo sapiens <400> 4 gctggagcct cggtagccgt tcctcctgcc cgctgggcct cccaacgggc cctcctccc 60 tccttgcacc ggcccttcct ggtctttgaa taaa 94 <210> 5 <211> 1476 <212> DNA <213> Homo sapiens <400> 5 atggagccca gcagcaagaa gctgacgggt cgcctcatgc tggccgtggg aggagcagtg cttggctccc tgcagtttgg ctacaacact ggagtcatca atgcccccca gaaggtgatc 180. gaggagttct acaaccagac atgggtccac cgctatgggg agagcatcct gcccaccacg ctcaccacgc tctggtccct ctcagtggcc atcttttctg ttgggggcat gattggctcc 240 ttctctgtgg gccttttcgt taaccgcttt ggccggcgga attcaatgct gatgatgaac ctgctggcct tcgtgtccgc cgtgctcatg ggcttctcga aactgggcaa gtcctttgag 360 atgctgatcc tgggccgctt catcatcggt gtgtactgcg gcctgaccac aggcttcgtg 420 cccatgtatg tgggtgaagt gtcacccaca gcccttcgtg gggccctggg caccctgcac 480 cagctgggca tcgtcgtcgg catcctcatc gcccaggtgt tcggcctgga ctccatcatg 540 ggcaacaagg acctgtggcc cctgctgctg agcatcatct tcatcccggc cctgctgcag tgcatcgtgc tgcccttctg ccccgagagt ccccgcttcc tgctcatcaa ccgcaacgag 660 gagaaccggg ccaagagtgt gctaaagaag ctgcgcggga cagctgacgt gacccatgac 720 ctgcaggaga tgaaggaaga gagtcggcag atgatgcggg agaagaaggt caccatcctg 780 gagctgttcc gctcccccgc ctaccgccag cccatcctca tcgctgtggt gctgcagctg 840 tcccagcagc tgtctggcat caacgctgtc ttctattact ccacgagcat cttcgagaag 900 gcgggggtgc agcagcctgt gtatgccacc attggctccg gtatcgtcaa cacggccttc 960 actgtcgtgt cgctgtttgt ggtggagcga gcaggccggc ggaccctgca cctcataggc 1020 ctcgctggca tggcgggttg tgccatactc atgaccatcg cgctagcact gctggagcag 1080 ctaccctgga tgtcctatct gagcatcgtg gccatctttg gctttgtggc cttctttgaa 1140 gtgggtcctg gccccatccc atggttcatc gtggctgaac tcttcagcca gggtccacgt 1200 ccagctgcca ttgccgttgc aggcttctcc aactggacct caaatttcat tgtgggcatg 1260 tgcttccagt atgtggagca actgtgtggt ccctacgtct tcatcatctt cactgtgctc 1320 ctggttctgt tcttcatctt cacctacttc aaagttcctg agactaaagg ccggaccttc 1380 gatgagatcg cttccggctt ccggcagggg ggagccagcc aaagtgacaa gacacccgag 1440 gagctgttcc atcccctggg ggctgattcc caagtg 1476 <210> 6 <211> 168 <212> DNA <213> Adeno-associated virus 2 <400> 6 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 120 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgta 168 <210> 7 <211> 168 <212> DNA <213> Adeno-associated virus 2 <400> 7 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag 60 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 120 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgc 168 <210> 8 <211> 2963 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - part of expression cassette <400> 8 ctctggagac gcgttacata cgttacataa cttacggtaa atggcccgcc tggctgaccg 60 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata 120 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta 180 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc 240 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac 300 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga 360 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg 420 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg 480 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac 540 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac 600 cgatccagcc tccgcggatg gagcccagca gcaagaagct gacgggtcgc ctcatgctgg 660 ccgtgggagg agcagtgctt ggctccctgc agtttggcta caacactgga gtcatcaatg 720 ccccccagaa ggtgatcgag gagttctaca accagacatg ggtccaccgc tatggggaga 780 gcatcctgcc caccacgctc accacgctct ggtccctctc agtggccatc ttttctgttg 840 ggggcatgat tggctccttc tctgtgggcc ttttcgttaa ccgctttggc cggcggaatt 900 caatgctgat gatgaacctg ctggccttcg tgtccgccgt gctcatgggc ttctcgaaac 960 tgggcaagtc ctttgagatg ctgatcctgg gccgcttcat catcggtgtg tactgcggcc 1020 tgaccacagg cttcgtgccc atgtatgtgg gtgaagtgtc acccacagcc cttcgtgggg 1080 ccctgggcac cctgcaccag ctgggcatcg tcgtcggcat cctcatcgcc caggtgttcg 1140 gcctggactc catcatgggc aacaaggacc tgtggcccct gctgctgagc atcatcttca 1200 tcccggccct gctgcagtgc atcgtgctgc ccttctgccc cgagagtccc cgcttcctgc 1260 tcatcaaccg caacgaggag aaccgggcca agagtgtgct aaagaagctg cgcgggacag 1320 ctgacgtgac ccatgacctg caggagatga aggaagagag tcggcagatg atgcgggaga 1380 agaaggtcac catcctggag ctgttccgct cccccgccta ccgccagccc atcctcatcg 1440 ctgtggtgct gcagctgtcc cagcagctgt ctggcatcaa cgctgtcttc tattactcca 1500 cgagcatctt cgagaaggcg ggggtgcagc agcctgtgta tgccaccatt ggctccggta 1560 tcgtcaacac ggccttcact gtcgtgtcgc tgtttgtggt ggagcgagca ggccggcgga 1620 ccctgcacct cataggcctc gctggcatgg cgggttgtgc catactcatg accatcgcgc 1680 tagcactgct ggagcagcta ccctggatgt cctatctgag catcgtggcc atctttggct 1740 ttgtggcctt ctttgaagtg ggtcctggcc ccatcccatg gttcatcgtg gctgaactct 1800 tcagccaggg tccacgtcca gctgccattg ccgttgcagg cttctccaac tggacctcaa 1860 atttcattgt gggcatgtgc ttccagtatg tggagcaact gtgtggtccc tacgtcttca 1920 tcatcttcac tgtgctcctg gttctgttct tcatcttcac ctacttcaaa gttcctgaga 1980 ctaaaggccg gaccttcgat gagatcgctt ccggcttccg gcagggggga gccagccaaa 2040 gtgacaagac acccgaggag ctgttccatc ccctgggggc tgattcccaa gtgtgataat 2100 ggatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc 2160 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg 2220 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt 2280 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac 2340 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc 2400 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct 2460 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct 2520 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct 2580 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct 2640 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcattgcct 2700 gcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac 2760 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt 2820 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc ccaagttggg 2880 aagaaacctg tagggcctgc gttacccagg ctggagtgca gtggcacatt tctgctcact 2940 gcaacctcct cctccctggg ttc 2963 <210> 9 <400> 9 000 <210> 10 <211> 3414 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - part of expression cassette <400> 10 ctctggagac gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac 60 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc 120 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg 180 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat 240 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc 300 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc 360 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg 420 gcgggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc 480 gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat 540 ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc 600 tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct 660 ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg 720 taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga 780 ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt 840 gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg 900 cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc 960 cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg 1020 ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc 1080 ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc gtggcgcggg 1140 gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc 1200 tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg gcggctgtcg 1260 aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg cgcagggact 1320 tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca ccccctctag 1380 cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg agggccttcg 1440 tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc cgcgggggga 1500 cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg 1560 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa 1620 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcatggagc ccagcagcaa 1680 gaagctgacg ggtcgcctca tgctggccgt gggaggagca gtgcttggct ccctgcagtt 1740 tggctacaac actggagtca tcaatgcccc ccagaaggtg atcgaggagt tctacaacca 1800 gacatgggtc caccgctatg gggagagcat cctgcccacc acgctcacca cgctctggtc 1860 cctctcagtg gccatctttt ctgttggggg catgattggc tccttctctg tgggcctttt 1920 cgttaaccgc tttggccggc ggaattcaat gctgatgatg aacctgctgg ccttcgtgtc 1980 cgccgtgctc atgggcttct cgaaactggg caagtccttt gagatgctga tcctgggccg 2040 cttcatcatc ggtgtgtact gcggcctgac cacaggcttc gtgcccatgt atgtgggtga 2100 agtgtcaccc acagcccttc gtggggccct gggcaccctg caccagctgg gcatcgtcgt 2160 cggcatcctc atcgcccagg tgttcggcct ggactccatc atgggcaaca aggacctgtg 2220 gcccctgctg ctgagcatca tcttcatccc ggccctgctg cagtgcatcg tgctgccctt 2280 ctgccccgag agtccccgct tcctgctcat caaccgcaac gaggagaacc gggccaagag 2340 tgtgctaaag aagctgcgcg ggacagctga cgtgacccat gacctgcagg agatgaagga 2400 agagagtcgg cagatgatgc gggagaagaa ggtcaccatc ctggagctgt tccgctcccc 2460 cgcctaccgc cagcccatcc tcatcgctgt ggtgctgcag ctgtcccagc agctgtctgg 2520 catcaacgct gtcttctatt actccacgag catcttcgag aaggcggggg tgcagcagcc 2580 tgtgtatgcc accattggct ccggtatcgt caacacggcc ttcactgtcg tgtcgctgtt 2640 tgtggtggag cgagcaggcc ggcggaccct gcacctcata ggcctcgctg gcatggcggg 2700 ttgtgccata ctcatgacca tcgcgctagc actgctggag cagctaccct ggatgtccta 2760 tctgagcatc gtggccatct ttggctttgt ggccttcttt gaagtgggtc ctggccccat 2820 cccatggttc atcgtggctg aactcttcag ccagggtcca cgtccagctg ccattgccgt 2880 tgcaggcttc tccaactgga cctcaaattt cattgtgggc atgtgcttcc agtatgtgga 2940 gcaactgtgt ggtccctacg tcttcatcat cttcactgtg ctcctggttc tgttcttcat 3000 cttcacctac ttcaaagttc ctgagactaa aggccggacc ttcgatgaga tcgcttccgg 3060 cttccggcag gggggagcca gccaaagtga caagacaccc gaggagctgt tccatcccct 3120 gggggctgat tcccaagtgt gatcattgcc tgcccgggtg gcatccctgt gacccctccc 3180 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa 3240 aattaagttg catcattttg tctgactagg tgtccttcta tatattatg gggtggaggg 3300 gggtggtatg gagcaagggg cccaagttgg gaagaaacct gtagggcctg cgttacccag 3360 gctggagtgc agtggcacat ttctgctcac tgcaacctcc tcctccctgg gttc 3414 <210> 11 <400> 11 000 <210> 12 <211> 3409 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - part of expression cassette <400> 12 ctctggagac gcgttacata tttgcttcta ggaagcagaa gactgaggaa atgacttggg 60 cgggtgcatc aatgcggcca aaaaagacac ggacacgctc ccctgggacc tgagctggtt 120 cgcagtcttc ccaaaggtgc caagcaagcg tcagttcccc tcaggcgctc caggttcagt 180 gccttgtgcc gagggtctcc ggtgccttcc tagacttctc gggacagtct gaaggggtca 240 ggagcggcgg gacagcgcgg gaagagcagg caaggggaga cagccggact gcgcctcagt 300 cctccgtgcc aagaacaccg tcgcggaggc gcggccagct tcccttggat cggactttcc 360 gcccctaggg ccaggcggcg gagcttcagc cttgtccctt ccccagtttc gggcggcccc 420 cagagctgag taagccgggt ggagggagtc tgcaaggatt tcctgagcgc gatgggcagg 480 aggaggggca agggcaagag ggcgcggagc aaagaccctg aacctgccgg ggccgcgctc 540 ccgggcccgc gtcgccagca cctccccacg cgcgctcggc cccgggccac ccgccctcgt 600 cggccccgc ccctctccgt agccgcaggg aagcgagcct gggaggaaga agaggtagg 660 tgggggaggcg gatgaggggt gggggacccc ttgacgtcac cagaaggagg tgccggggta 720 ggaagtgggc tggggaaagg ttataaatcg cccccgccct cggctgctct tcatcgaggt 780 ccgcgggagg ctcggagcgc gccaggcgga cactcctctc ggctcctccc cggcagcggc 840 ggcggctcgg agcgggctcc ggggctcggg tgcagcggcc agcgggcgcc tggcggcgag 900 gattacccgg ggaagtggtt gtctcctggc tggagccgcg agacgggcgc tcagggcgcg 960 gggccggcgg cggcgaacaa gaggacggac tctggcggcc gggtcgttgg ccgcggggag 1020 cgcgggcacc gggcgagcag gccgcgtcgc gctcaccgcc accatggagc ccagcagcaa 1080 gaagctgacg ggtcgcctca tgctggccgt gggaggagca gtgcttggct ccctgcagtt 1140 tggctacaac actggagtca tcaatgcccc ccagaaggtg atcgaggagt tctacaacca 1200 gacatgggtc caccgctatg gggagagcat cctgcccacc acgctcacca cgctctggtc 1260 cctctcagtg gccatctttt ctgttggggg catgattggc tccttctctg tgggcctttt 1320 cgttaaccgc tttggccggc ggaattcaat gctgatgatg aacctgctgg ccttcgtgtc 1380 cgccgtgctc atgggcttct cgaaactggg caagtccttt gagatgctga tcctgggccg 1440 cttcatcatc ggtgtgtact gcggcctgac cacaggcttc gtgcccatgt atgtgggtga 1500 agtgtcaccc acagcccttc gtggggccct gggcaccctg caccagctgg gcatcgtcgt 1560 cggcatcctc atcgcccagg tgttcggcct ggactccatc atgggcaaca aggacctgtg 1620 gcccctgctg ctgagcatca tcttcatccc ggccctgctg cagtgcatcg tgctgccctt 1680 ctgccccgag agtccccgct tcctgctcat caaccgcaac gaggagaacc gggccaagag 1740 tgtgctaaag aagctgcgcg ggacagctga cgtgacccat gacctgcagg agatgaagga 1800 agagagtcgg cagatgatgc gggagaagaa ggtcaccatc ctggagctgt tccgctcccc 1860 cgcctaccgc cagcccatcc tcatcgctgt ggtgctgcag ctgtcccagc agctgtctgg 1920 catcaacgct gtcttctatt actccacgag catcttcgag aaggcggggg tgcagcagcc 1980 tgtgtatgcc accattggct ccggtatcgt caacacggcc ttcactgtcg tgtcgctgtt 2040 tgtggtggag cgagcaggcc ggcggaccct gcacctcata ggcctcgctg gcatggcggg 2100 ttgtgccata ctcatgacca tcgcgctagc actgctggag cagctaccct ggatgtccta 2160 tctgagcatc gtggccatct ttggctttgt ggccttcttt gaagtgggtc ctggccccat 2220 cccatggttc atcgtggctg aactcttcag ccagggtcca cgtccagctg ccattgccgt 2280 tgcaggcttc tccaactgga cctcaaattt cattgtgggc atgtgcttcc agtatgtgga 2340 gcaactgtgt ggtccctacg tcttcatcat cttcactgtg ctcctggttc tgttcttcat 2400 cttcacctac ttcaaagttc ctgagactaa aggccggacc ttcgatgaga tcgcttccgg 2460 cttccggcag gggggagcca gccaaagtga caagacaccc gaggagctgt tccatcccct 2520 gggggctgat tcccaagtgt gataatggat caacctctgg attacaaaat ttgtgaaaga 2580 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg 2640 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc 2700 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc 2760 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt 2820 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt 2880 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg 2940 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg 3000 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg 3060 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt 3120 tgggccgcct ccccgcatca ttgcctgccc gggtggcatc cctgtgaccc ctccccagtg 3180 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta 3240 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg 3300 gtatggagca aggggcccaa gttgggaaga aacctgtagg gcctgcgtta cccaggctgg 3360 agtgcagtgg cacatttctg ctcactgcaa cctcctcctc cctgggttc 3409 <210> 13 <400> 13 000 <210> 14 <211> 3980 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - part of expression cassette <400> 14 ctctggagac gcgttacata agctcctccc agcctcaggc ccaggaatgg gaatctctgt 60 gggtcacaca tcagtaggga ggtctttccc gatccttttc tatgctactc caggagtcaa 120 agcgtctcct gggacttttc agggcgcttc agaagagccc tgggcctaaa ccagctcaac 180 caagctgcag ggacccagcc tcctgagaaa agtgaatgtg agcccggtgc attcagagga 240 gaatgaagcc ttcacccaga acacactctg ggaagatgtc ccaggcccag ggggagggtt 300 tgtactacca gacctaagtc acctaaactg acaccaagtc tcatccatcc caaccattcc 360 attccgggtc agaggggtca tcgatttaac cagcaaggct gcccatccaa cggttgctcc 420 ctctgctccc tggaagggcc tcctcgtggg cgttctgtac ctacaggtct tgttccgttc 480 tgggaactgc cagtggtggc aagaggtgga gcaacgggtg ccagggcagg gagaggtgag 540 tctgggagg aagcagaggc aagatccatg gggctttaga gactttgcca aagcagtgcg 600 actgctccca ggttgttgtc agccgtcaag agtgagtgca cctccctggg cagacttctg 660 ctgccccagt gcccaggaat aggcaggggt ttgccgcaaa atgaatgaca cctggcagac 720 aataagctga agctttcatt agcagcttaa gctgaggact atctatgcaa ccgatactcc 780 ctgtgtgctc cccgggactg cttaatgtga gcccttgtgg agcgattggc accaagaaag 840 caaggactaa gtcagaagtt caagtcccag ccttgccaca gcctcagggt gccctcgagc 900 acagcaagcc tcagttttcc catctgtaca atgagagagg tacaaaggt agactcgaag 960 gctctttgtt gccagggccc tgtgttcctt tgagtgtatg tgcttctcag gcccacagag 1020 gtcctttgtg tttcgtatgt gaactgctct ctaggaaacc catgtaactg tctgtgtcct 1080 ggggcacata catgaggact catgtgggcc gtattgtgtg tttgtgccgg ggggagggga 1140 gaccccagaa caatgtcccc caccccaccc ccctcctcaa taggcggaag ccactggctt 1200 cctccctttc ctgcctcctg cctcctttgt gccagcaaga ctgagtactg gagagagaca 1260 ggggatggga aaaatcagtc cagctgtccc caggtctgcc cttaccataa ccttcccccc 1320 acctcaagtg actcctccca ggccacaccc atccccagcc ttgtgggggc cagattgggg 1380 ggcctagagg ctcaaaggca gaatgagtcc tcccaccccc taccctgcca cccctcccac 1440 ccaagccacc tcatttcctc ttcctcccca gcaccgaccc acactgacca acacaggctg 1500 agcagtcagg cccacagcat ctgaccccag gcccagctcg tcctggctgg cctgggtcgg 1560 cctctggagt atggtctggc gggtgccccc tttcttgctc cccatcctct tcttggcttc 1620 tcatgtgggc caccatggag cccagcagca agaagctgac gggtcgcctc atgctggccg 1680 tgggaggac agtgcttggc tccctgcagt ttggctacaa cactggagtc atcaatgccc 1740 cccagaaggt gatcgaggag ttctacaacc agacatgggt ccaccgctat ggggagagca 1800 tcctgcccac cacgctcacc acgctctggt ccctctcagt ggccatcttt tctgttgggg 1860 gcatgattgg ctccttctct gtgggccttt tcgttaaccg ctttggccgg cggaattcaa 1920 tgctgatgat gaacctgctg gccttcgtgt ccgccgtgct catgggcttc tcgaaactgg 1980 gcaagtcctt tgagatgctg atcctgggcc gcttcatcat cggtgtgtac tgcggcctga 2040 ccacaggctt cgtgcccatg tatgtgggtg aagtgtcacc cacagccctt cgtggggccc 2100 tgggcaccct gcaccagctg ggcatcgtcg tcggcatcct catcgcccag gtgttcggcc 2160 tggactccat catgggcaac areacctgt ggcccctgct gctgagcatc atcttcatcc 2220 cggccctgct gcagtgcatc gtgctgccct tctgccccga gagtccccgc ttcctgctca 2280 tcaaccgcaa cgaggagaac cgggccaaga gtgtgctaaa gaagctgcgc gggacagctg 2340 acgtgaccca tgacctgcag gagatgaagg aagagagtcg ccagatgatg cgggaagaa 2400 aggtcaccat cctggagctg ttccgctccc ccgcctaccg ccagcccatc ctcatcgctg 2460 tggtgctgca gctgtcccag cagctgtctg gcatcaacgc tgtcttctat tactccacga 2520 gcatcttcga gaaggcgggg gtgcagcagc ctgtgtatgc caccattggc tccggtatcg 2580 tcaacacggc cttcactgtc gtgtcgctgt ttgtggtgga gcgagcaggc cggcggaccc 2640 tgcacctcat aggcctcgct ggcatggcgg gttgtgccat actcatgacc atcgcgctag 2700 cactgctgga gcagctaccc tggatgtcct atctgagcat cgtggccatc tttggctttg 2760 tggccttctt tgaagtgggt cctggcccca tcccatggtt catcgtggct gaactcttca 2820 gccagggtcc acgtccagct gccattgccg ttgcaggctt ctccaactgg acctcaaatt 2880 tcattgtggg catgtgcttc cagtatgtgg agcaactgtg tggtccctac gtcttcatca 2940 tcttcactgt gctcctggtt ctgttcttca tcttcaccta cttcaaagtt cctgagacta 3000 aaggccggac cttcgatgag atcgcttccg gcttccggca ggggggagcc agccaaagtg 3060 acaagacacc cgaggagctg ttccatcccc tgggggctga ttcccaagtg tgataatgga 3120 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 3180 tttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 3240 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 3300 gccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 3360 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc cccccctat 3420 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 3480 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tctttcctt ggctgctcgc 3540 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 3600 tccagcggac cttccttccc gcggcctgct gcggctctg cggcctcttc cgcgtcttcg 3660 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc attgcctgcc 3720 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc 3780 agtgcccacc agccttgtcc tataaaatt aagttgcatc atttgtctg actaggtgtc 3840 cttctataat attatggggt ggaggggggt ggtatggagc aaggggccca agttgggaag 3900 aaacctgtag ggcctgcgtt acccaggctg gagtgcagtg gcacatttct gctcactgca 3960 acctcctcct ccctgggttc 3980 <210> 15 <400> 15 000 <210> 16 <211> 4380 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - part of expression cassette <400> 16 ctctggagac gcgttacata ctagtagcag aaacaaggtc ctctggaaga gcaactgatg 60 ctcttaggta ctgaagcatc atcctgcccc agagaccact cgcatatgaa gcacacatat 120 tcagtctgcc ttacttgtgt taatgattgc cagtgtccct ctgacctcct agccctgaaa 180 agtgtggcct gaaggtcatt tcagagacgg ggagagctgc tcagagaagc caatcggcga 240 gtctaggaca cacagacagg atctagtccc agagttcgct agcctaggtg agcgtcccct 300 ggccccttat accacttcct tctccagctt gcatctaatc tgctctggca gaccatcgtg 360 tttcctgtct tcctggcagc ctccagcacg ctcagtgcta ctccctgcgc atgcgccctc 420 ctcccagtac cttctctgac tccagtgggc ttggagtgcg aggaggaagg gtgaggaagg 480 ggtgaaatca ggtattggat ccacaggggg tctgaagagc actagcctgg ccttttggga 540 ctgaacttct gctatgaaga cctccactgc catccctgga gtccggggca catccaaggc 600 ttgctgtcca tcgtttactg tttacagatg acaacaatga ctgtgttcgg ggcagaaata 660 tccaccaggg ctagagtaca aaaggagttt gcattgatgg ccggacaggc cctgtccctg 720 gcagcctgcc agcgctgagt atgagaccca gcgggaagtg ctaccctggc agacgtgtcc 780 actgagtaca cagaccacca aggcaggcag ctctcgggga agctgtctat gctgggccag 840 cccaccttga gggcagggaa cagaacagat tgtggcagag aggaaaatgt ggagcttctg 900 tttgttcaca gacacacgca ctcgcccacg cacgcacgca cgcacgcacg cacgcacgaa 960 tgcacgcacg cagtagttga atgctatgga ttccgctcag agctgagaac agccccagcg 1020 acagttccct ggcctctctc cttactctga tgtcctcatc tgtcttcaca tggtctcagg 1080 acgctaatac tccatcctaa tgtacactcc tttccctggg cctccgttcc agttcagttc 1140 tcagaggacc tggagggagt gattggctac accaactttg ctttcgttca ccaagcccat 1200 gtctctactt gggtgtctaa tgggcatctc caacattacc taccccaaac agaaaaccct 1260 ttcttccccc caaccacacc ccaccctacc cccacagtat tttctccatg cccggaaaga 1320 tctgctctct tatggtccct ctttgcctca ctgaaaagca ggacaagttg gggacttccc 1380 aaacttttat gcatgaagaa acccaggcaa tttgccaaaa ggtacactct gggggtctgt 1440 catttactct gagccagaac cctgaaattt ttactaaccc atcacataat gaatgaagag 1500 aatctttttc tttttttttt tttttctttt tttttggttt ttcgagacag ggtttctctg 1560 tatagccctg gctatcctgg aacacactct gtagaccagg ctggcctcga actcagaaat 1620 ccacctgcct ctgcctcccg agtgctggga ttaaaggcgt gcgccaccac gcctggctga 1680 atgaagagaa tcttgacctc atctccccag cctcttggtc ctgagggacc ctggtctacc 1740 tactgctttg ctgtcttctt agctcttctt acttttttgc tgactcagac ctatggctat 1800 ctccattata cagatgagga gactgaggca tggatccctg gttggtccat ggtcacgtga 1860 agcccatcac ccagtatttg taaagtgaga tgggccaggc tggtaccttg gaactgaaac 1920 tcacactgcc ctacctggaa gaatctgaca ggcaaaatct gctgctgaaa gtgattgtct 1980 gtcacgtttc tcagctgccc gactctgaga actccacagc cccctttcgt tccaccatac 2040 tacagagtcg ccacggaaag ccggctctgt ggagaagctg aggtagctgg gtttctgtct 2100 gggttactct gtccagcgag gaaacaagta ccttagaccc actaagcctc tgctttctga 2160 actgtaaagt gggggatatg acacctgcct cccagggatg gctgaatgct ctggcagaag 2220 cttagagccc ccacagctac ccctaggctc acagctcctc cgatgagacc tagaattgag 2280 gtatgagttg aataccccag gcaggtccaa ggcttccacg ggcccaggct gaccaagctg 2340 aggccgccca ccgtagggct tgcctatctg caggcagctc acaaaggaac aataacagga 2400 aaccatcccg aggggaagtg ggccagggcc agttggaaaa cctgcctccc tcccagcctg 2460 ggtgtggctc ccctctcccc tcctgaggca atcaactgtg ctctccacaa agctcggccc 2520 tggacagact gccaccatgg agcccagcag caagaagctg acgggtcgcc tcatgctggc 2580 cgtgggagga gcagtgcttg gctccctgca gtttggctac aacactggag tcatcaatgc 2640 cccccagaag gtgatcgagg agttctacaa ccagacatgg gtccaccgct atggggagag 2700 catcctgccc accacgctca ccacgctctg gtccctctca gtggccatct tttctgttgg 2760 gggcatgatt ggctccttct ctgtgggcct tttcgttaac cgctttggcc ggcggaattc 2820 aatgctgatg atgaacctgc tggccttcgt gtccgccgtg ctcatgggct tctcgaaact 2880 gggcaagtcc tttgagatgc tgatcctggg ccgcttcatc atcggtgtgt actgcggcct 2940 gaccacaggc ttcgtgccca tgtatgtggg tgaagtgtca cccacagccc ttcgtggggc 3000 cctgggcacc ctgcaccagc tgggcatcgt cgtcggcatc ctcatcgccc aggtgttcgg 3060 cctggactcc atcatgggca acaaggacct gtggcccctg ctgctgagca tcatcttcat 3120 cccggccctg ctgcagtgca tcgtgctgcc cttctgcccc gagagtcccc gcttcctgct 3180 catcaaccgc aacgaggaga accgggccaa gagtgtgcta aagaagctgc gcgggacagc 3240 tgacgtgacc catgacctgc aggagatgaa ggaagagagt cggcagatga tgcgggagaa 3300 gaaggtcacc atcctggagc tgttccgctc ccccgcctac cgccagccca tcctcatcgc 3360 tgtggtgctg cagctgtccc agcagctgtc tggcatcaac gctgtcttct attactccac 3420 gagcatcttc gagaaggcgg gggtgcagca gcctgtgtat gccaccattg gctccggtat 3480 cgtcaacacg gccttcactg tcgtgtcgct gtttgtggtg gagcgagcag gccggcggac 3540 cctgcacctc ataggcctcg ctggcatggc gggttgtgcc atactcatga ccatcgcgct 3600 agcactgctg gagcagctac cctggatgtc ctatctgagc atcgtggcca tctttggctt 3660 tgtggccttc tttgaagtgg gtcctggccc catcccatgg ttcatcgtgg ctgaactctt 3720 cagccagggt ccacgtccag ctgccattgc cgttgcaggc ttctccaact ggacctcaaa 3780 tttcattgtg ggcatgtgct tccagtatgt ggagcaactg tgtggtccct acgtcttcat 3840 catcttcact gtgctcctgg ttctgttctt catcttcacc tacttcaaag ttcctgagac 3900 taaaggccgg accttcgatg agatcgcttc cggcttccgg caggggggag ccagccaaag 3960 tgacaagaca cccgaggagc tgttccatcc cctgggggct gattcccaag tgtgagctgg 4020 agcctcggta gccgttcctc ctgcccgctg ggcctcccaa cgggccctcc tcccctcctt 4080 gcaccggccc ttcctggtct ttgaataaac attgcctgcc cgggtggcat ccctgtgacc 4140 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc 4200 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt 4260 ggaggggggt ggtatggagc aaggggccca agttgggaag aaacctgtag ggcctgcgtt 4320 acccaggctg gagtgcagtg gcacatttct gctcactgca acctcctcct ccctgggttc 4380 <210> 17 <211> 3299 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - full polynucleotide sequence of vector genome <400> 17 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 120 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtact ctggagacgc 180 gttacatacg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 240 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 300 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 360 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 420 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 480 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 540 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 600 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 660 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc 720 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc 780 cgcggatgga gcccagcagc aagaagctga cgggtcgcct catgctggcc gtgggaggag 840 cagtgcttgg ctccctgcag tttggctaca acactggagt catcaatgcc ccccagaagg 900 tgatcgagga gttctacaac cagacatggg tccaccgcta tggggagagc atcctgccca 960 ccacgctcac cacgctctgg tccctctcag tggccatctt ttctgttggg ggcatgattg 1020 gctcctttc tgtgggcctt ttcgttaacc gctttggccg gcggaattca atgctgatga 1080 tgaacctgct ggccttcgtg tccgccgtgc tcatgggctt ctcgaaactg ggcaagtcct 1140 ttgagatgct gatcctgggc cgcttcatca tcggtgtgta ctgcggcctg accacaggct 1200 tcgtgcccat gtatgtgggt gaagtgtcac ccacagccct tcgtggggcc ctgggcaccc 1260 tgcaccagct gggcatcgtc gtcggcatcc tcatcgccca ggtgttcggc ctggactcca 1320 tcatgggcaa caaggacctg tggcccctgc tgctgagcat catcttcatc ccggccctgc 1380 tgcagtgcat cgtgctgccc ttctgccccg agagtccccg cttcctgctc atcaaccgca 1440 agaggagaa ccgggccaag agtgtgctaa agaagctgcg cgggacagct gacgtgaccc 1500 atgacctgca ggagatgaag gaagagagtc ggcagatgat gcgggagaag aaggtcacca 1560 tcctggagct gttccgctcc cccgcctacc gccagcccat cctcatcgct gtggtgctgc 1620 agctgtccca gcagctgtct ggcatcaacg ctgtcttcta ttactccacg agcatcttcg 1680 agaaggcggg ggtgcagcag cctgtgtatg ccaccattgg ctccggtatc gtcaacacgg 1740 ccttcactgt cgtgtcgctg tttgtggtgg agcgagcagg ccggcggacc ctgcacctca 1800 taggcctcgc tggcatggcg ggttgtgcca tactcatgac catcgcgcta gcactgctgg 1860 agcagctacc ctggatgtcc tatctgagca tcgtggccat ctttggcttt gtggccttct 1920 ttgaagtggg tcctggcccc atcccatggt tcatcgtggc tgaactcttc agccagggtc 1980 cacgtccagc tgccattgcc gttgcaggct tctccaactg gacctcaaat ttcattgtgg 2040 gcatgtgctt ccagtatgtg gagcaactgt gtggtcccta cgtcttcatc atcttcactg 2100 tgctcctggt tctgttcttc atcttcacct acttcaaagt tcctgagact aaaggccgga 2160 ccttcgatga gatcgcttcc ggcttccggc aggggggagc cagccaaagt gacaagacac 2220 ccgaggagct gttccatccc ctgggggctg attcccaagt gtgataatgg atcaacctct 2280 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct 2340 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat 2400 tttctctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt 2460 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca accccccactg gttggggcat 2520 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc 2580 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga 2640 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc 2700 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga 2760 ccttcttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttgccc 2820 tcagacgagt cggatctccc tttgggccgc ctccccgcat cattgcctgc ccgggtggca 2880 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac 2940 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa 3000 tattatgggg tggagggggg tggtatggag caaggggccc aagttgggaa gaaacctgta 3060 gggcctgcgt tacccaggct ggagtgcagt ggcacatttc tgctcactgc aacctcctcc 3120 tccctgggtt ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc 3180 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac 3240 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgc 3299 <210> 18 <400> 18 000 <210> 19 <211> 3750 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - full polynucleotide sequence of vector genome <400> 19 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 120 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtact ctggagacgc 180 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 240 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 300 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 360 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 420 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 480 atggtcgagg tgagccccac gttctgcttc actctcccca tctcccccccc ctccccaccc 540 ccaattttgt atttattat tttttaatta ttttgtgcag cgatggggc gggggggggg 600 ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg 660 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg 720 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct 780 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc 840 gttatccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt 900 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga 960 gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag 1020 cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct 1080 ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg 1140 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg 1200 gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga 1260 gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc 1320 gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga 1380 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag 1440 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca 1500 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc 1560 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg 1620 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg 1680 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc 1740 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat 1800 tgtgctgtct catcattttg gcaaagaatt catggagccc agcagcaaga agctgacggg 1860 tcgcctcatg ctggccgtgg gaggagcagt gcttggctcc ctgcagtttg gctacaacac 1920 tggagtcatc aatgcccccc agaaggtgat cgaggagttc tacaaccaga catgggtcca 1980 ccgctatggg gagagcatcc tgcccaccac gctcaccacg ctctggtccc tctcagtggc 2040 catcttttct gttgggggca tgattggctc cttctctgtg ggccttttcg ttaaccgctt 2100 tggccggcgg aattcaatgc tgatgatgaa cctgctggcc ttcgtgtccg ccgtgctcat 2160 gggcttctcg aaactgggca agtcctttga gatgctgatc ctgggccgct tcatcatcgg 2220 tgtgtactgc ggcctgacca caggcttcgt gcccatgtat gtgggtgaag tgtcacccac 2280 agcccttcgt ggggccctgg gcaccctgca ccagctgggc atcgtcgtcg gcatcctcat 2340 cgcccaggtg ttcggcctgg actccatcat gggcaacaag gacctgtggc ccctgctgct gagcatcatc ttcatcccgg ccctgctgca gtgcatcgtg ctgcccttct gccccgagag 2460 tccccgcttc ctgctcatca accgcaacga ggagaaccgg gccaagagtg tgctaaaga gctgcgcggg acagctgacg tgacccatga cctgcaggag atgaaggaag agagtcggca 2640. gatgatgcgg gagaagaagg tcaccatcct ggagctgttc cgctcccccg cctaccgcca gcccatcctc atcgctgtgg tgctgcagct gtcccagcag ctgtctggca tcaacgctgt 2700 cttctattac tccacgagca tcttcgagaa ggcggggggtg cagcagcctg tgtatgccac cattggctcc ggtatcgtca acacggcctt cactgtcgtg tcgctgtttg tggtggagcg 2820 agcaggccgg cggaccctgc acctcatagg cctcgctggc atggcgggtt gtgccatact 2880 catgaccatc gcgctagcac tgctggagca gctaccctgg atgtcctatc tgagcatcgt ggccatcttt ggctttgtgg ccttctttga agtgggtcct ggccccatcc catggttcat3000 cgtggctgaa ctcttcagcc agggtccacg tccagctgcc attgccgttg caggcttctc 3060 caactggacc tcaaatttca ttgtgggcat gtgcttccag tatgtggagc aactgtgtgg 3120 tccctacgtc ttcatcatct tcactgtgct cctggttctg ttcttcatct tcacctactt 3180 caaagttcct gagactaaag gccggacctt cgatgagatc gcttccggct tccggcaggg 3240 gggagccagc caaagtgaca agaacccga ggagctggttc catcccctgg gggctgattc 3300 ccaagtgtga tcattgcctg cccgggtggc atccctgtga cccctcccca gtgcctctcc 3360 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca 3420 tcattttgtc tgactaggtg tcctttata atattatggg gtggaggggg gtggtatgga 3480 gcaaggggcc caagttggga agaaacctgt agggcctgcg ttacccaggc tggagtgcag 3540 tggcacattt ctgctcactg caacctcctc ctccctgggt tctacgtaga tagtagcat 3600 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg 3660 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc 3720 cgggcggcct cagtgagcga gcgagcgcgc 3750 <210> 20 <400> 20 000 <210> 21 <211> 3745 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - full polynucleotide sequence of vector genome <400> 21 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 120 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtact ctggagacgc 180 gttacatatt tgcttctagg aagcagaaga ctgaggaaat gacttgggcg ggtgcatcaa 240 tgcggccaaa aaagacacgg acacgctccc ctgggacctg agctggttcg cagtcttccc 300 aaaggtgcca agcaagcgtc agttcccctc aggcgctcca ggttcagtgc cttgtgccga 360 gggtctccgg tgccttccta gacttctcgg gacagtctga aggggtcagg agcggcggga 420 cagcgcggga agagcaggca aggggagaca gccggactgc gcctcagtcc tccgtgccaa 480 gaacaccgtc gcggaggcgc ggccagcttc ccttggatcg gactttccgc ccctagggcc 540 aggcggcgga gcttcagcct tgtcccttcc ccagtttcgg gcggccccca gagctgagta 600 agccgggtgg agggagtctg caaggatttc ctgagcgcga tgggcaggag gaggggcaag 660 ggcaagaggg cgcggagcaa agaccctgaa cctgccgggg ccgcgctccc gggcccgcgt 720 cgccagcacc tccccacgcg cgctcggccc cgggccaccc gccctcgtcg gcccccgccc 780 ctctccgtag ccgcagggaa gcgagcctgg gaggaagaag agggtaggtg gggaggcgga 840 tgaggggtgg gggacccctt gacgtcacca gaaggaggtg ccggggtagg aagtgggctg 900 gggaaaggtt ataaatcgcc cccgccctcg gctgctcttc atcgaggtcc gcgggaggct 960 cggagcgcgc caggcggaca ctcctctcgg ctcctccccg gcagcggcgg cggctcggag 1020 cgggctccgg ggctcgggtg cagcggccag cgggcgcctg gcggcgagga ttacccgggg 1080 aagtggttgt ctcctggctg gagccgcgag acgggcgctc agggcgcggg gccggcggcg 1140 gcgaacaaga ggacggactc tggcggccgg gtcgttggcc gcggggagcg cgggcaccgg 1200 gcgagcaggc cgcgtcgcgc tcaccgccac catggagccc agcagcaaga agctgacggg 1260 tcgcctcatg ctggccgtgg gaggagcagt gcttggctcc ctgcagtttg gctacaacac 1320 tggagtcatc aatgcccccc agaaggtgat cgaggagttc tacaaccaga catgggtcca 1380 ccgctatggg gagagcatcc tgcccaccac gctcaccacg ctctggtccc tctcagtggc 1440 catcttttct gttgggggca tgattggctc cttctctgtg ggccttttcg ttaaccgctt 1500 tggccggcgg aattcaatgc tgatgatgaa cctgctggcc ttcgtgtccg ccgtgctcat 1560 gggcttctcg aaactgggca agtcctttga gatgctgatc ctgggccgct tcatcatcgg 1620 tgtgtactgc ggcctgacca caggcttcgt gcccatgtat gtgggtgaag tgtcacccac 1680 agcccttcgt ggggccctgg gcaccctgca ccagctgggc atcgtcgtcg gcatcctcat 1740 cgcccaggtg ttcggcctgg actccatcat gggcaacaag gacctgtggc ccctgctgct 1800 gagcatcatc ttcatcccgg ccctgctgca gtgcatcgtg ctgcccttct gccccgagag 1860 tccccgcttc ctgctcatca accgcaacga ggagaaccgg gccaagagtg tgctaaagaa 1920 gctgcgcggg acagctgacg tgacccatga cctgcaggag atgaaggaag agagtcggca 1980 gatgatgcgg gagaagaagg tcaccatcct ggagctgttc cgctcccccg cctaccgcca gcccatcctc atcgctgtgg tgctgcagct gtcccagcag ctgtctggca tcaacgctgt 2100 cttctattac tccacgagca tcttcgagaa ggcggggggtg cagcagcctg tgtatgccac cattggctcc ggtatcgtca acacggcctt cactgtcgtg tcgctgtttg tggtggagcg 2220 agcaggccgg cggaccctgc acctcatagg cctcgctggc atggcgggtt gtgccatact 2280 catgaccatc gcgctagcac tgctggagca gctaccctgg atgtcctatc tgagcatcgt ggccatcttt ggctttgtgg ccttctttga agtgggtcct ggccccatcc catggttcat2400 cgtggctgaa ctcttcagcc agggtccacg tccagctgcc attgccgttg caggcttctc caactggacc tcaaatttca ttgtgggcat gtgcttccag tatgtggagc aactgtgtgg tccctacgtc ttcatcatct tcactgtgct cctggttctg ttcttcatct tcacctactt caaagttcct gagactaaag gccggacctt cgatgagatc gcttccggct tccggcaggg gggagccagc caaagtgaca agacacccga ggagctgttc catcccctgg gggctgattc ccaagtgtga taatggatca acctctggat tacaaaattt gtgaaagatt gactggtatt 2760 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 2820 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 2880 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 2940 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 3000 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 3060 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc 3120 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 3180 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 3240 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 3300 ccgcatcatt gcctgcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc 3360 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt 3420 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag 3480 gggcccaagt tgggaagaaa cctgtagggc ctgcgttacc caggctggag tgcagtggca 3540 catttctgct cactgcaacc tcctcctccc tgggttctac gtagataagt agcatggcgg 3600 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc 3660 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc 3720 ggcctcagtg agcgagcgag cgcgc 3745 <210> 22 <400> 22 000 <210> 23 <211> 4316 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - full polynucleotide sequence of vector genome <400> 23 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 120 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtact ctggagacgc 180 gttacataag ctcctcccag cctcaggccc aggaatggga atctctgtgg gtcacacatc 300. tctttcccga tccttttcta tgctactcca ggagtcaaag cgtctcctgg gacttttcag ggcgcttcag aagagccctg ggcctaacc agctcaacca agctgcaggg 360 acccagcctc ctgagaaaag tgaatgtgag cccggtgcat tcagaggaga atgaagcctt 420 cacccagaac acactctggg aagatgtccc aggcccaggg ggaggggtttg tactaccaga 480 540. ctaagtcac ctaaactgac accaagtctc atccatccca accattccat tccgggtcag aggggtcatc gatttaacca gcaaggctgc ccatccaacg gttgctccct ctgctccctg gaagggcctc ctcgtggggcg ttctgtacct acaggtcttg ttccgttctg ggaactgcca 660 gtggtggcaa gaggtggagc aacgggtgcc agggcaggga gaggtgagtc tgggaggga 720. gcagaggcaa gatccatggg gctttagaga ctttgccaaa gcagtgcgac tgctcccagg 780 ttgttgtcag ccgtcaagag tgagtgcacc tccctgggca gacttctgct gccccagtgc 840 ccaggaatag gcaggggttt gccgcaaaat gaatgacacc tggcagacaa tagctgaag ctttcattag cagcttaagc tgaggactat ctatgcaacc gatactccct gtgtgctccc 960 cgggactgct taatgtgagc ccttgtggag cgattggcac caagaaagca aggactaagt 1020 cagaagttca agtcccagcc ttgccacagc ctcagggtgc cctcgagcac agcaagcctc 1080 agttttccca tctgtacaat gagagaggta cacaaggtag actcgaaggc tctttgttgc 1140 cagggccctg tgttcctttg agtgtatgtg cttctcaggc ccacagaggt cctttgtgtt 1200 tcgtatgtga actgctctct aggaaaccca tgtaactgtc tgtgtcctgg ggcacataca 1260 tgaggactca tgtgggccgt attgtgtgtt tgtgccgggg ggaggggaga ccccagaaca 1320 atgtccccca ccccaccccc ctcctcaata ggcggaagcc actggcttcc tccctttcct 1380 gcctcctgcc tcctttgtgc cagcaagact gagtactgga gagagacagg ggatgggaaa 1440 aatcagtcca gctgtcccca ggtctgccct taccataacc ttccccccac ctcaagtgac 1500 tcctcccagg ccacacccat ccccagcctt gtgggggcca gattgggggg cctagaggct 1560 caaaggcaga atgagtcctc ccacccccta ccctgccacc cctcccaccc aagccacctc 1620 atttcctctt cctccccagc accgacccac actgaccaac acaggctgag cagtcaggcc 1680 cacagcatct gaccccaggc ccagctcgtc ctggctggcc tgggtcggcc tctggagtat 1740 ggtctggcgg gtgccccctt tcttgctccc catcctcttc ttggcttctc atgtgggcca 1800 ccatggagcc cagcagcaag aagctgacgg gtcgcctcat gctggccgtg ggaggagcag 1860 tgcttggctc cctgcagttt ggctacaaca ctggagtcat caatgccccc cagaaggtga 1920 tcgaggagtt ctacaaccag acatgggtcc accgctatgg ggagagcatc ctgcccacca 1980 cgctcaccac gctctggtcc ctctcagtgg ccatcttttc tgttgggggc atgattggct 2040 ccttctctgt gggccttttc gttaaccgct ttggccggcg gaattcaatg ctgatgatga 2100 acctgctggc cttcgtgtcc gccgtgctca tgggcttctc gaaactgggc aagtcctttg 2160 agatgctgat cctgggccgc ttcatcatcg gtgtgtactg cggcctgacc acaggcttcg 2220 tgcccatgta tgtgggtgaa gtgtcaccca cagcccttcg tggggccctg ggcaccctgc 2280 accagctggg catcgtcgtc ggcatcctca tcgcccaggt gttcggcctg gactccatca 2340 tgggcaacaa ggacctgtgg cccctgctgc tgagcatcat cttcatcccg gccctgctgc agtgcatcgt gctgcccttc tgccccgaga gtccccgctt cctgctcatc aaccgcaacg 2460 aggagaaccg ggccaagagt gtgctaaaga agctgcgcgg gacagctgac gtgacccatg acctgcagga gatgagga gagagtcggc agatgatgcg gtcaccatcc tggagctgtt ccgctccccc gcctaccgcc agcccatcct catcgctgtg gtgctgcagc 2640 tgtcccagca gctgtctggc atcaacgctg tcttctatta ctccacgagc atcttcgaga 2700 aggcgggggt gcagcagcct gtgtatgcca ccattggctc cggtatcgtc aacacggcct 2760 tcactgtcgt gtcgctgttt gtggtggagc gagcaggccg gcggaccctg cacctcatag 2820 gcctcgctgg catggcgggt tgtgccatac tcatgaccat cgcgctagca ctgctggagc 2880 agctaccctg gatgtcctat ctgagcatcg tggccatctt tggctttgtg gccttctttg 2940 aagtgggtcc tggccccatc ccatggttca tcgtggctga actcttcagc cagggtccac 3060. gtccagctgc cattgccgtt gcaggcttct ccaactggac ctcaaatttc attgtgggca tgtgcttcca gtatgtggag caactgtgtg gtccctacgt cttcatcatc ttcactgtgc 3120 tcctggttct gttcttcatc ttcacctact tcaaagttcc tgagactaaa ggccggacct 3180 tcgatgagat cgcttccggc ttccggcagg ggggagccag ccaaagtgac aagacacccg 3240 aggagctgtt ccatcccctg ggggctgatt cccaagtgtg ataatggatc aacctctgga 3300 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 3360 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 3420 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 3480 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 3540 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 3600 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 3660 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac 3720 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 3780 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 3840 gacgagtcgg atctcccttt gggccgcctc cccgcatcat tgcctgcccg ggtggcatcc 3900 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 3960 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 4020 tatggggtgg aggggggtgg tatggagcaa ggggcccaag ttgggaagaa acctgtaggg 4080 cctgcgttac ccaggctgga gtgcagtggc acatttctgc tcactgcaac ctcctcctcc 4140 ctgggttcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag 4200 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa 4260 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgc 4316 <210> 24 <400> 24 000 <210> 25 <211> 4716 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - full polynucleotide sequence of vector genome <400> 25 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 120 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtact ctggagacgc 180 gttacatact agtagcagaa acaaggtcct ctggaagagc aactgatgct cttaggtact 240 gaagcatcat cctgccccag agaccactcg catatgaagc acacatattc agtctgcctt 300 acttgtgtta atgattgcca gtgtccctct gacctcctag ccctgaaaag tgtggcctga 360 aggtcatttc agagacgggg agagctgctc agagaagcca atcggcgagt ctaggacaca 420 cagacaggat ctagtcccag agttcgctag cctaggtgag cgtcccctgg ccccttatac 480 cacttccttc tccagcttgc atctaatctg ctctggcaga ccatcgtgtt tcctgtcttc 540 ctggcagcct ccagcacgct cagtgctact ccctgcgcat gcgccctcct cccagtacct 600 tctctgactc cagtgggctt ggagtgcgag gaggaagggt gaggaagggg tgaaatcagg 660 tattggatcc acagggggtc tgaagagcac tagcctggcc ttttgggact gaacttctgc 720 tatgaagacc tccactgcca tccctggagt ccggggcaca tccaaggctt gctgtccatc 780 gtttactgtt tacagatgac aacaatgact gtgttcgggg cagaaatatc caccagggct 840 agagtacaaa aggagtttgc attgatggcc ggacaggccc tgtccctggc agcctgccag 900 cgctgagtat gagacccagc gggaagtgct accctggcag acgtgtccac tgagtacaca 960 gaccaccaag gcaggcagct ctcggggaag ctgtctatgc tgggccagcc caccttgagg 1020 gcagggaaca gaacagattg tggcagagag gaaaatgtgg agcttctgtt tgttcacaga 1080 cacacgcact cgcccacgca cgcacgcacg cacgcacgca cgcacgaatg cacgcacgca 1140 gtagttgaat gctatggatt ccgctcagag ctgagaacag ccccagcgac agttccctgg 1200 cctctctcct tactctgatg tcctcatctg tcttcacatg gtctcaggac gctaatactc 1260 catcctaatg tacactcctt tccctgggcc tccgttccag ttcagttctc agaggacctg 1320 gagggagtga ttggctacac caactttgct ttcgttcacc aagcccatgt ctctacttgg 1380 gtgtctaatg ggcatctcca acattaccta ccccaaacag aaaacccttt cttcccccca 1440 accacacccc accctacccc cacagtattt tctccatgcc cggaaagatc tgctctctta 1500 tggtccctct ttgcctcact gaaaagcagg acaagttggg gacttcccaa acttttatgc 1560 atgaagaaac ccaggcaatt tgccaaaagg tacactctgg gggtctgtca tttactctga 1620 gccagaaccc tgaaattttt actaacccat cacataatga atgaagagaa tctttttctt 1680 tttttttt tttcttttt tttggtttt cgagacaggg tttctctgta tagccctggc 1740 tatcctggaa cacactctgt agaccaggct ggcctcgaac tcagaaatcc acctgcctct 1800 gcctcccgag tgctgggatt aaaggcgtgc gccaccacgc ctggctgaat gaagagaatc 1860 ttgacctcat ctccccagcc tcttggtcct gagggaccct ggtctaccta ctgctttgct 1920 gtcttcttag ctcttcttac ttttttgctg actcagacct atggctatct ccattataca 1980 gatgaggaga ctgaggcatg gatccctggt tggtccatgg tcacgtgaag cccatcaccc 2040 agtatttgta aagtgagatg ggccaggctg gtaccttgga actgaaactc acactgccct 2100 acctggaaga atctgacagg caaaatctgc tgctgaaagt gattgtctgt cacgtttctc 2160 agctgcccga ctctgagaac tccacagccc cctttcgttc caccatacta cagagtcgcc 2220 acggaaagcc ggctctgtgg agaagctgag gtagctgggt ttctgtctgg gttactctgt 2280 ccagcgagga aacaagtacc ttagacccac taagcctctg ctttctgaac tgtaaagtgg 2340 gggatatgac acctgcctcc cagggatggc tgaatgctct ggcagaagct tagagccccc 2400 acagctaccc ctaggctcac agctcctccg atgagaccta gaattgaggt atgagttgaa 2460 taccccaggc aggtccaagg cttccacggg cccaggctga ccaagctgag gccgcccacc 2520 gtagggcttg cctatctgca ggcagctcac aaaggaacaa taacaggaaa ccatcccgag 2580 gggaagtggg ccagggccag ttggaaaacc tgcctccctc ccagcctggg tgtggctccc 2640 ctctcccctc ctgaggcaat caactgtgct ctccacaaag ctcggccctg gacagactgc 2700 caccatggag cccagcagca agaagctgac gggtcgcctc atgctggccg tgggaggagc 2760 agtgcttggc tccctgcagt ttggctacaa cactggagtc atcaatgccc cccagaaggt 2820 gatcgaggag ttctacaacc agacatgggt ccaccgctat ggggagagca tcctgcccac 2880 cacgctcacc acgctctggt ccctctcagt ggccatcttt tctgttgggg gcatgattgg 2940 ctccttctct gtgggccttt tcgttaaccg ctttggccgg cggaattcaa tgctgatgat 3000 gaacctgctg gccttcgtgt ccgccgtgct catgggcttc tcgaaactgg gcaagtcctt 3060 tgagatgctg atcctgggcc gcttcatcat cggtgtgtac tgcggcctga ccacaggctt 3120 cgtgcccatg tatgtgggtg aagtgtcacc cacagccctt cgtggggccc tgggcaccct 3180 gcaccagctg ggcatcgtcg tcggcatcct catcgcccag gtgttcggcc tggactccat 3240 catgggcaac aaggacctgt ggcccctgct gctgagcatc atcttcatcc cggccctgct 3300 gcagtgcatc gtgctgccct tctgccccga gagtccccgc ttcctgctca tcaaccgcaa 3360 cgaggagaac cgggccaaga gtgtgctaaa gaagctgcgc gggacagctg acgtgaccca 3420 tgacctgcag gagatgaagg aagagagtcg gcagatgatg cgggagaaga aggtcaccat 3480 cctggagctg ttccgctccc ccgcctaccg ccagcccatc ctcatcgctg tggtgctgca 3540 gctgtcccag cagctgtctg gcatcaacgc tgtcttctat tactccacga gcatcttcga 3600 gaaggcgggg gtgcagcagc ctgtgtatgc caccattggc tccggtatcg tcaacacggc 3660 cttcactgtc gtgtcgctgt ttgtggtgga gcgagcaggc cggcggaccc tgcacctcat 3720 aggcctcgct ggcatggcgg gttgtgccat actcatgacc atcgcgctag cactgctgga 3780 gcagctaccc tggatgtcct atctgagcat cgtggccatc tttggctttg tggccttctt 3840 tgaagtgggt cctggcccca tcccatggtt catcgtggct gaactcttca gccagggtcc 3900 acgtccagct gccattgccg ttgcaggctt ctccaactgg acctcaaatt tcattgtggg 3960 catgtgcttc cagtatgtgg agcaactgtg tggtccctac gtcttcatca tcttcactgt 4020 gctcctggtt ctgttcttca tcttcaccta cttcaaagtt cctgagacta aaggccggac 4080 cttcgatgag atcgcttccg gcttccggca ggggggagcc agccaaagtg acaagacacc 4140 cgaggagctg ttccatcccc tgggggctga ttcccaagtg tgagctggag cctcggtagc 4200 cgttcctcct gcccgctggg cctcccaacg ggccctcctc ccctccttgc accggccctt 4260 cctggtcttt gaataaacat tgcctgcccg ggtggcatcc ctgtgacccc tccccagtgc 4320 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa 4380 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg 4440 tatggagcaa ggggcccaag ttgggaagaa acctgtaggg cctgcgttac ccaggctgga 4500 gtgcagtggc acatttctgc tcactgcaac ctcctcctcc ctgggttcta cgtagataag 4560 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc 4620 tctctgcgcg ctgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgc 4680 tttgcccggg cggcctcagt gagcgagcga gcgcgc 4716 <210> 26 <211> 492 <212> PRT <213> Homo sapiens <400> 26 Put Glu Pro Ser Ser Lys Lys Leu Thr Gly Arg Leu Put Leu Ala Val 1 5 10 15 Gly Gly Ala Val Leu Gly Ser Leu Gln Phe Gly Tyr Asn Thr Gly Val 20 25 30 Ile Asn Ala Pro Gln Lys Val Ile Glu Glu Phe Tyr Asn Gln Thr Trp 35 40 45 Val His Arg Tyr Gly Glu Ser Ile Leu Pro Thr Thr Leu Thr Thr Leu 50 55 60 Trp Ser Leu Ser Val Ala Ile Phe Ser Val Gly Gly Met Ile Gly Ser 65 70 75 80 Phe Ser Val Gly Leu Phe Val Asn Arg Phe Gly Arg Arg Asn Ser Met 85 90 95 Leu Met Met Asn Leu Leu Ala Phe Val Ser Ala Val Leu Met Gly Phe 100 105 110 Ser Lys Leu Gly Lys Ser Phe Glu Met Leu Ile Leu Gly Arg Phe Ile 115 120 125 Ile Gly Val Tyr Cys Gly Leu Thr Thr Gly Phe Val Pro Met Tyr Val 130 135 140 Gly Glu Val Ser Pro Thr Ala Leu Arg Gly Ala Leu Gly Thr Leu His 145 150 155 160 Gln Leu Gly Ile Val Val Gly Ile Leu Ile Ala Gln Val Phe Gly Leu 165 170 175 Asp Ser Ile Met Gly Asn Lys Asp Leu Trp Pro Leu Leu Leu Ser Ile 180 185 190 Ile Phe Ile Pro Ala Leu Leu Gln Cys Ile Val Leu Pro Phe Cys Pro 195 200 205 Glu Ser Pro Arg Phe Leu Leu Ile Asn Arg Asn Glu Glu Asn Arg Ala 210 215 220 Lys Ser Val Leu Lys Lys Leu Arg Gly Thr Ala Asp Val Thr His Asp 225 230 235 240 Leu Gln Glu Met Lys Glu Glu Ser Arg Gln Met Met Arg Glu Lys Lys 245 250 255 Val Thr Ile Leu Glu Leu Phe Arg Ser Pro Ala Tyr Arg Gln Pro Ile 260 265 270 Leu Ile Ala Val Val Leu Gln Leu Ser Gln Gln Leu Ser Gly Ile Asn 275 280 285 Ala Val Phe Tyr Tyr Ser Thr Ser Ile Phe Glu Lys Ala Gly Val Gln 290 295 300 Gln Pro Val Tyr Ala Thr Ile Gly Ser Gly Ile Val Asn Thr Ala Phe 305 310 315 320 Thr Val Val Ser Leu Phe Val Val Glu Arg Ala Gly Arg Arg Thr Leu 325 330 335 His Leu Ile Gly Leu Ala Gly Met Ala Gly Cys Ala Ile Leu Met Thr 340 345 350 Ile Ala Leu Ala Leu Leu Glu Gln Leu Pro Trp Met Ser Tyr Leu Ser 355 360 365 Ile Val Ala Ile Phe Gly Phe Val Ala Phe Phe Glu Val Gly Pro Gly 370 375 380 Pro Ile Pro Trp Phe Ile Val Ala Glu Leu Phe Ser Gln Gly Pro Arg 385 390 395 400 Pro Ala Ala Ile Ala Val Ala Gly Phe Ser Asn Trp Thr Ser Asn Phe 405 410 415 Ile Val Gly Met Cys Phe Gln Tyr Val Glu Gln Leu Cys Gly Pro Tyr 420 425 430 Val Phe Ile Ile Phe Thr Val Leu Leu Val Leu Phe Phe Ile Phe Thr 435 440 445 Tyr Phe Lys Val Pro Glu Thr Lys Gly Arg Thr Phe Asp Glu Ile Ala 450 455 460 Ser Gly Phe Arg Gln Gly Gly Ala Ser Gln Ser Asp Lys Thr Pro Glu 465 470 475 480 Glu Leu Phe His Pro Leu Gly Ala Asp Ser Gln Val 485 490 <210> 27 <211> 1476 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - Codon-optimized polynucleotide encoding GLUT1 <400> 27 atggaaccat catccaaaaa gctgaccgga cgactgatgc ttgcagttgg cggtgcggtc ttggggagcc tgcagtttgg gtacaatact ggcgtaatca atgccccgca gaaggttatt gagaatttt acaatcaaac gtgggtacat cgctacggtg aatccattct tcctacaact ctgaccacac tctggagcct ttctgtagcg attttttccg tcgggggcat gataggatca ttttccgtcg gtctttttgt gaaccgcttt ggccggagaa attccatgct gatgatgaat cttctcgctt tcgtgagtgc cgtcctcatg ggatttagta aactgggtaa atctttcgag 360 atgttgatac tggggagatt tattatcggc gtgtattgtg gtttgaccac gggctttgta 420 ccaatgtatg ttggcgaggt ttctccgaca gcattgagag gtgcactcgg gaccttgcac 480 cagttgggca tcgtagtag aatccttata gcgcaagttt tcgggctcga ttccatcatg 540. gggaacaaag atctctggcc attgctcctc tcaataattt ttataccggc attgcttcag tgtattgttc ttcctttttg cccagagtcc cctaggttcc tgctcataaa caggaatgag 660 gagaatcgcg ctaagtccgt gttgaaaaaa cttaggggaa ctgcagacgt tactcacgat 720 ttgcaagaga tgaaggagga atctaggcaa atgatgcgcg agaagaaggt taccatactc 780 gaactcttcc gctcccccgc gtacaggcag cccattctta tcgcggtcgt cttgcagttg 840 tcacaacagt tgagtgggat taatgcagtt ttctattata gcacgtccat atttgaaaaa 900 gcaggcgtcc aacaacctgt ctatgcaact ataggctcag gcattgtaaa cacagcgttt 960 actgtagtat cactgtttgt cgttgagcgg gctggtcgaa ggaccttgca cctcatagga 1020 ctggcgggca tggcgggctg tgcgattctt atgacaattg cgctcgcgct gttggaacag 1080 cttccgtgga tgtcctatct ctctatagta gcaatatttg gatttgttgc attttttgaa 1140 gttgggcccg gacctatccc ctggttcatc gtcgcggagc tcttttccca aggcccaaga 1200 ccggctgcca ttgctgttgc aggcttctca aactggacga gtaatttcat agtaggtatg 1260 tgtttccagt atgttgaaca gctctgtggg ccctatgtct ttatcatctt tactgtgttg 1320 ctcgtgttgt tctttatctt cacttatttc aaagtacccg agacaaaggg caggacgttt 1380 gacgagattg catctggttt tagacaagga ggtgcctcac agagtgataa aaccccggag 1440 gaattgtttc atccgctggg agccgactca caggtc 1476 <210> 28 <211> 10 <212> DNA <213> Artificial Sequence <220> <223> Kozak sequence motif <400> 28 gccaccatgg 10 <210> 29 <211> 1482 <212> DNA <213> Artificial Sequence <220> <223> Polynucleotide encoding GLUT1 with Kozak motif <400> 29 gccaccatgg agcccagcag caagaagctg acgggtcgcc tcatgctggc cgtgggagga 60 gcagtgcttg gctccctgca gtttggctac aacactggag tcatcaatgc cccccagaag 120 gtgatcgagg agttctacaa ccagacatgg gtccaccgct atggggagag catcctgccc 180 accacgctca ccacgctctg gtccctctca gtggccatct tttctgttgg gggcatgatt 240 ggctccttct ctgtgggcct tttcgttaac cgctttggcc ggcggaattc aatgctgatg 300. atgaacctgc tggccttcgt gtccgccgtg ctcatgggct tctcgaaact gggcaagtcc 360 tttgagatgc tgatcctggg ccgcttcatc atcggtgtgt actgcggcct gaccacaggc 420 ttcgtgccca tgtatgtggg tgaagtgtca cccacagccc ttcgtggggc cctgggcacc 480 ctgcaccagc tgggcatcgt cgtcggcatc ctcatcgccc aggtgttcgg cctggactcc540 atcatgggca acaaggacct gtggcccctg ctgctgagca tcatcttcat cccggccctg 600 ctgcagtgca tcgtgctgcc cttctgcccc gagagtcccc gcttcctgct catcaaccgc aacgaggaga accgggccaa gagtgtgcta aagaagctgc gcgggacagc tgacgtgacc 720 catgacctgc aggagatga ggagagagt cggcagatga tgcgggaga gaggtcacc 780 atcctggagc tgttccgctc ccccgcctac cgccagccca tcctcatcgc tgtggtgctg 840 cagctgtccc agcagctgtc tggcatcaac gctgtcttct attack cagcatcttc gagaaggcgg gggtgcagca gcctgtgtat gccaccattg gctccggtat cgtcaacacg gccttcactg tcgtgtcgct gtttgtggtg gagcgagcag gccggcggac cctgcacctc 1020 ataggcctcg ctggcatggc gggttgtgcc atactcatga ccatcgcgct agcactgctg 1080 gagcagctac cctggatgtc ctatctgagc atcgtggcca tctttggctt tgtggccttc 1140 tttgaagtgg gtcctggccc catcccatgg ttcatcgtgg ctgaactctt cagccagggt 1200 ccacgtccag ctgccattgc cgttgcaggc ttctccaact ggacctcaaa tttcattgtg 1260 ggcatgtgct tccagtatgt ggagcaactg tgtggtccct acgtcttcat catcttcact 1320 gtgctcctgg ttctgttctt catcttcacc tacttcaaag ttcctgagac taaaggccgg 1380 accttcgatg agatcgcttc cggcttccgg caggggggag ccagccaaag tgacaagaca 1440 cccgaggagc tgttccatcc cctgggggct gattcccaag tg 1482 <210> 30 <211> 13 <212> DNA <213> Artificial Sequence <220> <223> Kozak sequence motif <400> 30 gccgccrcca ugg 13 <210> 31 <211> 10 <212> DNA <213> Artificial Sequence <220> <223> Kozak sequence motif <400> 31 gacaccaugg 10 <210> 32 <211> 141 <212> DNA <213> Adeno-associated virus <400> 32 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 60 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 120 actccatcac taggggttcc t 141 <210> 33 <211> 170 <212> DNA <213> Adeno-associated virus <400> 33 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 60 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 120 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta 170 <210> 34 <211> 141 <212> DNA <213> Adeno-associated viurs <400> 34 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg 60 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc 120 gagcgcgcag ctgcctgcag g 141 <210> 35 <211> 124 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - vector filler sequence <400> 35 gcggcaattc agtcgataac tataacggtc ctaaggtagc gatttaaata cgcgctctct 60 taaggtagcc ccgggacgcg tcaattgact acaaaccgag tatctgcaga gggccctgcg 120 tatg 124 <210> 36 <211> 84 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - vector filler sequence <400> 36 cttctgaggc ggaaagaacc agatcctctc ttaaggtagc atcgagattt aaattaggga 60 taacagggta atggcgcggg ccgc 84 <210> 37 <211> 63 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - vector filler sequence <400> 37 gttacccagg ctggagtgca gtggcacatt tctgctcact gcaacctcct cctccctggg 60 ttc 63 <210> 38 <211> 573 <212> DNA <213> Artificial Sequence <220> <223> Made in lab - CAG promoter in part Human betaherpesvirus 5 <400> 38 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 60 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 120 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 180 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 240 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga 300 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt 360 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggcgcg 420 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg 480 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc 540 ggccctataa aaagcgaagc gcgcggcggg cgg 573 <210> 39 <211> 253 <212> DNA <213> Homo sapiens <400> 39 gcccagcacc ccaaggcggc caacgccaaa actctccctc ctcctcttcc tcaatctcgc 60 tctcgctctt tttttttttc gcaaaaggag gggagagggg gtaaaaaaat gctgcactgt 120 gcggcgaagc cggtgagtga gcggcgcggg gccaatcagc gtgcgccgtt ccgaaagttg 180 ccttttatgg ctcgagcggc cgcggcggcg ccctataaaa cccagcggcg cgacgcgcca 240 ccaccgccga gtc 253 <210> 40 <211> 281 <212> DNA <213> Gallus gallus <400> 40 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc 60 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggg 120 ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg 180 cggcggcagc caatcagagc ggcgcgctcc gaaagttttcc ttttatggcg aggcggcggc 240 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg a 281 <210> 41 <211> 220 <212> DNA <213> Human betaherpesvirus 5 <400> 41 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 60 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 120 actttccaaa atgtcgtaat aaccccgcc cgttgacgca aatgggcggt aggcgtgtac 180 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg 220 <210> 42 <211> 583 <212> DNA <213> Human betaherpesvirus 5 <400> 42 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 60 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 120 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 180 atgggtggag tatttacggt aactgccca cttggcagta catcaagtgt atcatatgcc 240 aagtacgccc cctattgacg tcaatgacgg taatggccc gcctggcatt atgcccagta 300 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgcttattac 360 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggttg actcacgggg 420 atttccaagt ctccacccca ttgacgtca tgggagttg ttttggcacc aaaatcaacg 480 ggactttcca aaatgtcgta acaactccgc cccattgacg caatgggcg gtaggcgtgt 540 acggtgggag gtctatata gcagagctgg tttagtgaac cgt 583 <210> 43 <211> 508 <212> DNA <213> Human betaherpesviruses 5 <400> 43 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 60 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 120 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 180 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 240 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 300 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 360 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 420 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 480 acggtgggag gtctatataa gcagagct 508 <210> 44 <211> 573 <212> DNA <213> Artificial Sequence <220> <223> Made in lab - CAG promoter in part Human betaherpesvirus 5 <400> 44 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 60 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 120 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 180 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 240 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga 300 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt 360 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggcgcg 420 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg 480 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc 540 ggccctataa aaagcgaagc gcgcggcggg cgg 573 <210> 45 <211> 580 <212> DNA <213> Artificial Sequence <220> <223> Made in lab - CAG promoter in part Human betaherpesvirus 5 <400> 45 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 60 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 120 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 180 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 240 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 300 catgtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc 360 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg 420 ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg 480 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg 540 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg 580 <210> 46 <211> 455 <212> DNA <213> Homo sapiens <400> 46 caacctttgg agctaagcca gcaatggtag agggaagatt ctgcacgtcc cttccaggcg 60 gcctccccgt caccaccccc cccaacccgc cccgaccgga gctgagagta attcatacaa 120 aaggactcgc ccctgccttg gggaatccca gggaccgtcg ttaaactccc actaacgtag 180 aacccagaga tcgctgcgtt cccgccccct cacccgcccg ctctcgtcat cactgaggtg 240 gagaatagca tgcgtgaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag 300 tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg 360 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag 420 aaccgtatat aagtgcagta gtcgccgtga acgtt 455 <210> 47 <211> 401 <212> DNA <213> Homo sapiens <400> 47 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 60 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 120 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg 180 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc 240 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg 300 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg 360 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct c 401 <210> 48 <211> 448 <212> DNA <213> Homo sapiens <400> 48 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 60 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 120 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg 180 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc 240 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg 300 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg 360 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg 420 aggagtcgtg tcgtgcctga gagcgcag 448 <210> 49 <211> 422 <212> DNA <213> Homo sapiens <400> 49 ctgcagaggg ccctgcgtat gagtgcaagt gggttttagg accaggatga ggcggggtgg 60 gggtgcctac ctgacgaccg accccgaccc actggacaag cacccaaccc ccattcccca 120 aattgcgcat cccctatcag agagggggag gggaaacagg atgcggcgag gcgcgtgcgc 180 actgccagct tcagcaccgc ggacagtgcc ttcgcccccg cctggcggcg cgcgccaccg 240 ccgcctcagc actgaaggcg cgctgacgtc actcgccggt cccccgcaaa ctccccttcc 300 cggccacctt ggtcgcgtcc gcgccgccgc cggcccagcc ggaccgcacc acgcgaggcg 360 cgagataggg gggcacgggc gcgaccatct gcgctgcggc gccggcgact cagcgctgcc 420 tc 422 <210> 50 <211> 281 <212> DNA <213> Homo sapiens <400> 50 acttgtggac aaagtttgct ctattccacc tcctccaggc cctccttggg tccatcaccc 60 caggggtgct gggtccatcc cacccccagg cccacacagg cttgcagtat tgtgtgcggt 120 atggtcaggg cgtccgagag caggtttcgc agtggaaggc aggcaggtgt tggggaggca 180 gttaccgggg caacgggaac agggcgtttt ggaggtggtt gccatgggga cctggatgct 240 gacgaaggct cgcgaggctg tgagcagcca cagtgccctg c 281 <210> 51 <211> 851 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - eSYN promoter polynucleotide <400> 51 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 60 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 120 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 180 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc 240 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 300 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 360 tagtcatcgc tattaccatg gctgcagagg gccctgcgta tgagtgcaag tggttttag 420 gaccaggatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc cactgacaa 480 gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga ggggaaacag 540 gatgcggcga ggcgcgtcgc gactgccagc ttcagcaccg cggacagtgc cttcgccccc 600 gcctggcggc gcgcgccacc gccgcctcag cactgaggc gcgctgacgt cactcgccgg 660 tcccccgca actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc 720 cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccac tgcgctgcgg 780 cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct 840 gagagcgcag g 851 <210> 52 <211> 304 <212> DNA <213> Human betaherpesviruses 5 <400> 52 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 60 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 120 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 180 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 240 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 300 catg 304 <210> 53 <211> 953 <212> DNA <213> Homo sapiens <400> 53 cgcgtccgcc cgcgagcaca gagcctcgcc tttgccgatc cgccgcccgt ccacacccgc 60 cgccaggtaa gcccggccag ccgaccgggg catgcggccg cggcccttcg cccgtgcaga 120 gccgccgtct gggccgcagc ggggggcgca tggggcggaa ccggaccgcc gtggggggcg 180 cgggagaagc ccctgggcct ccggagatgg gggacacccc acgccagttc gcaggcgcga 240 ggccgcgctc gggcgggcgc gctccggggg tgccgctctc ggggcggggg caaccggcgg 300 ggtctttgtc tgagccgggc tcttgccaat ggggatcgca cggtgggcgc ggcgtagccc 360 ccgtcaggcc cggtgggggc tggggcgcca tgcgcgtgcg cgctggtcct ttgggcgcta 420 actgcgtgcg cgctgggaat tggcgctaat tgcgcgtgcg cgctgggact caatggcgct 480 aatcgcgcgt gcgttctggg gcccgggcgc ttgcgccact tcctgcccga gccgctggcg 540 cccgagggtg tggccgctgc gtgcgcgcgc gcgacccggt cgctgtttga accgggcgga 600 ggcggggctg gcgcccggtt gggagggggt tggggcctgg cttcctgccg cgcgccgcgg 660 ggacgcctcc gaccagtgtt tgccttttat ggtaataacg cggccggccc ggcttccttt 720 gtccccaatc tgggcgcgcg ccggcgcccc ctggcggcct aaggactcgg cgcgccggaa 780 gtggccaggg cggcagcggc tgctcttggc ggccccgagg tgactatagc cttcttttgt 840 gtcttgatag ttcgccagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc 900 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttc 953 <210> 54 <211> 1068 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - Chicken beta-actin exon / intron plus rabbit globin intron <400> 54 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc 60 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg 120 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc 180 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg 240 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc 300 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc 360 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc 420 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc 480 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc 540 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg 600 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccccggag cgccggcggc 660 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag 720 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcacccc 780 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc 840 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg 900 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc 960 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg 1020 ggcaacgtgc tggttatgt gctgtctcat cattttggca aagaattc 1068 <210> 55 <211> 126 <212> DNA <213> Homo sapiens <400> 55 agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagctg tgctcctggg 60 caccgcgcag tccgccccccg cggctcctgg ccagaccacc cctaggaccc cctgccccaa 120 gtcgca 126 <210> 56 <211> 121 <212> DNA <213> Human betaherpesvirus 5 <400> 56 tcagatcgcc tggagaggcc atccacgctg ttttgacctc catagtggac accgggaccg 60 atccagcctc cgcggccggg aacggtgcat tggaacgcgg attccccgtg ccaagagtga 120 c 121 <210> 57 <211> 512 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - adenovirus derived enhancer element <400> 57 ctcactctct tccgcatcgc tgtctgcgag ggccagctgt tgggctcgcg gttgaggaca 60 aactcttcgc ggtctttcca gtactcttgg atcggaaacc cgtcggcctc cgaacggtac 120 tccgccaccg agggacctga gcgagtccgc atcgaccgga tcggaaaacc tctcgagaaa 180 ggcgtctaac cagtcacagt cgcaaggtag gctgagcacc gtggcgggcg gcagcgggtg 240 gcggtcgggg ttgtttctgg cggaggtgct gctgatgatg taattaaagt aggcggtctt 300 gagacggcgg atggtcgagg tgaggtgtgg caggcttgag atccagctgt tggggtgagt 360 actccctctc aaaagcgggc attacttctg cgctaagatt gtcagtttcc aaaaacgagg 420 aggatttgat attcacctgg cccgatctgg ccatacactt gagtgacaat gacatccact 480 ttgcctttct ctccacaggt gtccactccc ag 512 <210> 58 <211> 956 <212> DNA <213> Homo sapiens <400> 58 ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg 60 cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctccag 120 tacgtgattc ttgatcccga gctggagcca ggggcgggcc ttgcgcttta ggagcccctt 180 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt 240 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat 300 gacgtgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caggatctgc 360 acactggtat ttcggttttt gggcccgcgg ccggcgacgg ggcccgtgcg tcccagcgca 420 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc 480 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg 540 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc 600 ctgctccagg gggctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac 660 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt 720 accgggcgcc gtccaggcac ctcgattagt tctggagctt ttggagtacg tcgtctttag 780 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag 840 ttaggccagc ttggcacttg atgtaattct ccttggaatt tggccttttt gagtttggat 900 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcag 956 <210> 59 <211> 939 <212> DNA <213> Homo sapiens <400> 59 gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg 60 ccttgaatta cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga 120 agtgggtggg agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt 180 gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt 240 ctcgctgctt tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt 300 tttttctggc aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt 360 tttggggccg cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg 420 ggcctgcgag cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct 480 ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg 540 tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca 600 aaatggagga cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg 660 gcctttccgt cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg 720 cacctcgatt agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt 780 tatgcgatgg agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac 840 ttgatgtaat tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag 900 cctcagacag tggttcaaag ttttttctt ccatttcag 939 <210> 60 <211> 83 <212> DNA <213> Homo sapiens <400> 60 tcagaagccc cgggctcgtc agtcaaaccg gttctctgtt tgcactcggc agcacgggca 60 ggcaagtggt ccctaggttc ggg 83 <210> 61 <211> 476 <212> DNA <213> Homo sapiens <400> 61 gtgagtctat gggacccttg atgttttctt tccccttcttt ttctatggtt aagttcatgt 60 cataggaagg ggagaagtaa cagggtacac atattgacca aatcagggta atttgcatt 120 tgtaatttta aaaaatgctt tcttcttta atatacttttt ttgtttatct tatttctaat 180 actttcccta atctctttct ttcagggcaa taatgataca atgtatcatg cctctttgca 240 cattctaaa gaataacagt gataatttct gggttaaggc aatagcaata tttctgcata 300 taatatttc tgcatataaa ttgtaactga tgtaagaggt ttcatattgc tatagcagc 360 tacaatccag ctaccattct gcttttattt tatggttggg ataaggctgg attattctga 420 gtccaagcta ggcccttttg ctaatcatgt tcatacctct tatcttcctc ccacag 476 <210> 62 <211> 589 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - mutated woodchuck hepatitis regulatory element <400> 62 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 60 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 120 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 180 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 240 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 300 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 360 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc 420 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 480 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 540 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgc 589 <210> 63 <211> 588 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - mutated woodchuck hepatitis regulatory element <400> 63 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 60 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 120 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 180 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 240 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 300 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 360 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 420 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 480 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 540 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgca 588 <210> 64 <211> 755 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - mutated woodchuck hepatitis regulatory element <400> 64 ttcctgttaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 60 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 120 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 180 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 240 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 300 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 360 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccgc 420 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 480 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 540 cgcctcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgccca 600 tgtatctttt tcacctgtgc cttgtttttg cctgtgttcc gcgtcctact tttcaagcct 660 ccaagctgtg ccttgggcgg ctttggggca tggacataga tccctataaa gaatttggtt 720 catcttatca gttgttgaat tttcttcctt tggac 755 <210> 65 <211> 12 <212> DNA <213> Artificial Sequence <220> <223> CAAX motif <400> 65 tgtgtgataa tg 12 <210> 66 <211> 810 <212> DNA <213> Homo sapiens <400> 66 ctgttctcat flawless caggttata flawless atgccacag atgttactta 60 gccttttaat atttctctaa ttgtgtat atgcaatgat agttctctga ttctgagat 120 tgagtttctc atgtgtaatg attatttaga gtttctctt catctgttca aatttttgtc 180 tagttttatt ttttactgat tgtaagact tctttttata atctgcatat tacaatctc 240 tttactgggg tgttgcaat attttctgtc attctatggc ctgactttc ttaatggttt 300 ttttaattta aaataagtc ttatattca tgcaatcta ttacaatct tttctttgtg 360 gttaggactt tgagtcataa gaaatttttc tctacactga agtcatg gcatgcttct 420 atattattt ctaaaagatt taaagttttg ccttctccat ttagacttat aattcactgg 480 aatttttg tgtgtatggt atgacatatg ggttcccttt tattttttac ataaataat 540 atttccctgt ttttctaaaa agaaaaaga tcatcatttt cccattgtaa atgccatat 600 tttttcata gtcacttac atatatcat gggtctgttt ctgagctcta ctctatttta 660 tcagcctcac tgtctatccc cacacatctc atgctttgct ctaatcttg atatttagtg 720 gaacattct tcccatttg ttctacaaga atattttgt tattgtcttt gggctttcta 780 tatacattt gaaatgaggt tgacaagtta 810 <210> 67 <211> 726 <212> DNA <213> Hepatitis B virus <400> 67 ataacaggcc tattgattgg aaagttgtc aacgaattgt gggtcttttg gggtttgctg 60 ccccttttac gcaatgtgga tatcctgctt taatgccttt atatgcatgt atacaagcaa 120 aacaggcttt tactttctcg ccaacttaca aggcctttct cagtaaacag tatatgaccc 180 tttaccccgt tgctcggcaa cggcctggtc tgtgccaagt gtttgctgac gcaaccccca 240 ctggttgggg cttggccata ggccatcagc gcatgcgtgg aacctttgtg tctcctctgc 300 cgatccatac tgcggaactc ctagccgctt gttttgctcg cagcaggtct ggagcaaacc 360 tcatcgggac cgacaattct gtcgtactct cccgcaagta tacatcgttt ccatggctgc 420 tagctgtgc tgccaactgg atcctgcgcg ggacgtcctt tgtttacgtc ccgtcggcgc 480 tgaatcccgc ggacgacccc tcccggggcc gcttggggct ctaccgcccg cttctccgtc 540 tgccgtaccg tccgaccacg gggcgcacct ctctttacgc ggactccccg tctgtgcctt 600 ctcatctgcc ggaccgtgtg cacttcgctt cacctctgca cgtcgcatgg aggccaccgt 660 gaacgcccac cggaacctgc ccaaggtctt gcataagagg actcttggac tttcagcaat 720 gtcatc 726 <210> 68 <211> 755 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - HepB derived enhancer element <400> 68 ttcctgtaaa caggcctatt gattggaaag tttgtcaacg aattgtgggt cttttggggt 60 ttgctgcccc ttttacgcaa tgtggatatc ctgctttaat gcctttatat gcatgtatac 120 aagcaaaaca ggcttttact ttctcgccaa cttacaaggc ctttctcagt aaacagtata 180 tgacccttta ccccgttgct cggcaacggc ctggtctgtg ccaagtgttt gctgacgcaa 240 cccccactgg ttggggcttg gccataggcc atcagcgcat gcgtggaacc tttgtgtctc 300 ctctgccgat ccatactgcg gaactcctag ccgcttgttt tgctcgcagc tggactggag 360 caaacctcat cgggaccgac aattctgtcg tactctcccg caagcactca ccgtttccgc 420 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 480 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 540 cgcctcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgccca 600 tgtatctttt tcacctgtgc cttgtttttg cctgtgttcc gcgtcctact tttcaagcct 660 ccaagctgtg ccttgggcgg ctttggggca tggacataga tccctataaa gaatttggtt 720 catcttatca gttgttgaat tttcttcctt tggac 755 <210> 69 <211> 94 <212> DNA <213> Homo sapiens <400> 69 gctggagcct cggtagccgt tcctcctgcc cgctgggcct cccaacgggc cctcctcccc 60 tccttgcacc ggcccttcct ggtctttgaa taaa 94 <210> 70 <211> 596 <212> DNA <213> Woodchuck hepatitis virus <400> 70 attcgagcat cttaccgcca tttattccca tatttgttct gtttttcttg atttgggtat 60 acatttaaat gttaataaaa caaaatggtg gggcaatcat ttacattttt agggatatgt 120 aattactagt tcaggtgtat tgccacaaga caaacatgtt aagaaacttt cccgttattt 180 acgctctgtt cctgttaatc aacctctgga ttacaaaatt tgtgaaagat tgactgatat 240 tcttaactat gttgctcctt ttacgctgtg tggatatgct gctttaatgc ctctgtatca 300 tgctattgct tcccgtacgg ctttcgtttt ctcctccttg tataaatcct ggttgctgtc 360 tctttatgag gagttgtggc ccgttgtccg tcaacgtggc gtggtgtgct ctgtgtttgc 420 tgacgcaacc cccactggct ggggcattgc caccacctgt caactccttt ctgggacttt 480 cgctttcccc ctcccgatcg ccacggcaga actcatcgcc gcctgccttg cccgctgctg 540 gacaggggct aggttgctgg gcactgataa ttccgtggtg ttgtcgggga agggcc 596 <210> 71 <211> 387 <212> DNA <213> Oryctolagus cuniculus <400> 71 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc 60 actcggaaga acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta 120 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg 180 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac 240 ttgaggttag attttttta tattttgttt tgtgttattt ttttctttaa catccctaaa 300 atttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat 360 agctgtccct cttctcttat ggagatc 387 <210> 72 <211> 251 <212> DNA <213> Taurus boss <400> 72 ttgccagcca tctgttgttt gccctcccc cgtgccttcc ttgaccctgg aaggtgccac 60 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 120 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aatacaatag 180 caggcatgct ggggatgcgg tgggctctat gggtacccag gtgctgaaga attgacccgg 240 ttcctcctgg g 251 <210> 73 <211> 251 <212> DNA <213> Taurus boss <400> 73 ttgccagcca tctgttgttt gccctcccc cgtgccttcc ttgaccctgg aaggtgccac 60 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 120 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 180 caggcatgct ggggatgcgg tgggctctat gggtacccag gtgctgaaga attgacccgg 240 ttcctcctgg g 251 <210> 74 <211> 225 <212> DNA <213> Taurus boss <400> 74 ctgtgccttc tagttgccag ccatctgttg tttgccccctc cccgtgcct tccttgaccc 60 tggaaggtgc cactcccact gtccttttcct aataaaatga ggaaattgca tcgcattgtc 120 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 180 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatgg 225 <210> 75 <211> 202 <212> DNA <213> Homo sapiens <400> 75 ctgcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctgccct ggaagttgcc 60 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag 120 gtgtccttct atatattat ggggtggagg ggggtggtat ggagcaaggg gcccaagttg 180 ggaagaaacc tgtaggccct gc 202 <210> 76 <211> 735 <212> PRT <213> Adeno-associated virus 2 <400> 76 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 Leu Lys Pro Gly Pro 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 Gly 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> 77 <211> 736 <212> PRT <213> Adeno-associated virus 9 <400> 77 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> 78 <211> 736 <212> PRT <213> Adeno-associated virus 6 <400> 78 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> 79 <211> 738 <212> PRT <213> Non-human primate Adeno-associated virus <400> 79 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> 80 <211> 738 <212> PRT <213> Adeno-associated virus 8 <400> 80 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> 81 <211> 738 <212> PRT <213> Non-human primate Adeno-associated virus <400> 81 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 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 Ser Pro Val 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 Asn 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 Asn 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> 82 <211> 743 <212> PRT <213> Artificial Sequence <220> <223> Synthetic construct - AAV9 variant <400> 82 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 Arg 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 Thr Leu Ala Val 580 585 590 Pro Phe Lys Ala Gln Ala Gln Thr Gly Trp Val Gln Asn Gln Gly Ile 595 600 605 Leu Pro Gly Met Val Trp Gln Asp Arg Asp Val Tyr Leu Gln Gly Pro 610 615 620 Ile Trp Ala Lys Ile Pro His Thr Asp Gly Asn Phe His Pro Ser Pro 625 630 635 640 Leu Met Gly Gly Phe Gly Met Lys His Pro Pro Pro Gln Ile Leu Ile 645 650 655 Lys Asn Thr Pro Val Pro Ala Asp Pro Pro Thr Ala Phe Asn Lys Asp 660 665 670 Lys Leu Asn Ser Phe Ile Thr Gln Tyr Ser Thr Gly Gln Val Ser Val 675 680 685 Glu Ile Glu Trp Glu Leu Gln Lys Glu Asn Ser Lys Arg Trp Asn Pro 690 695 700 Glu Ile Gln Tyr Thr Ser Asn Tyr Tyr Lys Ser Asn Asn Val Glu Phe 705 710 715 720 Ala Val Asn Thr Glu Gly Val Tyr Ser Glu Pro Arg Pro Ile Gly Thr 725 730 735 Arg Tyr Leu Thr Arg Asn Leu 740 <210> 83 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Peptide insert <400> 83 Thr Leu Ala Val Pro Phe Lys 1 5 <210> 84 <211> 7 <212> PRT <213> Artificial Sequence <220> <223> Peptide insert <400> 84 Lys Phe Pro Val Ala Leu Thr 1 5 <210> 85 <211> 940 <212> DNA <213> Homo sapiens <400> 85 tggagccgcc aaatattttg ggaaatagcg ggaatgttgg cgaactgggc aagtgcgttt 60 tctgattaag agcaaccaga ttcagctttt taaactacaa ttatactggc caaacaaaat 120 acccttatac aaaaaccaaa actactggca ggagtcgctg ccagcttgcg acccggcata 180 cttggctgag tatccgcttc tcccttgtgg ctccaaactg ctgcagattc tcggccactt 240 cagacgcgcg cgatggcgaa gagggtcctg cactttgacg cgcctggtga gggagcgctg 300 ctcttcgcag cgctcctggt gatgctcccc aaatttcggg gaccggcaag cgattaaatc 360 ttggagttgc tcagcgcccg ttaccgagta ctttttattt acaccagaaa caaagttgtt 420 gctctgggat gttctctcct gggcgacttg gggcccagcg cagtccagtt gtgtggggaa 480 atggggagat gtaaatgggc ttggggagct ggagatcgcc gccgggtacc cgggtgaggg 540 gcggggctgg ccgcacggga gagcccctcc tccgctccgg ccccgccccg catggccccg 600 cctccgcgct ctagagtttc ggcaccagct cccaccctgc actgagtccc gggaccccgg 660 gagagcggtc aatgtgtggt cgctgcgttt cctctgcctg cgccgggcat cacttgcgcg 720 ccgcagaaag tccgtctggc agcctggata tcctctccta ccggcacccg cagacgcccc 780 tgcagccgcg gtcggcgcc gggctccta gccctgtgcg ctcaactgtc ctgcgctgcg 840 gggtgccgg agttccacct ccgcgcctc ttctctagac aggcgctggg agaaagaacc 900 ggctcccgag ttctgggcat ttcgcccggc tcgaggtgca 940 <210> 86 <211> 1142 <212> DNA <213> Mus musculus <400> 86 aagcttccga ccgttagtca gagaactgta agtgctcaga gcctggctga caatgatctg 60 gaatgaacca gataacaaca tataaaatc tcagtaaaat aatttaacag ttagcttgga 120 agctggtcag ctctggggaa atcagggtaa attgtgctgt catgaactgt cccacactga 180 catcggccaa agtgaatatg aactttggta gatccaatgc ctgttctatt tatttttcca 240 gtgaaaagta ttttgataga gcttttcatt ttgtaaatac actgagttaa ccaaaatatc 300 atggattcc gtttgttctt aagacatgca actcgtctac ggctatacca ctctgaacgc 360 gccgatctc ggaagacatg caactcaaat gtaaatacag tagaatatta cttaggtaga 420 aactcctggt gattttaaaa gattggaaaa gaatatgagg aagagttgaa taatgcaaat 480 tctagtgtgt gtgctaccga agtgaacact taatgcacag tctacagact aggacatttt 540 atcgtgtgtt gtaaaattgg gtagaaactt gtgtttgtga aaactgagca ttaaaacctt 600 acagagaccg tttcttgttt acttttgaaa aaaaaaagag tcacgtgagc ctcattttgt 660 atttgtgtgt gtgtgtgtgt gtgtgtctcc cctcctccca gcgtgtgtgt gctgggagga 720 ggggagaccc cagaacaatg tcctgcctcc aaaccttctc aataggcgga agccactggc 780 ttcctccctt tcctgtctcc cgtgctccag caatgcagat ggaagggacc gaagggatgg 840 gagagagagc ccaaccatcc ccagatctgt ccttgtcaca acctgcctcc cacctctaat 900 gccccccctt ccagagactt ccaggccaca cccatcccgg gcttgtgggg gctggacacg 960 ggaggactac aggcgacaac tcttcccacc ctctctccct gccacccctc ctaccctaac 1020 catcatttcc tcttcctccc cagcaccgag gtgcactgag ctggacactca tgaacactca 1080 gacccacagc aactgacccc gggcccagct ggccttggct ggcccaggggc agcttccaga 1140 gt 1142 <210> 87 <211> 2079 <212> DNA <213> Homo sapiens <400> 87 gctggagtgc agtggcacga tctcggctca ctgcaacctc tgcctcccag gttcaaacaa 60 ttctcctgcc tcagcctcca gagtagctgg ggttacaggt gcacgccagc aagcacagct 120 aaattttgta tttttagtag agatggggtt ttgccatgtt ggccaggctg gtctcaaact 180 cctgacctca ggtgatccac tcccaaagtg ctgggattat aggcgtgagc cactgtgcca 240 ggcccactgt ttttgttttt ttttttcgtg atgacaaatt taaagtcatc tcataggaat 300 agaaaatagc tttttagtag aagctcttgg aatttaaatt gagactgaat ggaaagatga 360 aagaaaataa acttattaac atttaatgag aaccttcaaa gaactaggca tagtaccaaa 420 tggttttata tttttaaacc tcatttattc ctctcaaaac acctgggaag gagatatttt 480 tgccatttca cagctgttga aactgaggct caaaaagact aagtaacttt tctcagctac 540 acatgtggct gagccagtat ttgaacccag ttctgtttgc agacagaacc tgggcttttt 600 cacacctgca aactggaaac attaattggt tcttaagatc atcatcgatg tgataaaaacc 660 tgggacagaa attagtcaag actagctgca tctgcctttt cctctggtgg gtaggaaaag 720 gaggagtata atgatttcct caggcatgaa ggtcgatgat gagcaaagtg tatactctct 780 aatctaatgt cataattcat attgtggagt aattatctgg ataagtgtag ggtctctgac 840 ctcattctag atattgtaca ttccatggct atttcattt tggtccatga actctctttg 900 ctctcatgag caccattttt atcccaatct aatcctgtat gtttgtgttt ttacacagat 960 tagttttaa atgttatata taatttgctt ctgaaacacc attgctcaat gactaccaaa 1020 tctttctcat taccaaaatc cttctatgcc aacttcttca agaaatttga tcacctttag 1080 atgaattgtt aatgaaaatt aaagctatag ccggcaacat gggtatcttt gggctaatgg 1140 ccaaccaaca ggccatctgt gtgaaagaaa acaggctaac aattttggac tctggtctct 1200 tggggctaca ttgagcattg acctcaccgg tgctcactga aattaattgc ttttcaggtt 1260 gtattttctc atcacggaaa ccttcttctc ccaattcaaa ccatgtgggt taaaatgaga 1320 aaacaaaagc caaaacggct tcccacaccc aaaagctcct tctgtcagag atcccagtag 1380 ccccgggaga gctgttagaa gtctgagaag gattggtcat catcgcatac catacatagg 1440 tggagggctt gttattctca gtttcccgcc tatgagagga tacccctatt gtttctgaaa 1500 atgctgaccg ggacccacac ttccaacaaa aattcctctg cccctacagc agcagcaaaa 1560 gcagcagcag aagcaacagc aacagataag tgttttgatg aattgcgaga tggatagggc 1620 ttgagtgccc ccagccctgc tgataccaaa tgcctttaag atacagcctt tcccatccta 1680 atctacaaag gaaacaggaa aaaggaactt aaactccct gtgctcagac agaaatgaga 1740 ctgttacagc ctgcttctgt gctgttcctt cttgcctcta acttgtaaac aagacgtagt 1800 aggacgatgc taatggaaag tcacaaaccg ctgggttttt gaaaggatcc ttgggacctc 1860 atgcacattt gtggaaactg gatggagaga tttggggaag catggactct ttagccagct 1920 tagttctctg tggagtcagc ttgctccttt ctggtaaggt ttggctttat tttttttaat 1980 ttagtatttt aaaaaacaga gttagtgatt tctgggtgct ctccccaaat ctcatcagtg 2040 ctgatgaaca aggggtggct gtagcaaagg caccatttc 2079 <210> 88 <211> 1559 <212> DNA <213> Homo sapiens <400> 88 catccatgcc catggcctca gatgccagcc ataagctgtt gggttccaaa cctcgactcc 60 aggctggact cacccctgtc tcccccacca gcctgacacc tccacctggg tatctaacga 120 gcatctcaaa ctcaacctgc ctgagacaga ggaatcacta tcccctcctc ctccaaaaat 180 atccttccat cacactcccc atcttgtgct ctgatttact aaacggccct gggccctctc 240 tttctcaggg tctctgcttg cccagctata tataaaaca agtttgggac ttcccaacca 300 ttcacccatg gaaaaacaga agcaactctt caaaggacag attcccagga tctgccctgg 360 gagattccaa atcagttgat ctggggtgag cccagtcctc tgtagttttt agaagctcct 420 cctatgtctc tcctggtcag cagaatctttg gccctccct tccccccagc ctctttggttc 480 ttctgggctc tgatccagcc tcagcgtcac tgtcttccac gcccctcttt gattctcgtt 540 tatgtcaaaa gccttgtgag gatgaggctg tgattatccc cattttacag atgaggaaac 600 tgtggctcca ggatgacaca actggccaga ggtcacatca gaagcagagc tgggtcactt 660 gactccaccc aatatcccta aatgcaaaca tcccctacag accgaggctg gcaccttaga 720 gctggagtcc atgcccgctc tgaccaggag aagccaacct ggtcctccag agccaagagc 780 ttctgtccct ttcccatctc ctgaagcctc cctgtcacct ttaaagtcca ttcccacaaa 840 gacatcatgg gatcaccaca gaaaatcaag ctctggggct aggctgaccc cagctagatt 900 tttggctctt ttatacccca gctgggtgga caagcacctt aaacccgctg agcctcagct 960 tcccgggcta taaaatgggg gtgatgacac ctgcctgtag cattccaagg agggttaaat 1020 gtgatgctgc agccaagggt ccccacagcc aggctctttg caggtgctgg gttcagagtc 1080 ccagagctga ggccgggagt aggggttcaa gtggggtgcc ccaggcaggg tccagtgcca 1140 gccctctgtg gagacagcca tccggggccg aggcagccgc ccaccgcagg gcctgcctat 1200 ctgcagccag cccagccctc acaaaggaac aataacagga aaccatccca gggggaagtg 1260 ggccagggcc agctggaaaa cctgaagggg aggcagccag gcctccctcg ccagcggggt 1320 gtggctcccc tccaaagacg gtcggctgac aggctccaca gagctccact cacgctcagc 1380 cctggacgga caggcagtcc aacggaacag aaacatccct cagcccacag gcacggtgag 1440 tgggggctcc cacactcccc tccaccccaa acccgccacc ctgcgcccaa gatgggaggg 1500 tcctcagctt ccccatctgt agaatgggca tcgtcccact cccatgacag agaggctcc 1559 <210> 89 <211> 399 <212> DNA <213> Homo sapiens <400> 89 gtctcccagg catgactcca acaatgcatc ccatgggatt tggggttccc cagatctggg 60 gcttgtaggc ctgactctcc cctgtgcaca cgtctcatac acgcatgcgt gcacccattg 120 cctgccccgc cccttgcaca gggagtcagc agggaggact gggttatgcc ctgcttatca 180 gcagcttccc agcttcctct gcctggattc ttagaggcct ggggtcctag aacgagctgg 240 tgcacgtggc ttcccaaaga tctctcagat aatgagagga aatgcagtca tcagtttgca 300 gaaggctagg gattctgggc catagctcag acctgcgccc accatctccc tccaggcagc 360 ccttggctgg tccctgcgag cccgtggaga ctgccagtc 399 <210> 90 <211> 735 <212> DNA <213> Homo sapiens <400> 90 atctttagcc gatccattca accctggcca ggatccaaat ggactgtttt tgtcagggcc 60 aggaccggat ccttcatacc tggggtgcat aggaagtgtt agtactcccc ttcctccaaa 120 cacagcagca aaattggctc aggttgaggt gttttctca acttccctgg agtccagccc 180 tggaagctgg atcaggaagc tgtgttgttc tactgtgatt ccccctggcc tgtatcagct 240 tgccctgaaa caaccagcat tcctggttat cccacacagg tggggcactc taggaagacc 300 agggatcaag tgtgggggtg tagggatagg gggtgtttgg ggagggcaag gcagttaatt 360 aaggcagctg ccaggaggtc tccctccaaa ctctacaaag ctttatcagc ttggaggtac 420 ttctaatacc atttcctttc attgtttcct tttggtaatt aaaaggaggc caatcccctg 480 ttgtggcagc tcacagctat tgtggtggga aagggagggt ggttggtgga tgtcacagct 540 tgggcttat ctcccccagc agtggggact ccacagcccc tgggctacat aacagcaaga 600 cagtccggag ctgtagcaga cctgattgag cctttgcagc agctgagagc atggcctagg 660 gtgggcggca ccattgtcca gcagctgagt ttcccaggga ccttggagat agccgcagcc 720 ctcatttgca gggga 735 <210> 91 <211> 1132 <212> DNA <213> Homo sapiens <400> 91 tggcttccgg agggtggcct gggggctggg gtgccaggga caccatcgcc actggtggga 60 gggcagggca cagcccctcc gtgtcccttt gtctctcctg tctgaaggcc agagcaggct 120 gctaggcctg gggccaccac tgcccctggg tgctacaccc agtgtgctgg gtcactggga 180 acttcctgaa gtggtgtcac ctgaactggg cccccaagga tggggtgcgg gcagtaccgc 240 aggaagagga gcagcccctg tgaagattga gaggtctggg aagcccctgc ggcttgggag 300 agtgggggtc gccaggcagg gggaaagccc ctgtgccacc gctttttgcc agagactcag 360 gctccagaga ggcagtgagt ggcatggggg gtgaggctgg ggccctgggc ctgacctcca 420 cacgcctgcc tggcctctct gtttgccatg ggatgagaga gacagtgctg ggactcagag 480 cggggctgga gagtgagagt gcgagaaagg gcctgggtgg ggcttggacc ccggggcggg 540 ctttctggag agccccccta cgagggcctc tacggcggtg acggggtggg gggcttctgc 600 aaaccttggt cagggaagtg gagctggctc gagtggaaga gaccacccgg ctcagtcggg 660 gatgtgggag tggactgggt ggtgcagact gggggtcgag cgccttctga agtgacgggg 720 ccgggacgcg cagggaggcg gcccaagaag cgcgccctag gccagcccag aatgcgctcg 780 gccgcgacta ggacaacggc gggtggggct gggggcggct gccgggcggg gagcggtccc 840 gcgccctcag ctacccctca agagccgttg tttccctaac ttcagctgcc agaggctctg 900 tgattggctg cggcacgatg acccgcgcac ggattggctg cttcgggccg gggggccggg 960 cccgggggac agaatccgcc cccgaacctt caaagagggt accccccggc aggagctggc 1020 agacccagga ggtgcgacag acccgcgggg caaacggact ggggccaaga gccgggagcg 1080 cgggcgcaaa ggcaccaggg cccgcccagg gcgccgcgca gcacggcctt gg 1132 <210> 92 <211> 888 <212> DNA <213> Homo sapiens <400> 92 cgccttgctg tgccactttg ggacttccct ccctagcctg agcttcagtt ttcctgcctg 60 ttaggcagcc ccatgtcaac tgcacttagt aggccgggtt tgatgcccga caagacgtga 120 agtggtggag gtgggcagga tcccagcgct accatcttct tgaaccagtg atctcaacac 180 atcggatttc tgtttcctca tctgcaaaat gggatcagtg agctcaggtg ggtcacaaat 240 tctacaggaa ctactttagc caagcccggc cccctgaaag ttcccctcgg tgggctgtta 300 gggtgattgt tttcatctgt ggggctccct gatgcgtccc acccaccagc cttggagagg 360 gtgggatggg agggtggggt gcttggggag acaagcctag agcctgggcc ctcccacccc 420 actgcctccc cccatcccag ggccccccac ccagtgacaa agcccgtggc acttcctcta 480 cccggttggc aggcggcctg gcccagcccc ttctctaagg aagcgcattt cctgcctccc 540 tgggccggcc gggctggatg agccgggagc tccctgctgc cggtcatacc acagccttca 600 tctgcgccct ggggccagga ctgctgctgt cactgccatc cattggagcc cagcaccccc 660 tccccgccca tccttcggac agcaactcca gcccagcccc gcgtccctgt gtccacttct 720 cctgacccct cggccgccac cccagaaggc tggagcaggg acgccgtcgc tccggccgcc 780 tgctcccctc gggtccccgt gcgagcccac gccggccccg gtgccccccc gcagccctgc 840 cactggacac aggataaggc ccagcgcaca ggcccccacg tggacacc <210> 93 <211> 1658 <212> DNA <213> Homo sapiens <400> 93 gcccaggctg gagtgcagtg gcacagtcac aactcactgc agcctcaaac tcctgggctc aaaacgatcc acagtctcct gagtagctgg gactacagga gcttgttacc acacccagct ccagtttata aattcatctc cagtttata aggaggaac cgaggtactg aggaggatta aaaccttcct gcagacactt gtccagcaag tggccactcc aggatttgga ccaaggtgat 240 gtgtcttcag gctgtgtctc tgccactgtg ccacgctgct gggtggtagg cagcagtggg 300 tgggtgcctg cagtggtctg taaagaccac ctgagatgtc cttcctcctc tgttccaccc 360 420. tgtccaggtc caagaagaca gtctatgaag agagagcagg tgtgactctc tcagtgtgct cctctgtgag aagcaggctg acatcccaaa gggaagggcg gataacagag acagtgcaag 480 cggaggagat gagggtgcct caaagccggg aggctgggtg atgcaggagc ctgcgtgtcc 540 cgaggggggt gctgggccca gtgtgagtac gtgtgactgt gactgagaca gtgtgactgc 600 tgaaggcagg gacacagcag ctccctgact gggggcagaa ggcgttaact gtgtgaaggc 660 tggttgtggg tgggtgggct ctgggcctcg aacccggggg ctgagggaga tagtaaacag 720 cagggtgact gacgggaaga tcatgttggt agccctgcga agatgctgca gggctgtggg 780 ggtttgtgtg actttgcagt tcaacaaatt caaattcagc caacgctggc agggcctgtt 840 gtgccaggca accagctagg aggaggagac tcggacccag cttgcagctg aagggcgctg 900 gctgccgggt tctgtgggtt caccttgcgg tgtcttccct tgctaacact gagtccttac 960 aatagcccca tctccaggtt gaggctagat ggaggggaca gagggaagtg acttgcccaa 1020 ggtgacccaa gctcccgagt gccagggcag gatctgaatt caggctctca gactgcagag 1080 cctgagtccc tccctgccat gcctgtgcca gggtggaaat gtctggtcct ggaggggagc 1140 gtggactcct ggccttggct ctggagacat ccccctagac cacgtgggct ctaacctgt 1200 ccatggtcac tgtgctgagg ggcgggacgg tgggtcaccc ctagttcttt tttccccagg 1260 gccagattca tggactgaag ggttgctcgg ctctcagaga ccccctaagc gccccgccct 1320 ggccccaagc cctcccccag ctcccgcgtc cccccctc tggcgctgac tccgggccag 1380 aagaggaaag gctgtctcca cccacctctc gcactctccc ttctccttta taaaggccgg 1440 aacagctgaa agggtggcaa cttctcctcc tgcagccggg agcggcctgc ctgcctccct 1500 gcgcacccgc agcctccccc gctgcccc tagggctccc ctccggccgc cagcgcccat 1560 ttttcattcc ctagatagag atactttgcg cgcacacaca tacatacgcg cgcaaaaagg 1620 aaaaaaaaa aaaaaagcc accctccagc ctcgctgc 1658 <210> 94 <211> 1455 <212> DNA <213> Homo sapiens <400> 94 acatccaatg cccgctctgc ctcatcttct atgggaaaca agaattttag aggtcaggta 60 gcctaacacc atcaattctc aaaagaggaa gctgaggcca agagaagtc tgtgaatttc 120 ttacagctca tttgtgacag accaagaatt acccacttta ctgggttgtt atttactaag 180 tgacagtgag tctatatctc ttttgacaag tgaggtgggg gcatggaatt cggcatgtgg 240 ttggtgtaag aactcccctc tctcctcttt aaccttactt aataagaccc tggcacagtt 300 gatattttaa gagggctact ctgttttccc agagggacct aggcacggta accctcttag 360 catgcagacc ttgtttcctg aggggtaatg tttcccttcc ctgtgacttg tttcttgggg 420 gctgtgttct gattttcctg ctgagccact tgttgccttg ggctggctgc cgcgcttggc 480 agtttttagt gagggctctg atagatgcca ggaggtgagg ggaagggctc tgggtggact 540 ccgtcattgg acaagcagac ttagtgatgg atgagccttc ccctgaggaa gttttggatc 600 agaagtccaa ctgataagtt tttccagaat tgagtaaccc agaagcagtg ccgaaaggat 660 cttacctctc ttgtggcttt ttgtattgat tttaaaagaa attctcagag gcagttccac 720 attgtactgg aagcacagct atatccacaa taggcttaga tatatgtaac atgaattgct 780 ttagaaataa catttgagga gaggggtgag aggaaggaag agagggtctt aaaaatagc 840 cctatcaaaa tatttcttt cttctaagta ttgaaaagac acaatataac cctttcttct 900 ttcaaatgat ctcatagcta tttgttgagg ggaaatacca aatgtttatt atttttttg 960 aagaagcttc ttcggtcctg atgattcatg ttgatatcat tttcctcctg actacagagg 1020 ctctgagaca aagctacacc tcaagtgata tgccagggtc agaacaattc ccgtcctgaa 1080 ggagggtgtg caaccttctt tatccctcct tcacagacgt ccttgagccc ttgagacgga 1140 tgtgagtgag tttttcagtc ctcatgcaaa acaaccatct aaacataaca gatgacatca 1200 gcttgggct ttcaattcct ggatggcagc agcgtgttaa tccagccttc atcctggatt 1260 tcataaacca aaacaagaga gcctggcagg aggacagcgc tgctgctggg ttgaggaaat 1320 tgatgacggg aaagcatgcg ggcaacccag tgtataaaac tcataaacgt gtaggcagag 1380 gctcagctac cagtttggac ggctgcttcc caccagcaaa gaccacgact ggagagccga 1440 gccggaggca gctgg 1455 <210> 95 <211> 1389 <212> DNA <213> Homo sapiens <400> 95 tggcacacac gcaccctgtc caatgtatct tttgtgtaaa tctggactta acacttcaag 60 caaactgcct ggcttgctga aaggtggaga cacctttcga ttcagtcttt tatatgtgt 120 tgagtgccac ctatgtgcag agcaagatat tggggacttt ggagagatcc agaagagtga 180 gaagacagta tcctacctta gggggttccc agtccaatga gggaagcagc cccatgcctt 240 gggagctccc aagctataga agcagctaac aatcgagtct ggaaaggcaa acaacttcag 300 gacccgcttc taagcggaa tcgcaagtac acgcaaaatg aatccagcct tgactgtgtg 360 gagttgggta aaccacctgc ctcttacgtt gatggggaac tagaatgagg acagctccag 420 ggaacaagaa agggtagacc ataggagctg tcccatgtcc caacagtggg gaggagctga 480 tgggcggccc ctgctggatt agtgttatcc tgagaaggct tctggatgcg atgggatttg 540 aggtgctgct gcaaagaatg aattgctcac ggaagggtgg ggtgggggca ttccaggtag 600 agggtgcctc ctgggggatg cagggaacat gaggggcctg ggcaattaat caagccttgg 660 gcacaagcct aggcagtcac ccccaattca aagccagttg aaaatgcaga ggagagagga 720 gggccagtgt ttggttgtct tgaccaaacc cttgaagctg gccagcggca agggcaagga 780 ccagggtcag aggtagaggg cgtgagtgaa ggcaacccag actgagtcct tccctaagcg 840 cccaggtttc ctgacagctg ttaaggaagc aaggtgagaa agggttaagt gtgcccctcc 900 accgccccaa atgcttcctg tgtttgaaat ccttcaggtc tctgcaaacc ctctggcccc 960 cggccaggcg ggcattgtcc ggggagcggt tgtaggttgt cagagaggcc gcgcagcctt 1020 tgttgtgggg ccacctcggg gttccctctc gcgctcacgc tcgggctggg gctgcagagt 1080 gcgtgcctgg aggggggcgg tgcgggaggc tcgctccctc tccctcttcc tgccccccct 1140 ctagccctcc cgatgaccac atgaccaagt gggctcgcgg ccaagccaca agctacaaaa 1200 tgcagcccct ggagtgagcg gggagcattc tctctggcag ccggggtcac gggcagttgc 1260 agccgcggcc gagcagccag ccgctaagaa agagctcgcc gctgccgctc ccggagccgc 1320 cgaggccagc ttcgcggcgc tgccccgcgg cgggagagga ggctgcagaa gagcggaggc 1380 ggccagcgg 1389 <210> 96 <211> 4258 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - full polynucleotide sequence of vector genome <400> 96 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 120 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtact ctggagacgc 180 gttacataag ctcctcccag cctcaggccc aggaatggga atctctgtgg gtcacacatc 240 agtagggagg tctttcccga tccttttcta tgctactcca ggagtcaaag cgtctcctgg 300 gacttttcag ggcgcttcag aagagccctg ggcctaaacc agctcaacca agctgcaggg 360 acccagcctc ctgagaaaag tgaatgtgag cccggtgcat tcagaggaga atgaagcctt 420 cacccagaac acactctggg aagatgtccc aggcccaggg ggagggtttg tactaccaga 480 cctaagtcac ctaaactgac accaagtctc atccatccca accattccat tccgggtcag 540 aggggtcatc gatttaacca gcaaggctgc ccatccaacg gttgctccct ctgctccctg 600 gaagggcctc ctcgtgggcg ttctgtacct acaggtcttg ttccgttctg ggaactgcca 660 gtggtggcaa gaggtggagc aacgggtgcc agggcaggga gaggtgagtc tgggagggaa 720 gcagaggcaa gatccatggg gctttagaga ctttgccaaa gcagtgcgac tgctcccagg 780 ttgttgtcag ccgtcaagag tgagtgcacc tccctgggca gacttctgct gccccagtgc 840 ccaggaatag gcaggggttt gccgcaaaat gaatgacacc tggcagacaa taagctgaag 900 ctttcattag cagcttaagc tgaggactat ctatgcaacc gatactccct gtgtgctccc 960 cgggactgct taatgtgagc ccttgtggag cgattggcac caagaaagca aggactaagt 1020 cagaagttca agtcccagcc ttgccacagc ctcagggtgc cctcgagcac agcaagcctc 1080 agttttccca tctgtacaat gagagaggta cacaaggtag actcgaaggc tctttgttgc 1140 cagggccctg tgttcctttg agtgtatgtg cttctcaggc ccacagaggt cctttgtgtt 1200 tcgtatgtga actgctctct aggaaaccca tgtaactgtc tgtgtcctgg ggcacataca 1260 tgaggactca tgtgggccgt attgtgtgtt tgtgccgggg ggaggggaga ccccagaaca 1320 atgtccccca ccccaccccc ctcctcaata ggcggaagcc actggcttcc tccctttcct 1380 gcctcctgcc tcctttgtgc cagcaagact gagtactgga gagagacagg ggatgggaaa 1440 aatcagtcca gctgtcccca ggtctgccct taccataacc ttccccccac ctcaagtgac 1500 tcctcccagg ccacacccat ccccagcctt gtgggggcca gattgggggg cctagaggct 1560 caaaggcaga atgagtcctc ccacccccta ccctgccacc cctcccaccc aagccacctc 1620 atttcctctt cctccccagc accgacccac actgaccaac acaggctgag cagtcaggcc 1680 cacagcatct gaccccaggc ccagctcgtc ctggctggcc tgggtcggcc tctggagtgc 1740 caccatggag cccagcagca agaagctgac gggtcgcctc atgctggccg tgggaggagc 1800 agtgcttggc tccctgcagt ttggctacaa cactggagtc atcaatgccc cccagaaggt 1860 gatcgaggag ttctacaacc agacatgggt ccaccgctat ggggagagca tcctgcccac 1920 cacgctcacc acgctctggt ccctctcagt ggccatcttt tctgttgggg gcatgattgg 1980 ctccttctct gtgggccttt tcgttaaccg ctttggccgg cggaattcaa tgctgatgat 2040 gaacctgctg gccttcgtgt ccgccgtgct catgggcttc tcgaaactgg gcaagtcctt 2100 tgagatgctg atcctgggcc gcttcatcat cggtgtgtac tgcggcctga ccacaggctt 2160 cgtgcccatg tatgtgggtg aagtgtcacc cacagccctt cgtggggccc tgggcaccct 2220 gcaccagctg ggcatcgtcg tcggcatcct catcgcccag gtgttcggcc tggactccat 2280 catgggcaac aaggacctgt ggcccctgct gctgagcatc atcttcatcc cggccctgct 2340 gcagtgcatc gtgctgccct tctgccccga gagtccccgc ttcctgctca tcaaccgcaa 2400 cgaggagaac cgggccaaga gtgtgctaaa gaagctgcgc gggacagctg acgtgaccca 2460 tgacctgcag gagatgaagg aagagagtcg gcagatgatg cgggagaaga aggtcaccat 2520 cctggagctg ttccgctccc ccgcctaccg ccagcccatc ctcatcgctg tggtgctgca 2580 gctgtcccag cagctgtctg gcatcaacgc tgtcttctat tactccacga gcatcttcga 2640 gaaggcgggg gtgcagcagc ctgtgtatgc caccattggc tccggtatcg tcaacacggc 2700 cttcactgtc gtgtcgctgt ttgtggtgga gcgagcaggc cggcggaccc tgcacctcat 2760 aggcctcgct ggcatggcgg gttgtgccat actcatgacc atcgcgctag cactgctgga 2820 gcagctaccc tggatgtcct atctgagcat cgtggccatc tttggctttg tggccttctt 2880 tgaagtgggt cctggcccca tcccatggtt catcgtggct gaactcttca gccagggtcc 2940 acgtccagct gccattgccg ttgcaggctt ctccaactgg acctcaaatt tcattgtggg 3000 catgtgcttc cagtatgtgg agcaactgtg tggtccctac gtcttcatca tcttcactgt 3060 gctcctggtt ctgttcttca tcttcaccta cttcaaagtt cctgagacta aaggccggac 3120 cttcgatgag atcgcttccg gcttccggca ggggggagcc agccaaagtg acaagacacc 3180 cgaggagctg ttccatcccc tgggggctga ttcccaagtg tgataatgga tcaacctctg 3240 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta 3300 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt 3360 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc 3420 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt 3480 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg 3540 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac 3600 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc 3660 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac 3720 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct 3780 cagacgagtc ggatctccct ttgggccgcc tccccgcatc attgcctgcc cgggtggcat 3840 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc 3900 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat 3960 attatggggt ggaggggggt ggtatggagc aaggggccca agttgggaag aaacctgtag 4020 ggcctgcgtt acccaggctg gagtgcagtg gcacatttct gctcactgca acctcctcct 4080 ccctgggttc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct 4140 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc 4200 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgc 4258 <210> 97 <211> 3922 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - part of expression cassette <400> 97 ctctggagac gcgttacata agctcctccc agcctcaggc ccaggaatgg gaatctctgt 60 gggtcacaca tcagtaggga ggtctttccc gatccttttc tatgctactc caggagtcaa 120 agcgtctcct gggacttttc agggcgcttc agaagagccc tgggcctaaa ccagctcaac 180 caagctgcag ggacccagcc tcctgagaaa agtgaatgtg agcccggtgc attcagagga 240 gaatgaagcc ttcacccaga acacactctg ggaagatgtc ccaggcccag ggggagggtt 300 tgtactacca gacctaagtc acctaaactg acaccaagtc tcatccatcc caaccattcc 360 attccgggtc agaggggtca tcgatttaac cagcaaggct gcccatccaa cggttgctcc 420 ctctgctccc tggaagggcc tcctcgtggg cgttctgtac ctacaggtct tgttccgttc 480 tgggaactgc cagtggtggc aagaggtgga gcaacgggtg ccagggcagg gagaggtgag 540 tctgggagg aagcagaggc aagatccatg gggctttaga gactttgcca aagcagtgcg 600 actgctccca ggttgttgtc agccgtcaag agtgagtgca cctccctggg cagacttctg 660 ctgccccagt gcccaggaat aggcaggggt ttgccgcaaa atgaatgaca cctggcagac 720 aataagctga agctttcatt agcagcttaa gctgaggact atctatgcaa ccgatactcc 780 ctgtgtgctc cccgggactg cttaatgtga gcccttgtgg agcgattggc accaagaaag 840 caaggactaa gtcagaagtt caagtcccag ccttgccaca gcctcagggt gccctcgagc 900 acagcaagcc tcagttttcc catctgtaca atgagagagg tacaaaggt agactcgaag 960 gctctttgtt gccagggccc tgtgttcctt tgagtgtatg tgcttctcag gcccacagag 1020 gtcctttgtg tttcgtatgt gaactgctct ctaggaaacc catgtaactg tctgtgtcct 1080 ggggcacata catgaggact catgtgggcc gtattgtgtg tttgtgccgg ggggagggga 1140 gaccccagaa caatgtcccc caccccaccc ccctcctcaa taggcggaag ccactggctt 1200 cctccctttc ctgcctcctg cctcctttgt gccagcaaga ctgagtactg gagagagaca 1260 ggggatggga aaaatcagtc cagctgtccc caggtctgcc cttaccataa ccttcccccc 1320 acctcaagtg actcctccca ggccacaccc atccccagcc ttgtgggggc cagattgggg 1380 ggcctagagg ctcaaaggca gaatgagtcc tcccaccccc taccctgcca cccctcccac 1440 ccaagccacc tcatttcctc ttcctcccca gcaccgaccc acactgacca acacaggctg 1500 agcagtcagg cccacagcat ctgaccccag gcccagctcg tcctggctgg cctgggtcgg 1560 cctctggagt gccaccatgg agcccagcag caagaagctg acgggtcgcc tcatgctggc 1620 cgtgggagga gcagtgcttg gctccctgca gtttggctac aacactggag tcatcaatgc 1680 cccccagaag gtgatcgagg agttctacaa ccagacatgg gtccaccgct atggggagag 1740 catcctgccc accacgctca ccacgctctg gtccctctca gtggccatct tttctgttgg 1800 gggcatgatt ggctccttct ctgtgggcct tttcgttaac cgctttggcc ggcggaattc 1860 aatgctgatg atgaacctgc tggccttcgt gtccgccgtg ctcatgggct tctcgaaact gggcaagtcc tttgagatgc tgatcctggg ccgcttcatc atcggtgtgt actgcggcct 1980. gaccacaggc ttcgtgccca tgtatgtggg tgaagtgtca cccacagccc ttcgtggggc cctgggcacc ctgcaccagc tgggcatcgt cgtcggcatc ctcatcgccc aggtgttcgg 2100. cctggactcc atcatgggca acaaggacct gtggcccctg ctgctgagca tcatcttcat cccggccctg ctgcagtgca tcgtgctgcc cttctgcccc gagagtcccc gcttcctgct 2220 catcaaccgc aacgaggaga accgggccaa gagtgtgcta aagaagctgc gcgggacagc tgacgtgacc catgacctgc aggagatga ggagagagt cggcagatga tgcgggaga gaaggtcacc atcctggagc tgttccgctc ccccgcctac cgccagccca tcctcatcgc 2400 tgtggtgctg cagctgtccc agcagctgtc tggcatcaac gctgtcttct attactccac 2460 2520. gagcatcttc gagaaggcgg gggtgcagca gcctgtgtat gccaccattg gctccggtat cgtcaacacg gccttcactg tcgtgtcgct gtttgtggtg gagcgagcag gccggcggac 2580. cctgcacctc ataggcctcg ctggcatggc gggttgtgcc atactcatga ccatcgcgct 2640 agcactgctg gagcagctac cctggatgtc ctatctgagc atcgtggcca tctttggctt 2700 tgtggccttc tttgaagtgg gtcctggccc catcccatgg ttcatcgtgg ctgaactctt 2760 cagccagggt ccacgtccag ctgccattgc cgttgcaggc ttctccaact ggacctcaaa 2820 tttcattgtg ggcatgtgct tccagtatgt ggagcaactg tgtggtccct acgtcttcat 2880 catcttcact gtgctcctgg ttctgttctt catcttcacc tacttcaaag ttcctgagac 2940 taaaggccgg accttcgatg agatcgcttc cggcttccgg caggggggag ccagccaaag 3000 tgacaagaca cccgaggagc tgttccatcc cctgggggct gattcccaag tgtgataatg 3060 gatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 3120 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 3180 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 3240 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 3300 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 3360 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 3420 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc 3480 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 3540 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 3600 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcattgcctg 3660 cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact 3720 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg 3780 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggcc caagttggga 3840 agaaacctgt agggcctgcg ttacccaggc tggagtgcag tggcacattt ctgctcactg 3900 caacctcctc ctccctgggt tc 3922 <210> 98 <211> 3850 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - full polynucleotide sequence of vector genome <400> 98 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 120 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtact ctggagacgc 180 gttacataaa gcttccgacc gttagtcaga gaactgtaag tgctcagagc ctggctgaca 240 atgatctgga atgaaccaga taacaacata ataaaatctc agtaaaataa tttaacagtt 300 agcttggaag ctggtcagct ctggggaaat cagggtaaat tgtgctgtca tgaactgtcc 360 cacactgaca tcggccaaag tgaatatgaa ctttggtaga tccaatgcct gttctattta 420 tttttccagt gaaaagtatt ttgatagagc ttttcatttt gtaaatacac tgagttaacc 480 aaaatatcat ggatttccgt ttgttcttaa gacatgcaac tcgtctacgg ctataccact 540 ctgaacgcgc ccgatctcgg aagacatgca actcaaatgt aaatacagta gaatattact 600 taggtagaaa ctcctggtga ttttaaaaga ttggaaaaga atatgaggaa gagttgaata 660 atgcaaattc tagtgtgtgt gctaccgaag tgaacactta atgcacagtc tacagactag 720 gacattttat cgtgtgttgt aaaattgggt agaaacttgt gtttgtgaaa actgagcatt 780 aaaaccttac agagaccgtt tcttgtttac ttttgaaaaa aaaaagagtc acgtgagcct 840 cattttgtat ttgtgtgtgt gtgtgtgtgt gtgtctcccc tcctcccagc gtgtgtgtgc 900 tgggaggagg ggagacccca gaacaatgtc ctgcctccaa accttctcaa taggcggaag 960 ccactggctt cctccctttc ctgtctcccg tgctccagca atgcagatgg aagggaccga 1020 agggatggga gagagagccc aaccatcccc agatctgtcc ttgtcacaac ctgcctccca 1080 cctctaatgc ccccccttcc agagacttcc aggccacacc catcccgggc ttgtgggggc 1140 tggacacggg aggactacag gcgacaactc ttcccaccct ctctccctgc cacccctcct 1200 accctaacca tcatttcctc ttcctcccca gcaccgaggt gcactgagct ggacaggctg 1260 aacactcaga cccacagcaa ctgaccccgg gcccagctgg ccttggctgg cccagggcag 1320 cttccagagt gccaccatgg agcccagcag caagaagctg acgggtcgcc tcatgctggc 1380 cgtgggagga gcagtgcttg gctccctgca gtttggctac aacactggag tcatcaatgc cccccagaag gtgatcgagg agttctacaa ccagacatgg gtccaccgct atggggagag catcctgccc accacgctca ccacgctctg gtccctctca gtggccatct tttctgttgg 1560 gggcatgatt ggctccttct ctgtgggcct tttcgttaac cgctttggcc ggcggaattc 1620 aatgctgatg atgaacctgc tggccttcgt gtccgccgtg ctcatgggct tctcgaaact 1680. gggcaagtcc tttgagatgc tgatcctggg ccgcttcatc atcggtgtgt actgcggcct 1740 gaccacaggc ttcgtgccca tgtatgtggg tgaagtgtca cccacagccc ttcgtggggc cctgggcacc ctgcaccagc tgggcatcgt cgtcggcatc ctcatcgccc aggtgttcgg 1860. cctggactcc atcatgggca acaaggacct gtggcccctg ctgctgagca tcatcttcat cccggccctg ctgcagtgca tcgtgctgcc cttctgcccc gagagtcccc gcttcctgct catcaaccgc aacgaggaga accgggccaa gagtgtgcta aagaagctgc gcgggacagc tgacgtgacc catgacctgc aggagatga ggagagagt cggcagatga tgcgggaga gaaggtcacc atcctggagc tgttccgctc ccccgcctac cgccagccca tcctcatcgc 2160 tgtggtgctg cagctgtccc agcagctgtc tggcatcaac gctgtcttct attactccac 2220 gagcatcttc gagaaggcgg gggtgcagca gcctgtgtat gccaccattg gctccggtat 2280 cgtcaacacg gccttcactg tcgtgtcgct gtttgtggtg gagcgagcag gccggcggac 2340 cctgcacctc ataggcctcg ctggcatggc gggttgtgcc atactcatga ccatcgcgct 2400 agcactgctg gagcagctac cctggatgtc ctatctgagc atcgtggcca tctttggctt 2460 tgtggccttc tttgaagtgg gtcctggccc catcccatgg ttcatcgtgg ctgaactctt 2520 cagccagggt ccacgtccag ctgccattgc cgttgcaggc ttctccaact ggacctcaaa 2580 tttcattgtg ggcatgtgct tccagtatgt ggagcaactg tgtggtccct acgtcttcat 2640 catcttcact gtgctcctgg ttctgttctt catcttcacc tacttcaaag ttcctgagac 2700 taaaggccgg accttcgatg agatcgcttc cggcttccgg caggggggag ccagccaaag 2760 tgacaagaca cccgaggagc tgttccatcc cctgggggct gattcccaag tgtgataatg 2820 gatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 2880 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 2940 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 3000 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 3060 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 3120 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 3180 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc 3240 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 3300 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 3360 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcattgcctg 3420 cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact 3480 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg 3540 tccttctata atattatggg gtggagggg gtggtatgga gcaaggggcc caagttggga 3600 agaaacctgt agggcctgcg ttacccaggc tggagtgcag tggcacattt ctgctcactg 3660 caacctcctc ctccctgggt tctacgtaga taagtagcat ggcgggttaa tcattaacta 3720 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga 3780 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga 3840 gcgagcgcgc 3850 <210> 99 <211> 3514 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - part of expression cassette <400> 99 ctctggagac gcgttacata aagcttccga ccgttagtca gagaactgta agtgctcaga 60 gcctggctga caatgatctg gaatgaacca gataacaaca tataaaatc tcagtaaaat 120 aatttaacag ttagcttgga agctggtcag ctctggggaa atcagggtaa attgtgctgt 180 catgaactgt cccacactga catcggccaa agtgaatatg aactttggta gatccaatgc 240 ctgttctatt tatttttcca gtgaaaagta ttttgataga gcttttcatt ttgtaaatac 300 actgagttaa ccaaaatatc atggatttcc gtttgttctt aagacatgca actcgtctac 360 ggctatacca ctctgaacgc gcccgatctc ggaagacatg caactcaaat gtaaatacag 420 tagaatatta cttaggtaga aactcctggt gattttaaaa gattggaaaa gaatatgagg 480 aagagttgaa taatgcaaat tctagtgtgt gtgctaccga agtgaacact taatgcacag 540 tctacagact aggacatttt atcgtgtgtt gtaaaattgg gtagaaactt gtgtttgtga 600 aaactgagca ttaaaacctt acagagaccg tttcttgttt acttttgaaa aaaaaaagag 660 tcacgtgagc ctcattttgt atttgtgtgt gtgtgtgtgt gtgtgtctcc cctctccca 720 gcgtgtgtgt gctgggagga ggggagaccc cagaacaatg tcctgcctcc aaacctttc 780 aataggcgga agccactggc ttcctccctt tcctgtctcc cgtgctccag caatgcagat 840 ggaagggacc gaaggatgg gagagagagc ccaacatcc ccagatctgt ccttgtcaca 900 acctgcctcc cacctctaat gccccccctt ccagagactt ccaggccaca cccatcccgg 960 gcttgtgggg gctggacacg ggaggactac aggcgacaac tcttcccacc ctctctccct 1020 gccacccctc ctaccctaac catcatttcc tcttcctccc cagcaccgag gtgcactgag 1080 ctggacaggc tgaacactca gacccacagc aactgacccc gggcccagct ggccttggct 1140 ggcccagggc agcttccaga gtgccaccat ggagcccagc agcaagaagc tgacgggtcg 1200 cctcatgctg gccgtgggag gagcagtgct tggctccctg cagtttggct acaacactgg 1260 agtcatcaat gccccccaga aggtgatcga ggagttctac aaccagacat gggtccaccg 1320 ctatggggag agcatcctgc ccaccacgct caccacgctc tggtccctct cagtggccat 1380 cttttctgtt gggggcatga ttggctcctt ctctgtgggc cttttcgtta accgctttgg 1440 ccggcggaat tcaatgctga tgatgaacct gctggccttc gtgtccgccg tgctcatggg 1500 cttctcgaaa ctgggcaagt cctttgagat gctgatcctg ggccgcttca tcatcggtgt 1560 gtactgcggc ctgaccacag gcttcgtgcc catgtatgtg ggtgaagtgt cacccacagc 1620 ccttcgtggg gccctgggca ccctgcacca gctgggcatc gtcgtcggca tcctcatcgc 1680 ccaggtgttc ggcctggact ccatcatggg caacaaggac ctgtggcccc tgctgctgag 1740 catcatcttc atcccggccc tgctgcagtg catcgtgctg cccttctgcc ccgagagtcc 1800 ccgcttcctg ctcatcaacc gcaacgagga gaaccgggcc aagagtgtgc taaagaagct 1860 gcgcgggaca gctgacgtga cccatgacct gcaggagatg aaggaagaga gtcggcagat 1920 gatgcgggag aagaaggtca ccatcctgga gctgttccgc tccccgcct accgccagcc 1980 catcctcatc gctgtggtgc tgcagctgtc ccagcagctg tctggcatca acgctgtctt 2040 ctattactcc acgagcatct tcgagaaggc gggggtgcag cagcctgtgt atgccaccat 2100 tggctccggt atcgtcaaca cggccttcac tgtcgtgtcg ctgtttgtgg tggagcgagc 2160 aggccggcgg accctgcacc tcataggcct cgctggcatg gcgggttgtg catactcat 2220 gaccatcgcg ctagcactgc tggagcagct accctggatg tcctatctga gcatcgtggc 2280 catctttggc tttgtggcct tctttgaagt gggtcctggc cccatcccat ggttcatcgt 2340 ggctgaactc ttcagccagg gtccacgtcc agctgccatt gccgttgcag gcttctccaa 2400 ctggacctca aatttcattg tgggcatgtg cttccagtat gtggagcaac tgtgtggtcc 2460 ctacgtcttc atcatcttca ctgtgctcct ggttctgttc ttcatcttca cctacttcaa 2520 agttcctgag actaaaggcc ggaccttcga tgagatcgct tccggcttcc ggcagggggg 2580 agccagccaa agtgacaaga cacccgagga gctgttccat cccctggggg ctgattccca 2640 agtgtgataa tggatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt 2700 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct 2760 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt 2820 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac 2880 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct 2940 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca 3000 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt 3060 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc 3120 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct 3180 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg 3240 catcattgcc tgcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg 3300 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg 3360 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg 3420 cccaagttgg gaagaaacct gtagggcctg cgttacccag gctggagtgc agtggcacat 3480 ttctgctcac tgcaacctcc tcctccctgg gttc 3514 <210> 100 <211> 3010 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - full polynucleotide sequence of vector genome <400> 100 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg 60 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag 120 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtact ctggagacgc 180 gttacataac cattttgcta gagaaggccg cggaggctca gagaggtgcg caacacttgcc 240 ctgagtcaca cagcgaatgc cctccgcggt cccaacgcag agagaacgag ccgatcggca 300 gcctgagcga ggcagtggtt aggggggcc ccggccccgg ccactcccct caccccctcc 360 ccgcagagcg ccgcccagga caggctgggc cccaggcccc gccccgaggt cctgcccaca 420 cacccctgac acaccggcgt cgccagccaa tggccggggt cctataaacg ctacggtccg 480 cgcgctctct gccaccatgg agcccagcag caagaagctg acgggtcgcc tcatgctggc 540 cgtgggagga gcagtgcttg gctccctgca gtttggctac aacactggag tcatcaatgc 600 cccccagaag gtgatcgagg agttctacaa ccagacatgg gtccaccgct atggggagag 660 catcctgccc accacgctca ccacgctctg gtccctctca gtggccatct tttctgttgg 720 gggcatgatt ggctccttct ctgtgggcct tttcgttaac cgctttggcc ggcggaattc 780 aatgctgatg atgaacctgc tggccttcgt gtccgccgtg ctcatgggct tctcgaaact 840 gggcaagtcc tttgagatgc tgatcctggg ccgcttcatc atcggtgtgt actgcggcct 900 gaccacaggc ttcgtgccca tgtatgtggg tgaagtgtca cccacagccc ttcgtggggc 960 cctgggcacc ctgcaccagc tgggcatcgt cgtcggcatc ctcatcgccc aggtgttcgg 1020 cctggactcc atcatgggca acaaggacct gtggcccctg ctgctgagca tcatcttcat 1080 cccggccctg ctgcagtgca tcgtgctgcc cttctgcccc gagagtcccc gcttcctgct 1140 catcaaccgc aacgaggaga accgggccaa gagtgtgcta aagaagctgc gcgggacagc tgacgtgacc catgacctgc aggagatga ggagagagt cggcagatga tgcgggaga gaaggtcacc atcctggagc tgttccgctc ccccgcctac cgccagccca tcctcatcgc 1320 tgtggtgctg cagctgtccc agcagctgtc tggcatcaac gctgtcttct attactccac 1380. gagcatcttc gagaaggcgg gggtgcagca gcctgtgtat gccaccattg gctccggtat 1440 cgtcaacacg gccttcactg tcgtgtcgct gtttgtggtg gagcgagcag gccggcggac cctgcacctc ataggcctcg ctggcatggc gggttgtgcc atactcatga ccatcgcgct agcactgctg gagcagctac cctggatgtc ctatctgagc atcgtggcca tctttggctt 1620 tgtggccttc tttgaagtgg gtcctggccc catcccatgg ttcatcgtgg ctgaactctt 1680 cagccagggt ccacgtccag ctgccattgc cgttgcaggc ttctccaact ggacctcaaa 1740 tttcattgtg ggcatgtgct tccagtatgt ggagcaactg tgtggtccct acgtcttcat 1800 catcttcact gtgctcctgg ttctgttctt catcttcacc tacttcaaag ttcctgagac 1860 taaaggccgg accttcgatg agatcgcttc cggcttccgg caggggggag ccagccaaag 1920 tgacaagaca cccgaggagc tgttccatcc cctgggggct gattcccaag tgtgataatg 1980 gatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct 2040 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt 2100 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg 2160 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact 2220 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct 2280 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg 2340 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc 2400 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc 2460 aatccagcgg accttcttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt 2520 cgcttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcattgcctg 2580 cccgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact 2640 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg 2700 2760 agaaacctgt agggcctgcg ttacccaggc tggagtgcag tggcacattt ctgctcactg 2820 caacctcctc ctccctgggt tctacgtaga tagtagcat ggcgggttaa tcattaacta 2880 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga 2940 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga 3000 gcgagcgcgc 3010 <210> 101 <211> 2611 <212> DNA <213> Artificial Sequence <220> <223> Made in Lab - part of expression cassette <400> 101 ctctggagac gcgttacata accattttgc tagagaaggc cgcggaggct cagagaggtg 60 cgcacacttg ccctgagtca cacagcgaat gccctccgcg gtcccaacgc agagagaacg 120 agccgatcgg cagcctgagc gaggcagtgg ttaggggggg ccccggcccc ggccactccc 180 ctcaccccct ccccgcagag cgccgcccag gacaggctgg gccccaggcc ccgccccgag 240 gtcctgccca cacacccctg acacaccggc gtcgccagcc aatggccggg gtcctataaa 300 cgctacggtc cgcgcgctct ctgccaccat ggagcccagc agcaagaagc tgacgggtcg 360 cctcatgctg gccgtgggag gagcagtgct tggctccctg cagtttggct acaacactgg 420 agtcatcaat gccccccaga aggtgatcga ggagttctac aaccagacat gggtccaccg 480 ctatggggag agcatcctgc ccaccacgct caccacgctc tggtccctct cagtggccat 540 cttttctgtt gggggcatga ttggctcctt ctctgtgggc cttttcgtta accgctttgg 600 ccggcggaat tcaatgctga tgatgaacct gctggccttc gtgtccgccg tgctcatggg 660 cttctcgaaa ctgggcaagt cctttgagat gctgatcctg ggccgcttca tcatcggtgt 720 gtactgcggc ctgaccacag gcttcgtgcc catgtatgtg ggtgaagtgt cacccacagc 780 ccttcgtggg gccctgggca ccctgcacca gctgggcatc gtcgcggca tcctcatcgc 840 ccaggtgttc ggcctggact ccatcatggg caacaaggac ctgtggcccc tgctgctgag 900 catcatcttc atcccggccc tgctgcagtg catcgtgctg cccttctgcc ccgagagtcc 960 ccgcttcctg ctcatcaacc gcaacgagga gaaccgggcc aagagtgtgc taaagaagct 1020 gcgcgggaca gctgacgtga cccatgacct gcaggagatg aaggaagaga gtcggcagat 1080 gatgcgggag aagaaggtca ccatcctgga gctgttccgc tccccgcct accgccagcc 1140 catcctcatc gctgtggtgc tgcagctgtc ccagcagctg tctggcatca acgctgtctt 1200 ctattactcc acgagcatct tcgagaaggc gggggtgcag cagcctgtgt atgccaccat 1260 tggctccggt atcgtcaaca cggccttcac tgtcgtgtcg ctgtttgtgg tggagcgagc 1320 aggccggcgg accctgcacc tcataggcct cgctggcatg gcgggttgtg ccatactcat 1380 gaccatcgcg ctagcactgc tggagcagct accctggatg tcctatctga gcatcgtggc 1440 catctttggc tttgtggcct tctttgaagt gggtcctggc cccatcccat ggttcatcgt 1500 ggctgaactc ttcagccagg gtccacgtcc agctgccatt gccgttgcag gcttctccaa 1560 ctggacctca aatttcattg tgggcatgtg cttccagtat gtggagcaac tgtgtggtcc 1620 ctacgtcttc atcatcttca ctgtgctcct ggttctgttc ttcatcttca cctacttcaa 1680 agttcctgag actaaaggcc ggaccttcga tgagatcgct tccggcttcc ggcagggggg 1740 agccagccaa agtgacaaga cacccgagga gctgttccat cccctggggg ctgattccca 1800 agtgtgataa tggatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt 1860 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct 1920 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt 1980 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gttgctgac 2040 gcaaccccca ctggttgggg cattgccacc acctgtcagc tctttccgg gactttcgct 2100 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca 2160 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt 2220 ccttggctgc tcgcctgtgt tgccacctg attctgcgcg ggacgtcctt ctgctacgtc 2280 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct 2340 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctcccg 2400 catcattgcc tgcccgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg 2460 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcatttg 2520 tctgactagg tgtccttcta tatattatg gggtggaggg gggtggtatg gagcaagggg 2580 cccaagttgg gaagaaacct gtagggcctg c 2611 <210> 102 <211> 302 <212> DNA <213> Homo sapiens <400> 102 accattttgc tagagaaggc cgcggaggct cagagaggtg cgcacacttg ccctgagtca 60 cacagcgaat gccctccgcg gtcccaacgc agagagaacg agccgatcgg cagcctgagc 120 gaggcagtgg ttaggggggg ccccggcccc ggccactccc ctcaccccct ccccgcagag 180 cgccgcccag gacaggctgg gcccccaggcc ccgccccgag gtcctgccca cacaccccctg 240 acacaccggc gtcgccagcc aatggccggg gtcctataaa cgctacggtc cgcgcgctct 300 ct 302

Claims

1. An expression cassette comprising a polynucleotide sequence encoding GLUT1 or a functional variant thereof, operably ligated to a promoter, wherein the promoter is a human FLT-1 (hFLT-1) promoter sharing at least 90% identity with SEQ ID NO: 1, and the expression cassette is adjacent to 5' and 3' AAV2 inverted terminal repeats (ITRs) sharing at least 90% identity with SEQ ID NO: 6 and SEQ ID NO: 7, respectively.

2. The expression cassette according to claim 1, comprising a polyA signal, optionally containing human growth hormone (hGH) polyA.

3. An expression cassette according to claim 1 or 2, comprising a woodchuck hepatitis virus post-transcriptional regulatory element (WPRE), optionally comprising WPRE(x).

4. An expression cassette according to any one of claims 1 to 3, comprising a 3' untranslated region (3'UTR) containing a sequence that shares at least 90% identity with sequence number 4.

5. The polynucleotide sequence encoding GLUT1 is (a) is an SLC2A1 polynucleotide and / or (b) Sharing at least 90% identity with Sequence ID No. 5, An expression cassette according to any one of claims 1 to 4.

6. The expression cassette according to claim 5, wherein the SLC2A1 polynucleotide is human SLC2A1 polynucleotide.

7. An expression cassette according to any one of claims 1 to 6, which shares at least 90% identity with sequence number 12.

8. A gene therapy vector comprising an expression cassette according to any one of claims 1 to 6.

9. The vector according to claim 8, wherein the gene therapy vector is a recombinant adeno-associated virus (rAAV) vector.

10. The rAAV vector is (a) AAV6, AAV8, AAV9, AAVrh. 74, or AAVrh. 10 vector or a functional variant thereof (b) Not an AAV2 vector and / or (c) A capsid protein that shares at least 90% identity with any one of sequence numbers 77-81, The vector according to claim 9.

11. A pharmaceutical composition for treating and / or preventing a disease or disorder in a subject requiring the use of the vector according to any one of claims 8 to 10.

12. The aforementioned disease or disorder (a) It is a neurological disorder, and / or (b) Glucose transporter 1 deficiency syndrome (GLUT1DS) or Devibo disease, The pharmaceutical composition according to claim 11.

13. A pharmaceutical composition according to claim 11 or 12 for intraventricular (ICV) injection.

14. To selectively result in increased expression of the polynucleotide sequence encoding GLUT1 in the brain compared to a reference rAAV vector, and / or Selectively, this results in increased expression of GLUT1 protein in the brain, and / or increased glucose and / or lactate levels in CSF, at a level increased compared to a reference rAAV vector, wherein the increase is at least about 10% or greater. A pharmaceutical composition according to any one of claims 11 to 13.

15. The pharmaceutical composition according to any one of claims 11 to 14, wherein the dose of the vector is 1E12 vector genome / kg (vg / kg), 1E13 vg / kg, 1E14 vg / kg, or 3E14 vg / kg.

16. A pharmaceutical composition according to any one of claims 11 to 15, which causes increased glucose uptake by cerebral microvascular endothelial cells compared to a method carried out using an endogenous Glut1 promoter or a ubiquitous promoter.

17. A pharmaceutical composition for expressing GLUT1 in cells, comprising the vector according to any one of claims 8 to 10.

18. The pharmaceutical composition according to claim 17, wherein the cells are endothelial cells or neurons.

19. The aforementioned endothelial cells, (a) endothelial cells of the brain microvascular system, and / or (b) In vivo endothelial cells, The pharmaceutical composition according to claim 18.

20. The pharmaceutical composition according to claim 18, wherein the neuron is an in vivo neuron.

21. A pharmaceutical composition according to any one of claims 17 to 20 for in vivo administration to a target.

22. The pharmaceutical composition according to any one of claims 17 to 21, which causes an increase in glucose uptake by the cells compared to cells that have come into contact with a vector containing an endogenous Glut1 promoter or a ubiquitous promoter.

23. A pharmaceutical composition comprising the vector described in any one of claims 8 to 10.

24. A kit comprising a vector according to any one of claims 8 to 10 or a pharmaceutical composition according to claim 22 or 23, and optionally instructions for use.

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