LOCUS Exported 5286 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION An AAV genome that expresses tTA from the hSyn1 promoter. ACCESSION . VERSION . KEYWORDS pAAV-hSyn1-tTA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5286) AUTHORS Chan KY, Jang MJ, Yoo BB, Greenbaum A, Ravi N, Wu WL, Sanchez-Guardado L, Lois C, Mazmanian SK, Deverman BE, Gradinaru V TITLE Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. JOURNAL Nat Neurosci. 2017 Aug;20(8):1172-1179. doi: 10.1038/nn.4593. Epub 2017 Jun 26. PUBMED 28671695 REFERENCE 2 (bases 1 to 5286) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5286 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 175..622 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 1269..1379 /codon_start=1 /product="3 tandem repeats of the minimal activation domain of herpes simplex virus VP16 (Cheng et al., 2013)" /label=VP48 /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML" misc_feature 1413..2001 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1466..1486) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1884..1895) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2033..2509 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2232..2251) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 2549..2689 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2549..2678 /label=AAV2 ITR rep_origin 2764..3219 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2851..2870) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3061..3082 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3236..3255) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3355..3377 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3415..3433) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3501..3605 /gene="bla" /label=AmpR promoter CDS 3606..4466 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3824..3843) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4637..5225 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5126..5145 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca 181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga 241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg 301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt 361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac 421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc 481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca 541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt 601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccgccac catgagtcgg ctggataaat 661 ctaaagtcat aaacggcgct ctggaattac tcaatgaagt cggtatcgaa ggcctgacga 721 caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg cacgtgaaga 781 acaagcgggc cctgctcgat gccctggcca tcgagatgct ggacaggcat catacccact 841 tctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc aagtcattcc 901 gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag 961 agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc 1021 tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc tgcgtattgg 1081 aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc gattctatgc 1141 ccccacttct gagacaagca attgagctgt tcgaccggca gggagccgaa cctgccttcc 1201 ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg 1261 ggccggccga cgcccttgac gattttgact tagacatgct cccagccgat gcccttgacg 1321 actttgacct tgatatgctg cctgctgacg ctcttgacga ttttgacctt gacatgctcc 1381 ccgggtaaga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg 1441 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 1501 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata 1561 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg 1621 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc 1681 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct 1741 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt 1801 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg 1861 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc 1921 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct 1981 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca 2041 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac 2101 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa 2161 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag 2221 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc 2281 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt 2341 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac 2401 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc 2461 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta 2521 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc 2581 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc 2641 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg 2701 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat 2761 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 2821 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg 2881 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 2941 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg 3001 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 3061 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt 3121 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat 3181 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa 3241 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc 3301 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga 3361 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg 3421 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg 3481 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 3541 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 3601 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc 3661 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg 3721 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc 3781 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat 3841 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg 3901 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag 3961 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa 4021 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc 4081 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca 4141 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc 4201 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc 4261 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg 4321 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta 4381 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag 4441 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga 4501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 4561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 4621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 4681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 4741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt 4801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 4861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 4921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 4981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 5041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 5101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 5161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 5221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 5281 acatgt //