LOCUS       sequence-190738-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV genome that expresses tTA from the hSyn1 promoter that 
            contains a positive feedback loop for amplifed expression of the 
            tTA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5455)
  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 5455)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nn.4593"; 
            journalName: "Nat Neurosci"; date: "2017-08"; volume: "20"; issue: 
            "8"; pages: "1172-1179"
COMMENT     Sequence Label: pAAV-ihSyn1-tTA
FEATURES             Location/Qualifiers
     source          1..5455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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)"
     protein_bind    662..680
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    698..716
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1438..1548
                     /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    1582..2170
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1635..1655)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2053..2064)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2202..2678
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2401..2420)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2718..2858
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2718..2847
                     /label=AAV2 ITR
     rep_origin      2933..3388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3020..3039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3230..3251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3405..3424)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3524..3546
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3584..3602)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3670..3774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3775..3843,3844..4635)
                     /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(3993..4012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4806..5394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5295..5314
                     /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 agtcgagaac agatctctaa aacaggtaag tcccattaat
      661 ctccctatca gtgatagaga aggtctgaag agtttactcc ctatcagtga tagagattaa
      721 ttcctctact aaccttgttc atcttttctt tttttttcta caggtcctgg gtgattaaca
      781 gcttaagggc ccggatccgg taccgccacc atgagtcggc tggataaatc taaagtcata
      841 aacggcgctc tggaattact caatgaagtc ggtatcgaag gcctgacgac aaggaaactc
      901 gctcaaaagc tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc
      961 ctgctcgatg ccctggccat cgagatgctg gacaggcatc atacccactt ctgccccctg
     1021 gaaggcgagt catggcaaga ctttctgcgg aacaacgcca agtcattccg ctgtgctctc
     1081 ctctcacatc gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac
     1141 gaaaccctgg aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca
     1201 ctgtacgctc tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag
     1261 catcaagtag caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg
     1321 agacaagcaa ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg
     1381 gaactaatca tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac
     1441 gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt
     1501 gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaagaa
     1561 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     1621 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     1681 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     1741 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     1801 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     1861 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     1921 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     1981 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     2041 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     2101 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     2161 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     2221 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     2281 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     2341 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     2401 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     2461 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     2521 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     2581 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     2641 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     2701 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2761 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     2821 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     2881 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     2941 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3001 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3061 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3121 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     3181 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3241 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     3301 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3361 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     3421 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3481 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3541 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3601 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3661 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3721 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3781 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3841 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3901 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3961 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4021 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4081 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4141 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4201 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4261 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4321 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4381 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4441 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4501 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4561 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4621 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4681 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4741 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4801 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4861 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4921 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4981 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     5041 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5101 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5161 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5221 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5281 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5341 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5401 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
