LOCUS       sequence-337191-        4510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone for Flp-dependent transgene expression of any transgene
            in neurons under the control of the hSyn1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4510)
  AUTHORS   Pouchelon G, Vergara J, McMahon J, Gorissen BL, Lin JD, 
            Vormstein-Schneider D, Niehaus JL, Burbridge TJ, Wester JC, Sherer 
            M, Fernandez-Otero M, Allaway KC, Pelkey K, Chittajallu R, McBain 
            CJ, Fan M, Nasse JS, Wildenberg GA, Fishell G, Dimidschstein J
  TITLE     A versatile viral toolkit for functional discovery in the nervous 
            system.
  JOURNAL   Cell Rep Methods. 2022 May 26;2(6):100225. doi: 
            10.1016/j.crmeth.2022.100225. eCollection 2022 Jun 20.
  PUBMED    35784651
REFERENCE   2  (bases 1 to 4510)
  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"; journalName: "Cell Rep 
            Methods."; date: "2022-05-26"; volume: "2(6):100225. doi"; pages: " 
            10.1016/j.crmeth.2022.100225. eCollection 2022 Jun 20"
COMMENT     Sequence Label: pAAV-VTKS3
FEATURES             Location/Qualifiers
     source          1..4510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        103..550
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    574..607
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    703..759
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    complement(770..803)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    896..1484
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(949..969)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1367..1378)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1492..1716
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1718..1858
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1718..1847
                     /label=AAV2 ITR
     rep_origin      1933..2388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2020..2039)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2230..2251
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2405..2424)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2524..2546
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2584..2602)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2670..2774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2775..2843,2844..3635)
                     /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(2993..3012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3806..4394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4295..4314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(4456..4510,1..75)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
       61 tcactagggg ttcctgctag actgcagagg gccctgcgta tgagtgcaag tgggttttag
      121 gaccaggatg aggcggggtg ggggtgccta cctgacgacc gaccccgacc cactggacaa
      181 gcacccaacc cccattcccc aaattgcgca tcccctatca gagaggggga ggggaaacag
      241 gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc cttcgccccc
      301 gcctggcggc gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg
      361 tcccccgcaa actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc
      421 cggaccgcac cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg
      481 cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct
      541 gagagcgcag tcagatatcg tttaaactcc ggagaagttc ctattctcta gaaagtatag
      601 gaacttcgca gaatggtagc tggattgtag ctgctattag caatatgaaa cctcttagaa
      661 gttcctattc ttcaaaaggt ataggaactt caacgcgccc tagaattcga gctcggtacc
      721 cggggatcct ctagagtcga cctgcaggca tgcaagcttg gtggctagcg aagttcctat
      781 actttctaga gaataggaac ttcttgcctt aacccagaaa ttatcactgt tattctttag
      841 aatggtgcaa agagaagttc ctataccttt tgaagaatag gaacttcggc gcgccaatca
      901 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
      961 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     1021 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     1081 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     1141 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     1201 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     1261 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     1321 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     1381 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     1441 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggat cctgtgcctt
     1501 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     1561 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     1621 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1681 atagcaggca tgctggggat gcggtgggct ctatggcagg aacccctagt gatggagttg
     1741 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     1801 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     1861 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     1921 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1981 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     2041 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2101 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     2161 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2221 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     2281 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     2341 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     2401 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     2461 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     2521 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     2581 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2641 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2701 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2761 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2821 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2881 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2941 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3001 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3061 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3121 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3181 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3241 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3301 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3361 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3421 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3481 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3541 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3601 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3661 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3721 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3781 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3841 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3901 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3961 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4021 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4081 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4141 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4201 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4261 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4321 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4381 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4441 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
     4501 cgtcgggcga
//
