LOCUS       sequence-337194-        4578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone for Cre- and Flp-dependent transgene expression 
            (CRE-OFF / FLP-ON) 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 4578)
  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 4578)
  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-VTKS6
FEATURES             Location/Qualifiers
     source          1..4578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        157..604
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(628..661)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    662..695
                     /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    791..847
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    complement(858..891)
                     /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)"
     protein_bind    complement(976..1009)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1018..1606
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1071..1091)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1489..1500)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1614..1838
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1840..1980
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1840..1969
                     /label=AAV2 ITR
     rep_origin      2055..2510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2142..2161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2352..2373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2527..2546)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2646..2668
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2706..2724)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2792..2896
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2897..2965,2966..3757)
                     /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(3115..3134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3928..4516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4417..4436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(4578,1..129)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg
       61 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      121 ggggttcctg ctagactgca gagggccctg cgtatgagtg caagtgggtt ttaggaccag
      181 gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg acaagcaccc
      241 aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg
      301 gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg
      361 cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc
      421 gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc
      481 gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg
      541 cgactcagcg ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc
      601 gcagtcagat atcgtttaaa ctccggaata acttcgtata gcatacatta tacgaagtta
      661 tgaagttcct attctctaga aagtatagga acttcgcaga atggtagctg gattgtagct
      721 gctattagca atatgaaacc tcttagaagt tcctattctt caaaaggtat aggaacttca
      781 acgcgcccta gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg
      841 caagcttggt ggctagcgaa gttcctatac tttctagaga ataggaactt cttgccttaa
      901 cccagaaatt atcactgtta ttctttagaa tggtgcaaag agaagttcct ataccttttg
      961 aagaatagga acttcataac ttcgtatagc atacattata cgaagttatg gcgcgccaat
     1021 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     1081 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     1141 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     1201 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     1261 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     1321 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     1381 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     1441 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     1501 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     1561 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg atcctgtgcc
     1621 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1681 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1741 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1801 caatagcagg catgctgggg atgcggtggg ctctatggca ggaaccccta gtgatggagt
     1861 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     1921 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     1981 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     2041 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2101 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     2161 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2221 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     2281 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2341 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     2401 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     2461 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     2521 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     2581 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     2641 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     2701 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     2761 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2821 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2881 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2941 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3001 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3061 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3121 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3181 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3241 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3301 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3361 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3421 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3481 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3541 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3601 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3661 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3721 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3781 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3841 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3901 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3961 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4021 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4081 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4141 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4201 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4261 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4321 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4381 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4441 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4501 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4561 gccttttgct cacatgtc
//
