LOCUS       sequence-337193-        4596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone for Cre- and Flp-dependent transgene expression (CRE-ON
            / FLP-OFF) 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 4596)
  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 4596)
  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-VTKS5
FEATURES             Location/Qualifiers
     source          1..4596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   36..165
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        193..640
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    664..697
                     /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(704..737)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(788..821)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    836..892
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    912..945
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    996..1029
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1030..1063
                     /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    1072..1660
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1125..1145)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1543..1554)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1668..1892
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1894..2034
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1894..2023
                     /label=AAV2 ITR
     rep_origin      2109..2564
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2196..2215)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2406..2427
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2581..2600)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2700..2722
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2760..2778)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2846..2950
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2951..3019,3020..3811)
                     /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(3169..3188)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3982..4570
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4471..4490
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcctggcc ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc
       61 gctcactgag gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc
      121 gagcgcgcag agagggagtg gccaactcca tcactagggg ttcctgctag actgcagagg
      181 gccctgcgta tgagtgcaag tgggttttag gaccaggatg aggcggggtg ggggtgccta
      241 cctgacgacc gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca
      301 tcccctatca gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc
      361 ttcagcaccg cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag
      421 cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct
      481 tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg
      541 ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc
      601 ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag tcagatatcg tttaaactcc
      661 ggagaagttc ctattctcta gaaagtatag gaacttctcc ggaataactt cgtataggat
      721 actttatacg aagttatgca gaatggtagc tggattgtag ctgctattag caatatgaaa
      781 cctcttaata acttcgtata gcatacatta tacgaagtta taacgcgcca ctagtgaatt
      841 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc tttactagtg
      901 gtggcgctag cataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga
      961 aattatcact gttattcttt agaatggtgc aaagaataac ttcgtataat gtatgctata
     1021 cgaagttatg aagttcctat tctctagaaa gtataggaac ttcggcgcgc caatcaacct
     1081 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     1141 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     1201 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     1261 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     1321 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     1381 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     1441 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
     1501 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     1561 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     1621 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggatcctg tgccttctag
     1681 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1741 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1801 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1861 caggcatgct ggggatgcgg tgggctctat ggcaggaacc cctagtgatg gagttggcca
     1921 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     1981 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     2041 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     2101 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2161 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     2221 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2281 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     2341 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2401 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     2461 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     2521 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     2581 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     2641 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2701 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     2761 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2821 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2881 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2941 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3001 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3061 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3121 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3181 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3241 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3301 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3361 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3421 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3481 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3541 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3601 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3661 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3721 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3781 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3841 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3901 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3961 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4021 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4081 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4141 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4201 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4261 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4321 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4381 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4441 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4501 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4561 cctatggaaa aacgccagca acgcggcctt tttacg
//
