LOCUS       sequence-337192-        5461 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV backbone for Cre- and Flp-dependent transgene expression (CRE-ON
            / 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 5461)
  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 5461)
  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-VTKS4
FEATURES             Location/Qualifiers
     source          1..5461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    32..256
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   258..398
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   258..387
                     /label=AAV2 ITR
     rep_origin      473..928
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(560..579)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     770..791
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(945..964)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1064..1086
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1124..1142)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1210..1314
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1315..1383,1384..2175)
                     /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(1533..1552)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2346..2934
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2835..2854
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2996..3125
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        3153..3600
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    3624..3657
                     /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)"
     polyA_signal    3703..3751
                     /note="synthetic polyadenylation signal"
     polyA_signal    3760..4236
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3959..3978)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(4271..4290)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4315..4363
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    4523..4556
                     /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(4563..4596)
                     /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(4647..4680)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4695..4751
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     protein_bind    4771..4804
                     /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    4855..4888
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(4897..5461,1..24)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4950..4970)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5368..5379)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 tctccctttg ggccgcctcc ccgcctggat cctgtgcctt ctagttgcca gccatctgtt
       61 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
      121 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
      181 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
      241 gcggtgggct ctatggcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
      301 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
      361 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
      421 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
      481 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
      541 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
      601 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
      661 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
      721 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
      781 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
      841 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
      901 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
      961 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     1021 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     1081 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     1141 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     1201 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1261 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     1321 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     1381 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1441 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1501 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1561 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1621 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1681 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1741 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1801 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1861 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1921 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1981 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2041 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2101 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2161 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     2221 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     2281 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     2341 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     2401 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     2461 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     2521 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     2581 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2641 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2701 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2761 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2821 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2881 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2941 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtcctgc
     3001 aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga cctttggtcg
     3061 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
     3121 ttcctgctag actgcagagg gccctgcgta tgagtgcaag tgggttttag gaccaggatg
     3181 aggcggggtg ggggtgccta cctgacgacc gaccccgacc cactggacaa gcacccaacc
     3241 cccattcccc aaattgcgca tcccctatca gagaggggga ggggaaacag gatgcggcga
     3301 ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc cttcgccccc gcctggcggc
     3361 gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa
     3421 actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac
     3481 cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac
     3541 tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag
     3601 tcagatatcg tttaaactcc ggagaagttc ctattctcta gaaagtatag gaacttccct
     3661 cagcaccatg gctagcggca gcctcggagt ttgaatagat agaataaaat atctttattt
     3721 tcattccatc tgtgtgttgg ttttttgtgt gagatctacg ggtggcatcc ctgtgacccc
     3781 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     3841 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     3901 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     3961 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     4021 tgggttcaag cgattctcct gcctcaggct cccgagttgt tgggattcca ggcatgcatg
     4081 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     4141 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     4201 ttacaggcgt gaaccactgc tcccttccct gtccttctgc ctcagctaat tgagtagggg
     4261 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     4321 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4381 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4441 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4501 aacgtcgggg cggcaggccg tggaagttcc tattctctag aaagtatagg aacttcaccg
     4561 gaataacttc gtataggata ctttatacga agttatgcag aatggtagct ggattgtagc
     4621 tgctattagc aatatgaaac ctcttaataa cttcgtatag catacattat acgaagttat
     4681 aacgcgccac tagtgaattc gagctcggta cccggggatc ctctagagtc gacctgcagg
     4741 catgcaagct ttactagtgg tggcgctagc ataacttcgt ataaagtatc ctatacgaag
     4801 ttatttgcct taacccagaa attatcactg ttattcttta gaatggtgca aagaataact
     4861 tcgtataatg tatgctatac gaagttatgg cgcgccaatc aacctctgga ttacaaaatt
     4921 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4981 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     5041 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     5101 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     5161 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     5221 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     5281 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     5341 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     5401 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     5461 a
//
