LOCUS       sequence-356202-        6181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre- and Flp-dependent transgene expression (CRE-ON /
            FLP-ON) of dTom in neurons under the control of the hSyn1 promoter 
            (Kugler et al 2003).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6181)
  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 6181)
  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-dTom
FEATURES             Location/Qualifiers
     source          1..6181
                     /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        158..605
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    629..662
                     /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    708..756
                     /note="synthetic polyadenylation signal"
     polyA_signal    765..1241
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(964..983)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1276..1295)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1320..1368
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    1528..1561
                     /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(1568..1601)
                     /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(1652..1685)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1742..2446)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MAVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQT
                     AKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDG
                     GLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEI
                     HQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGR
                     HHLFLYGMDELYK"
     primer_bind     complement(1825..1848)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2023..2042)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2292..2310
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2443..2452
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2496..2529
                     /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    2580..2613
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2622..3210
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2675..2695)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3093..3104)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3218..3442
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3444..3584
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3444..3573
                     /label=AAV2 ITR
     rep_origin      3659..4114
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3746..3765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3956..3977
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4131..4150)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4250..4272
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4310..4328)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4396..4500
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4501..4569,4570..5361)
                     /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(4719..4738)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5532..6120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6021..6040
                     /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 gctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
      181 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
      241 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
      301 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
      361 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
      421 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
      481 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
      541 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
      601 cgcagtcaga tatcgtttaa actccggaga agttcctatt ctctagaaag tataggaact
      661 tccctcagca ccatggctag cggcagcctc ggagtttgaa tagatagaat aaaatatctt
      721 tattttcatt ccatctgtgt gttggttttt tgtgtgagat ctacgggtgg catccctgtg
      781 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
      841 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
      901 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
      961 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     1021 cctcctgggt tcaagcgatt ctcctgcctc aggctcccga gttgttggga ttccaggcat
     1081 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     1141 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     1201 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgcctcag ctaattgagt
     1261 aggggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     1321 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     1381 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     1441 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     1501 cagccaacgt cggggcggca ggccgtggaa gttcctattc tctagaaagt ataggaactt
     1561 caccggaata acttcgtata ggatacttta tacgaagtta tgcagaatgg tagctggatt
     1621 gtagctgcta ttagcaatat gaaacctctt aataacttcg tatagcatac attatacgaa
     1681 gttataacgc gccactagtg aattcgagct cggtacccgg ggatcctcta gagtcgactt
     1741 acttgtacag ctcgtccatg ccgtacagga acaggtggtg gcggccctcg gagcgctcgt
     1801 actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccagcttg gtgtccacgt
     1861 agtagtagcc gggcagttgc acgggcttct tggccatgta gatggtcttg aactccacca
     1921 ggtagtggcc gccgtccttc agcttcaggg cctggtggat ctcgcccttc agcacgccgt
     1981 cgcgggggta caggcgctcg gtggaggcct cccagcccat ggtcttcttc tgcattacgg
     2041 ggccgtcggg ggggaagttg gtgccgcgca tcttcacctt gtagatcagc gtgccgtcct
     2101 gcagggagga gtcctgggtc acggtcacca gaccgccgtc ctcgaagttc atcacgcgct
     2161 cccacttgaa gccctcgggg aaggacagct tcttgtaatc ggggatgtcg gcggggtgct
     2221 tcacgtacgc cttggagccg tacatgaact ggggggacag gatgtcccag gcgaagggca
     2281 gggggccgcc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct
     2341 cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga gccctccatg cgcaccttga
     2401 agcgcatgaa ctctttgatg acctcctcgc ccttgctcac ggccatggtg gcgtcgacct
     2461 gcaggcatgc aagctttact agtggtggcg ctagcataac ttcgtataaa gtatcctata
     2521 cgaagttatt tgccttaacc cagaaattat cactgttatt ctttagaatg gtgcaaagaa
     2581 taacttcgta taatgtatgc tatacgaagt tatggcgcgc caatcaacct ctggattaca
     2641 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2701 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2761 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2821 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2881 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2941 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3001 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     3061 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3121 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3181 gtcggatctc cctttgggcc gcctccccgc ctggatcctg tgccttctag ttgccagcca
     3241 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3301 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3361 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3421 ggggatgcgg tgggctctat ggcaggaacc cctagtgatg gagttggcca ctccctctct
     3481 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     3541 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta
     3601 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac
     3661 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3721 acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3781 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3841 gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca
     3901 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3961 ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa
     4021 gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4081 gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc
     4141 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     4201 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4261 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     4321 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     4381 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4441 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4501 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4561 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4621 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4681 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4741 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4801 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4861 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4921 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4981 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5041 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5101 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5161 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5221 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5281 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5341 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5401 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5461 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5521 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5581 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5641 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5701 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5761 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5821 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5881 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5941 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6001 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6061 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6121 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6181 t
//
