LOCUS       sequence-355061-        6193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre- and Flp-dependent transgene expression (CRE-ON /
            FLP-ON) of GFP 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 6193)
  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 6193)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..6193
                     /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(join(1739..2452,2453..2455,2456..2458))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1742..2452,2453..2455,2456..2458))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1784..1805)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2133..2152
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2392..2413
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2455..2464
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2508..2541
                     /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    2592..2625
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2634..3222
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2687..2707)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3105..3116)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3230..3454
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3456..3596
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3456..3585
                     /label=AAV2 ITR
     rep_origin      3671..4126
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3758..3777)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3968..3989
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4143..4162)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4262..4284
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4322..4340)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4408..4512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4513..4581,4582..5373)
                     /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(4731..4750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5544..6132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6033..6052
                     /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 ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     1801 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt
     1861 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     1921 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     1981 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     2041 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     2101 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     2161 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     2221 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     2281 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     2341 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     2401 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     2461 tggcgtcgac ctgcaggcat gcaagcttta ctagtggtgg cgctagcata acttcgtata
     2521 aagtatccta tacgaagtta tttgccttaa cccagaaatt atcactgtta ttctttagaa
     2581 tggtgcaaag aataacttcg tataatgtat gctatacgaa gttatggcgc gccaatcaac
     2641 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2701 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2761 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2821 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2881 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2941 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3001 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     3061 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     3121 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     3181 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggatcc tgtgccttct
     3241 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3301 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3361 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3421 agcaggcatg ctggggatgc ggtgggctct atggcaggaa cccctagtga tggagttggc
     3481 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3541 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     3601 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     3661 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3721 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     3781 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3841 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     3901 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3961 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4021 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     4081 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     4141 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4201 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4261 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     4321 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4381 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4441 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4501 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4561 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4621 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4681 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4741 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4801 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4861 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4921 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4981 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5041 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5101 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5161 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5221 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5281 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5341 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5401 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5461 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5521 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5581 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5641 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5701 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5761 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5821 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5881 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5941 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6001 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6061 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6181 tttgctcaca tgt
//
