LOCUS       sequence-355057-        6225 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for Cre- and Flp-dependent transgene expression (CRE-OFF 
            / FLP-ON) of ChR2-YFP 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 6225)
  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 6225)
  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-ChR2-YFP
FEATURES             Location/Qualifiers
     source          1..6225
                     /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    complement(629..662)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    663..696
                     /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)"
     CDS             complement(join(798..1511,1512..1514,1515..1517))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(843..864)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1192..1211
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1451..1472
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1514..1523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1533..2459)
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      2456..2465
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2506..2539)
                     /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(2624..2657)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2666..3254
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2719..2739)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3137..3148)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3262..3486
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3488..3628
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3488..3617
                     /label=AAV2 ITR
     rep_origin      3703..4158
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3790..3809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4000..4021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4175..4194)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4294..4316
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4354..4372)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4440..4544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4545..4613,4614..5405)
                     /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(4763..4782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5576..6164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6065..6084
                     /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 actccggaat aacttcgtat agcatacatt atacgaagtt
      661 atgaagttcc tattctctag aaagtatagg aacttcgcag aatggtagct ggattgtagc
      721 tgctattagc aatatgaaac ctcttagaag ttcctattct tcaaaaggta taggaacttc
      781 aacgcgccct agaattctta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
      841 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagggcg
      901 gactggtagc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
      961 ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg
     1021 aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg
     1081 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     1141 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     1201 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac
     1261 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg cgaagcactg caggccgtag
     1321 ccgaaggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac
     1381 ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg
     1441 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     1501 tcgcccttgc tcaccatggt ggcggccgct ggcacggctc cggcctcggc ttcgtcttcg
     1561 acgagagtct cgacctcgat ctccgttccg ccgatgttca gtttggtggt tttgcggata
     1621 tctccgtgaa tcaatatgtg ctcgtggatc aggacgcgca ggtagtgtcc caacaacccc
     1681 caacaatttt tactcatcag atcaataatc gtgtgaccta cggtggagcc atagacgctc
     1741 aggacgccaa aaccttcggg ccccaaaatg aagagaattg ggaacatacc ccagctcacg
     1801 aaaaacagcc atgccatgcc ggtcacgacc tggcggcacc gaccctttgg cacagtatga
     1861 taaccctcga tatatgcttt ggcggcgtga aaaaatgtgt tcgcgccata gcacaatcca
     1921 agacaaaaga agatgacttt aacatagccg gttgccatgg cgctggtagc cccccacacg
     1981 atagtcccga tgtctgagac aaggagtccc atggttctcc tgctgtagtc gttgctcagg
     2041 ccggtgaggt tgctcaggcg gataaggatg acaggacaag tgagcagcca ctctgcatag
     2101 cgcagccact gcacccggtg tcctgtggca aggtagagca tagagggatt cttaaactca
     2161 aaaaagaact cgagaatcac cttaaccatt tcaatggcgc acacatagat ctcctcccag
     2221 ccgcatgtag atttccaggt ttggtaggca tagaacatca gcagcaaaat gctgaatcct
     2281 gctgcaagcc actgcaggac atttgacgcg gtctgagcgc cgttcgtgcc gcgagattca
     2341 atccatccgg cacagtaaca ttgatcctca gggaccagga cggacccgtt caccaccaca
     2401 ggattagtaa cgaacaaaag ttcgcgtccg acggcagaca aagcgccgcc atagtccatg
     2461 gtggctccgg agtcgacctg caggcatgca agcttggtgg ctagcgaagt tcctatactt
     2521 tctagagaat aggaacttct tgccttaacc cagaaattat cactgttatt ctttagaatg
     2581 gtgcaaagag aagttcctat accttttgaa gaataggaac ttcataactt cgtatagcat
     2641 acattatacg aagttatggc gcgccaatca acctctggat tacaaaattt gtgaaagatt
     2701 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2761 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2821 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     2881 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     2941 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3001 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3061 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3121 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3181 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3241 ggccgcctcc ccgcctggat cctgtgcctt ctagttgcca gccatctgtt gtttgcccct
     3301 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     3361 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     3421 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     3481 ctatggcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     3541 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     3601 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     3661 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     3721 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     3781 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3841 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3901 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3961 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4021 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4081 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4141 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4201 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4261 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4321 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     4381 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4441 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4501 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4561 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4621 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4681 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4741 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4801 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4861 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     4921 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     4981 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5041 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5101 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5161 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5221 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5281 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5341 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5401 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5461 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5521 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5581 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5641 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5701 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5761 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5821 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5881 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5941 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6001 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6061 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6121 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6181 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
