LOCUS       sequence-355103-        6633 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 cortical interneurons under the control of 
            the Dlx enhancer (Dimidschstein et al 2016).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6633)
  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 6633)
  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-VTKD6-ChR2-YFP
FEATURES             Location/Qualifiers
     source          1..6633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          862..994
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     977..996
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    complement(1037..1070)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1071..1104
                     /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(1206..1919,1920..1922,1923..1925))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1251..1272)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1600..1619
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1859..1880
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1922..1931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1941..2867)
                     /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      2864..2873
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2914..2947)
                     /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(3032..3065)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3074..3662
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3127..3147)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3545..3556)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3670..3894
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3896..4036
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3896..4025
                     /label=AAV2 ITR
     rep_origin      4111..4566
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4198..4217)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4408..4429
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4583..4602)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4702..4724
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4762..4780)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4848..4952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4953..5021,5022..5813)
                     /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(5171..5190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5984..6572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6473..6492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgg gcgacctttg gtcgcccggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
      121 ctagactgca gagggcccgc ggccgcacgc gtttattcag aatgttatgc actcacagtg
      181 gtttggcatg catctggtga atttttttta acgaaaaatt agtgttggtt tcgatgtatg
      241 gtagcattct ccctaacgta atttgaataa ttcagcaaag ccccactacc agctgtactt
      301 ctgcagcctc ttccattctt ttcagcatta taattttggt taattttcaa ttttaggtcc
      361 tacgtctctg caatttgtgt atgaataaca gaataatttc cctcttttgt ttcgcctttc
      421 ctgttcctga atctaaataa agatggcttt ttagtattaa aagtggaaga aaattacagg
      481 taattatctt tgacggtaaa aacgctgtaa tcagcgggct acatgaaaaa ttactctaat
      541 tatggctgca tttaagagaa tggaaaaaaa ccttcttgtg gataaaaacc ttaaattgtc
      601 cccaatgtct gcttcaaatt ggatggcact gcagctggag gctttgttca gaattgatcc
      661 tggggagcta cgaacccaaa gtttcacagt aggaagctag atatcgttta aacttcctgc
      721 agcccgggct gggcataaaa gtcagggcag agccatctat tgcttacatt tgcttctagc
      781 ctgcaagtcg aggagcgcag ccttccagaa gcagagcgcg gcgccttaag ctgcagaagt
      841 tggtcgtgag gcactgggca cgtaagtatc aaggttacaa gacaggttta aggagaccaa
      901 tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt
      961 cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt caattacagc
     1021 tcttaagaaa gctagcataa cttcgtatag catacattat acgaagttat gaagttccta
     1081 ttctctagaa agtataggaa cttcgcagaa tggtagctgg attgtagctg ctattagcaa
     1141 tatgaaacct cttagaagtt cctattcttc aaaaggtata ggaacttcaa cgcgccctag
     1201 aattcttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     1261 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggtagctc
     1321 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     1381 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg
     1441 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
     1501 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     1561 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     1621 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     1681 tgctgcttca tgtggtcggg gtagcgggcg aagcactgca ggccgtagcc gaaggtggtc
     1741 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
     1801 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     1861 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     1921 accatggtgg cggccgctgg cacggctccg gcctcggctt cgtcttcgac gagagtctcg
     1981 acctcgatct ccgttccgcc gatgttcagt ttggtggttt tgcggatatc tccgtgaatc
     2041 aatatgtgct cgtggatcag gacgcgcagg tagtgtccca acaaccccca acaattttta
     2101 ctcatcagat caataatcgt gtgacctacg gtggagccat agacgctcag gacgccaaaa
     2161 ccttcgggcc ccaaaatgaa gagaattggg aacatacccc agctcacgaa aaacagccat
     2221 gccatgccgg tcacgacctg gcggcaccga ccctttggca cagtatgata accctcgata
     2281 tatgctttgg cggcgtgaaa aaatgtgttc gcgccatagc acaatccaag acaaaagaag
     2341 atgactttaa catagccggt tgccatggcg ctggtagccc cccacacgat agtcccgatg
     2401 tctgagacaa ggagtcccat ggttctcctg ctgtagtcgt tgctcaggcc ggtgaggttg
     2461 ctcaggcgga taaggatgac aggacaagtg agcagccact ctgcatagcg cagccactgc
     2521 acccggtgtc ctgtggcaag gtagagcata gagggattct taaactcaaa aaagaactcg
     2581 agaatcacct taaccatttc aatggcgcac acatagatct cctcccagcc gcatgtagat
     2641 ttccaggttt ggtaggcata gaacatcagc agcaaaatgc tgaatcctgc tgcaagccac
     2701 tgcaggacat ttgacgcggt ctgagcgccg ttcgtgccgc gagattcaat ccatccggca
     2761 cagtaacatt gatcctcagg gaccaggacg gacccgttca ccaccacagg attagtaacg
     2821 aacaaaagtt cgcgtccgac ggcagacaaa gcgccgccat agtccatggt ggctccggag
     2881 tcgacctgca ggcatgcaag cttggtggct agcgaagttc ctatactttc tagagaatag
     2941 gaacttcttg ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagagaa
     3001 gttcctatac cttttgaaga ataggaactt cataacttcg tatagcatac attatacgaa
     3061 gttatggcgc gccaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3121 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3181 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3241 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3301 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3361 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3421 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     3481 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3541 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3601 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3661 gcctggatcc tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3721 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3781 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3841 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcaggaa
     3901 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3961 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4021 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4081 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4141 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc
     4201 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4261 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4321 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4381 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4441 actctatctc gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt
     4501 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4561 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4621 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4681 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4741 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     4801 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4861 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4921 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4981 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5041 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5101 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5161 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5221 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5281 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5341 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5401 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5461 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5521 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5581 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5641 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5701 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5761 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     5821 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5881 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5941 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6001 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6061 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6121 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6181 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6241 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6301 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6361 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6421 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6481 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6541 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6601 tcctggcctt ttgctggcct tttgctcaca tgt
//
