LOCUS       sequence-272197-        6430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of ChR2-mCherry in PV cortical 
            interneuronsunder the control of the E2 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6430)
  AUTHORS   Vormstein-Schneider D, Lin JD, Pelkey KA, Chittajallu R, Guo B, 
            Arias-Garcia MA, Allaway K, Sakopoulos S, Schneider G, Stevenson O, 
            Vergara J, Sharma J, Zhang Q, Franken TP, Smith J, Ibrahim LA, M 
            Astro KJ, Sabri E, Huang S, Favuzzi E, Burbridge T, Xu Q, Guo L, 
            Vogel I, Sanchez V, Saldi GA, Gorissen BL, Yuan X, Zaghloul KA, 
            Devinsky O, Sabatini BL, Batista-Brito R, Reynolds J, Feng G, Fu Z, 
            McBain CJ, Fishell G, Dimidschstein J
  TITLE     Viral manipulation of functionally distinct interneurons in mice, 
            non-human primates and humans.
  JOURNAL   Nat Neurosci. 2020 Dec;23(12):1629-1636. doi: 
            10.1038/s41593-020-0692-9. Epub 2020 Aug 17.
  PUBMED    32807948
REFERENCE   2  (bases 1 to 6430)
  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"; doi: 
            "10.1038/s41593-020-0692-9"; journalName: "Nat Neurosci"; date: 
            "2020-12"; volume: "23"; issue: "12"; pages: "1629-1636"
COMMENT     Sequence Label: pAAV-S5E2-ChR2-mCherry
FEATURES             Location/Qualifiers
     source          1..6430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          934..1066
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1081..1090
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1087..2013
                     /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"
     CDS             2026..2733
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(2168..2186)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2436..2455
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2776..3364
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2829..2849)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3247..3258)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3533..3614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3570..3589)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3624..3643
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3693..3833
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3693..3822
                     /label=AAV2 ITR
     rep_origin      3908..4363
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3995..4014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4205..4226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4380..4399)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4499..4521
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4559..4577)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4645..4749
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4750..4818,4819..5610)
                     /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(4968..4987)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5781..6369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6270..6289
                     /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 gcggccgcac gcgtttaatt aaactagtgg tttaatctaa catggctgct
      181 atagcttact gactagaagt taagtgcaca cttcctaaaa gaaggctttg acacaagcca
      241 cttcagttcc ctcctcattt tcttgtcccc attcctctct ctgtagaatt ctgagatttc
      301 aattcagttt tatacagaaa ccacattact gtaagcccta caaagttatg gcaatatagc
      361 tatatggagt caagtaatgt aggttatttt tttcccaatg gtgctggtga aggtggcaat
      421 tatgtagcta tacttagcag actgaggaaa ttctgctaga gtcagcattt gtctcttcat
      481 tgctatgaaa cagtaatgga aaaataaaca aaaacaaaag gcaaacacta tgcataattc
      541 cctcagatca tattaacatg tgatgttgga gtaaattgtt ataaccccat tttggaaata
      601 cttaccttaa ttaactatga tttccttaaa ataatgcagt atttacaatc tatatgaaag
      661 cactatatgg gacacatggt atgatggaac agtgcaccca agagacacca agaacattcc
      721 tgtctgtggc agtcttttct ctatacagag gcatttagtc tcaattgctc agagttattt
      781 taaacttcct gcagcccggg ctgggcataa aagtcagggc agagccatct attgcttaca
      841 tttgcttcta gcctgcaagt cgaggagcgc agccttccag aagcagagcg cggcgcctta
      901 agctgcagaa gttggtcgtg aggcactggg cacgtaagta tcaaggttac aagacaggtt
      961 taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag
     1021 gcacctattg gtcttactga catccacttt gcctttctct ccacagcgag ctcgctagta
     1081 gccaccatgg actatggcgg cgctttgtct gccgtcggac gcgaactttt gttcgttact
     1141 aatcctgtgg tggtgaacgg gtccgtcctg gtccctgagg atcaatgtta ctgtgccgga
     1201 tggattgaat ctcgcggcac gaacggcgct cagaccgcgt caaatgtcct gcagtggctt
     1261 gcagcaggat tcagcatttt gctgctgatg ttctatgcct accaaacctg gaaatctaca
     1321 tgcggctggg aggagatcta tgtgtgcgcc attgaaatgg ttaaggtgat tctcgagttc
     1381 ttttttgagt ttaagaatcc ctctatgctc taccttgcca caggacaccg ggtgcagtgg
     1441 ctgcgctatg cagagtggct gctcacttgt cctgtcatcc ttatccgcct gagcaacctc
     1501 accggcctga gcaacgacta cagcaggaga accatgggac tccttgtctc agacatcggg
     1561 actatcgtgt ggggggctac cagcgccatg gcaaccggct atgttaaagt catcttcttt
     1621 tgtcttggat tgtgctatgg cgcgaacaca ttttttcacg ccgccaaagc atatatcgag
     1681 ggttatcata ctgtgccaaa gggtcggtgc cgccaggtcg tgaccggcat ggcatggctg
     1741 tttttcgtga gctggggtat gttcccaatt ctcttcattt tggggcccga aggttttggc
     1801 gtcctgagcg tctatggctc caccgtaggt cacacgatta ttgatctgat gagtaaaaat
     1861 tgttgggggt tgttgggaca ctacctgcgc gtcctgatcc acgagcacat attgattcac
     1921 ggagatatcc gcaaaaccac caaactgaac atcggcggaa cggagatcga ggtcgagact
     1981 ctcgtcgaag acgaagccga ggccggagcc gtgccagcgg ccgccgtgag caagggcgag
     2041 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     2101 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     2161 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     2221 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     2281 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     2341 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     2401 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     2461 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     2521 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     2581 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     2641 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     2701 tccaccggcg gcatggacga gctgtacaag taaaacgcgt gcccctgcag ggaattcgat
     2761 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2821 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2881 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2941 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3001 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3061 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3121 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3181 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3241 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3301 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3361 ccgcatcgat accgagcgct gctcgagaga tctgtgatag cggccatcaa gctggccgcg
     3421 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3481 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3541 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3601 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cagcttatcg
     3661 ataccgcatg cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     3721 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     3781 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     3841 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     3901 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3961 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     4021 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4081 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4141 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4201 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     4261 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     4321 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     4381 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     4441 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     4501 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     4561 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4621 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4681 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4741 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4801 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4861 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4921 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4981 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5041 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5101 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5161 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5221 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5281 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5341 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5401 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5461 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5521 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5581 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5641 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5701 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5761 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5821 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5881 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     5941 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6001 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6061 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6121 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6181 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6241 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6301 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6361 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6421 gctcacatgt
//
