LOCUS       sequence-421864-        6483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing mCherry-tagged channelrhodopsin-2 driven by
            PV+ basket cell-targeting enhancer. Alias: AAV_WDP0003_ChR2_mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6483)
  AUTHORS   Furlanis E, Dai M, Leyva Garcia B, Tran T, Vergara J, Pereira A, 
            Gorissen BL, Wills S, Vlachos A, Hairston A, Dwivedi D, Du S, 
            McMahon J, Huang S, Morabito A, Vazquez A, Kim S, Lee AT, Chang EF, 
            Razzaq T, Qazi A, Vargish G, Yuan X, Caccavano A, Hunt S, 
            Chittajallu R, McLean N, Hewitt L, Paranzino E, Rice H, Cummins AC, 
            Plotnikova A, Mohanty A, Tangen AC, Shin JH, Azadi R, Eldridge MAG, 
            Alvarez VA, Averbeck BB, Alyahyay M, Vallejo TR, Soheib M, Vattino 
            LG, MacGregor CP, Chatain CP, Banks E, Olah VJ, Naskar S, Hill S, 
            Liebergall S, Badiani R, Hyde L, Hanley E, Xu Q, Allaway KC, 
            Goldberg EM, Rowan MJM, Nowakowski TJ, Lee S, Favuzzi E, Kaeser PS, 
            Sjulson L, Batista-Brito R, Takesian AE, Ibrahim LA, Iqbal A, Pelkey
            KA, McBain CJ, Dimidschstein J, Fishell G, Wang Y
  TITLE     An enhancer-AAV toolbox to target and manipulate distinct 
            interneuron subtypes.
  JOURNAL   Neuron. 2025 May 21;113(10):1525-1547.e15. doi: 
            10.1016/j.neuron.2025.05.002.
  PUBMED    40403705
REFERENCE   2  (bases 1 to 6483)
  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: 
            "10.1016/j.neuron.2025.05"; journalName: "Neuron"; date: 
            "2025-05-21- 21"; volume: "113"; issue: "10"; pages: "1525-1547.e15"
COMMENT     Sequence Label: pAAV_BiPVe3_ChR2_mCherry
FEATURES             Location/Qualifiers
     source          1..6483
                     /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          977..1109
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1131..1140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1137..2063
                     /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             2076..2783
                     /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(2218..2236)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2486..2505
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2829..3417
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2882..2902)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3300..3311)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3586..3667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3623..3642)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3677..3696
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3746..3886
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3746..3875
                     /label=AAV2 ITR
     rep_origin      3961..4416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4048..4067)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4258..4279
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4433..4452)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4552..4574
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4612..4630)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4698..4802
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4803..4871,4872..5663)
                     /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(5021..5040)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5834..6422
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6323..6342
                     /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 gcgtatcgat agaccgctgt tgcagtattt cagggaacaa
      181 gatcagaatg gtatagccta cctgaaaaat aggcaatcct ttcaaaatta aatgcacttt
      241 ctcttgggta ggagacaaga taatctatga taagggaaga gagatgtctc cagcttttat
      301 tttttttctt gtaatttgca tttaagccct ggtaacagtt taatatgaag gtgactatgg
      361 cagctccatt tattcaatct atcaatactc tgttgtcaca ggaggagcca tgaggacttg
      421 agtgcagagg aggaatcccc acaaagcagt cttcactctt atcatagctt ccatgtgccc
      481 ttcttattca gcagctctca ttaaaggaac actatcttta gcacagggat tctacttgac
      541 attgacaagg acaggctcag tagcaaatgg gccaaaacta tgaataggcg attcacagca
      601 gagtaaaatc caatggcagc agacataaaa agaagctcaa aatgacaaac aagtcagaca
      661 aacacaaatt caagtaacaa taaaaaatgg agcactggtt tgactcgtac acaagcaaac
      721 agacacaaac aaaacaaaag agtcattaca cctggggcta aagacttctc ttatgtattg
      781 caaagtggaa ataagacctc caggcaaaat gtgaatcgat gtttaaactt cctgcagccc
      841 gggctgggca taaaagtcag ggcagagcca tctattgctt acatttgctt ctagcctgca
      901 agtcgaggag cgcagccttc cagaagcaga gcgcggcgcc ttaagctgca gaagttggtc
      961 gtgaggcact gggcacgtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
     1021 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
     1081 tgacatccac tttgcctttc tctccacagc gagctcgcta gcgcgatcgc gccaccatgg
     1141 actatggcgg cgctttgtct gccgtcggac gcgaactttt gttcgttact aatcctgtgg
     1201 tggtgaacgg gtccgtcctg gtccctgagg atcaatgtta ctgtgccgga tggattgaat
     1261 ctcgcggcac gaacggcgct cagaccgcgt caaatgtcct gcagtggctt gcagcaggat
     1321 tcagcatttt gctgctgatg ttctatgcct accaaacctg gaaatctaca tgcggctggg
     1381 aggagatcta tgtgtgcgcc attgaaatgg ttaaggtgat tctcgagttc ttttttgagt
     1441 ttaagaatcc ctctatgctc taccttgcca caggacaccg ggtgcagtgg ctgcgctatg
     1501 cagagtggct gctcacttgt cctgtcatcc ttatccgcct gagcaacctc accggcctga
     1561 gcaacgacta cagcaggaga accatgggac tccttgtctc agacatcggg actatcgtgt
     1621 ggggggctac cagcgccatg gcaaccggct atgttaaagt catcttcttt tgtcttggat
     1681 tgtgctatgg cgcgaacaca ttttttcacg ccgccaaagc atatatcgag ggttatcata
     1741 ctgtgccaaa gggtcggtgc cgccaggtcg tgaccggcat ggcatggctg tttttcgtga
     1801 gctggggtat gttcccaatt ctcttcattt tggggcccga aggttttggc gtcctgagcg
     1861 tctatggctc caccgtaggt cacacgatta ttgatctgat gagtaaaaat tgttgggggt
     1921 tgttgggaca ctacctgcgc gtcctgatcc acgagcacat attgattcac ggagatatcc
     1981 gcaaaaccac caaactgaac atcggcggaa cggagatcga ggtcgagact ctcgtcgaag
     2041 acgaagccga ggccggagcc gtgccagcgg ccgccgtgag caagggcgag gaggataaca
     2101 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     2161 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     2221 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     2281 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     2341 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     2401 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     2461 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     2521 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     2581 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     2641 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     2701 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     2761 gcatggacga gctgtacaag taaggcgcgc cccacccctg cagggaattc gatatcaagc
     2821 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2881 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2941 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3001 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3061 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3121 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3181 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3241 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3301 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3361 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3421 gataccgagc gctgctcgag agatctgtga tagcggccat caagctggcc gcgactctag
     3481 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3541 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3601 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3661 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcagctta tcgataccgc
     3721 atgcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3781 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3841 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3901 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3961 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4021 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4081 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4141 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     4201 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4261 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4321 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4381 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4441 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4501 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4561 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4621 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4681 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4741 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4801 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4861 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4921 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4981 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5041 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5101 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5161 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5221 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5281 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5341 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5401 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5461 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5521 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5581 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5641 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5701 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5821 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5881 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5941 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6001 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6061 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6121 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6181 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6241 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6301 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6361 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6421 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6481 tgt
//
