LOCUS       sequence-421852-        6344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing mCherry-tagged channelrhodopsin-2 driven by
            chandelier cell-targeting enhancer. Alias: 
            pAAV_WDC0004_ChR2_mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6344)
  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 6344)
  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_BiPVe4_ChR2_mCherry
FEATURES             Location/Qualifiers
     source          1..6344
                     /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          838..970
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      992..1001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             998..1924
                     /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             1937..2644
                     /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(2079..2097)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2347..2366
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2690..3278
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2743..2763)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3161..3172)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3447..3528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3484..3503)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3538..3557
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3607..3747
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3607..3736
                     /label=AAV2 ITR
     rep_origin      3822..4277
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3909..3928)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4119..4140
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4294..4313)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4413..4435
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4473..4491)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4559..4663
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4664..4732,4733..5524)
                     /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(4882..4901)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5695..6283
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6184..6203
                     /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 ggtgtatttc ctgtgtgggc taatgtagct
      181 aactttgttt ggactccagg tttctgaggc agtttcaaat tcttgattta tgggttcatt
      241 tttggatcat ggcagcctcc ctgctgaaac ccccatagtt tcccttcaat aaggaaggct
      301 ggatgagatc acagctgagg aatcctggtg gtgtgtactg tgttatttct ctcctagtgc
      361 tttgcctaca cccttccatc atgcttacat ttggacctga aaataattgg ttgaagctgc
      421 tcctcacttt tagcctttta tccgagtttg acagtacatc accccaaata acacggacta
      481 taaatggctt gaacctaatg agtgctcttt gtggacaagc cgtccattct ccttttcttg
      541 agagggaaat taccggctcc taagcttcta tttccagttc tatagaatgt gggcgctaat
      601 ttgaaagcag cctcataaaa tggaagcaaa caggctaatt agaaatattt atgggcctgt
      661 tagtgaatat atgccatcga tgtttaaact tcctgcagcc cgggctgggc ataaaagtca
      721 gggcagagcc atctattgct tacatttgct tctagcctgc aagtcgagga gcgcagcctt
      781 ccagaagcag agcgcggcgc cttaagctgc agaagttggt cgtgaggcac tgggcacgta
      841 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
      901 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
      961 ctctccacag cgagctcgct agcgcgatcg cgccaccatg gactatggcg gcgctttgtc
     1021 tgccgtcgga cgcgaacttt tgttcgttac taatcctgtg gtggtgaacg ggtccgtcct
     1081 ggtccctgag gatcaatgtt actgtgccgg atggattgaa tctcgcggca cgaacggcgc
     1141 tcagaccgcg tcaaatgtcc tgcagtggct tgcagcagga ttcagcattt tgctgctgat
     1201 gttctatgcc taccaaacct ggaaatctac atgcggctgg gaggagatct atgtgtgcgc
     1261 cattgaaatg gttaaggtga ttctcgagtt cttttttgag tttaagaatc cctctatgct
     1321 ctaccttgcc acaggacacc gggtgcagtg gctgcgctat gcagagtggc tgctcacttg
     1381 tcctgtcatc cttatccgcc tgagcaacct caccggcctg agcaacgact acagcaggag
     1441 aaccatggga ctccttgtct cagacatcgg gactatcgtg tggggggcta ccagcgccat
     1501 ggcaaccggc tatgttaaag tcatcttctt ttgtcttgga ttgtgctatg gcgcgaacac
     1561 attttttcac gccgccaaag catatatcga gggttatcat actgtgccaa agggtcggtg
     1621 ccgccaggtc gtgaccggca tggcatggct gtttttcgtg agctggggta tgttcccaat
     1681 tctcttcatt ttggggcccg aaggttttgg cgtcctgagc gtctatggct ccaccgtagg
     1741 tcacacgatt attgatctga tgagtaaaaa ttgttggggg ttgttgggac actacctgcg
     1801 cgtcctgatc cacgagcaca tattgattca cggagatatc cgcaaaacca ccaaactgaa
     1861 catcggcgga acggagatcg aggtcgagac tctcgtcgaa gacgaagccg aggccggagc
     1921 cgtgccagcg gccgccgtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     1981 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     2041 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     2101 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     2161 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     2221 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     2281 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     2341 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     2401 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     2461 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     2521 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     2581 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     2641 gtaaggcgcg ccccacccct gcagggaatt cgatatcaag cttatcgata atcaacctct
     2701 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2761 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2821 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2881 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2941 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3001 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3061 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     3121 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3181 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3241 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     3301 gagatctgtg atagcggcca tcaagctggc cgcgactcta gatcataatc agccatacca
     3361 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3421 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3481 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3541 gtttgtccaa actcatcaat gtatcagctt atcgataccg catgcacgtg cggaccgagc
     3601 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3661 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3721 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3781 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3841 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     3901 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3961 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4021 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4081 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4141 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4201 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4261 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4321 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4381 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4441 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4501 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4561 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4621 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4681 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4741 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4801 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4861 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4921 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4981 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5041 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5101 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5161 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5221 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5281 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5341 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5401 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5461 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5521 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5581 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5641 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5701 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5761 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5821 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5881 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5941 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6001 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6061 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6121 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6181 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6241 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6301 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
