LOCUS       sequence-421869-        6464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing mCherry-tagged channelrhodopsin-2 driven by
            SST interneuron-targeting enhancer. Alias: 
            pAAV_WDS0004_ChR2_mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6464)
  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 6464)
  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_BiSSTe4_ChR2_mCherry
FEATURES             Location/Qualifiers
     source          1..6464
                     /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          958..1090
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1112..1121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1118..2044
                     /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             2057..2764
                     /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(2199..2217)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2467..2486
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2810..3398
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2863..2883)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3281..3292)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3567..3648
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3604..3623)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3658..3677
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3727..3867
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3727..3856
                     /label=AAV2 ITR
     rep_origin      3942..4397
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4029..4048)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4239..4260
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4414..4433)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4533..4555
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4593..4611)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4679..4783
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4784..4852,4853..5644)
                     /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(5002..5021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5815..6403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6304..6323
                     /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 tttcagcata agcaagatgg aatggacaat
      181 ctcttatttc atcctaagtt ttacaggagg tccttgtttt atgtaaattt ccaaccattt
      241 caaaagctct ttatgtgtgt attttttttt taaccatttt tgacagtttc aactctttct
      301 tctcccatga tttctttttg ctctcaactg ttatgggaga gacaccaggt tctgtatgca
      361 tttctgcttt gggttgaaaa ttttatgcac ttcaaatcaa gcttcattca catagctaaa
      421 ttcactgagg atacttcttc cctttgaaaa tttgacaaga ttcttaacaa cccttccccg
      481 ttctcctcat atttgttgag tttcctagtg catgtccttc tccctgcccc cagaataact
      541 agcaggtgtg acacaggcat gcattttgca ttttccaagt ctgtgatgct aactttaggc
      601 ctagaaacat tagcagattt cctcttttta tcatgcatac atttgctgct tctgttatcc
      661 aatctagtca ctttaaagaa ttttcttatt tcttttggga cctgtgcaat ctgttgcaag
      721 ctagaaatgt ccatggctag accaataaat tcactagata aatttacctg tggccacatt
      781 atttaaaagt ctgtgatcga tgtttaaact tcctgcagcc cgggctgggc ataaaagtca
      841 gggcagagcc atctattgct tacatttgct tctagcctgc aagtcgagga gcgcagcctt
      901 ccagaagcag agcgcggcgc cttaagctgc agaagttggt cgtgaggcac tgggcacgta
      961 agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag
     1021 agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt
     1081 ctctccacag cgagctcgct agcgcgatcg cgccaccatg gactatggcg gcgctttgtc
     1141 tgccgtcgga cgcgaacttt tgttcgttac taatcctgtg gtggtgaacg ggtccgtcct
     1201 ggtccctgag gatcaatgtt actgtgccgg atggattgaa tctcgcggca cgaacggcgc
     1261 tcagaccgcg tcaaatgtcc tgcagtggct tgcagcagga ttcagcattt tgctgctgat
     1321 gttctatgcc taccaaacct ggaaatctac atgcggctgg gaggagatct atgtgtgcgc
     1381 cattgaaatg gttaaggtga ttctcgagtt cttttttgag tttaagaatc cctctatgct
     1441 ctaccttgcc acaggacacc gggtgcagtg gctgcgctat gcagagtggc tgctcacttg
     1501 tcctgtcatc cttatccgcc tgagcaacct caccggcctg agcaacgact acagcaggag
     1561 aaccatggga ctccttgtct cagacatcgg gactatcgtg tggggggcta ccagcgccat
     1621 ggcaaccggc tatgttaaag tcatcttctt ttgtcttgga ttgtgctatg gcgcgaacac
     1681 attttttcac gccgccaaag catatatcga gggttatcat actgtgccaa agggtcggtg
     1741 ccgccaggtc gtgaccggca tggcatggct gtttttcgtg agctggggta tgttcccaat
     1801 tctcttcatt ttggggcccg aaggttttgg cgtcctgagc gtctatggct ccaccgtagg
     1861 tcacacgatt attgatctga tgagtaaaaa ttgttggggg ttgttgggac actacctgcg
     1921 cgtcctgatc cacgagcaca tattgattca cggagatatc cgcaaaacca ccaaactgaa
     1981 catcggcgga acggagatcg aggtcgagac tctcgtcgaa gacgaagccg aggccggagc
     2041 cgtgccagcg gccgccgtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     2101 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     2161 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     2221 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     2281 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     2341 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     2401 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     2461 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     2521 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     2581 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     2641 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     2701 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     2761 gtaaggcgcg ccccacccct gcagggaatt cgatatcaag cttatcgata atcaacctct
     2821 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2881 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2941 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3001 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3061 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3121 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3181 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     3241 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3301 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3361 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     3421 gagatctgtg atagcggcca tcaagctggc cgcgactcta gatcataatc agccatacca
     3481 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3541 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3601 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3661 gtttgtccaa actcatcaat gtatcagctt atcgataccg catgcacgtg cggaccgagc
     3721 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3781 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3841 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3901 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3961 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     4021 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4081 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4141 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4201 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4261 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4321 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4381 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4441 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4501 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4561 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4621 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4681 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4741 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4801 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4861 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4921 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4981 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5041 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5101 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5161 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5221 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5281 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5341 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5401 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5461 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5521 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5581 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5641 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5701 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5761 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5821 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5881 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5941 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6001 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6061 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6121 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6181 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6241 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6301 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6361 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6421 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
