LOCUS       sequence-421881-        6164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing GCaMP6f driven by SST interneuron-targeting
            enhancer. Alias: pAAV_WDS0004_GCaMP6f.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6164)
  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 6164)
  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_GCaMP6f
FEATURES             Location/Qualifiers
     source          1..6164
                     /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      1111..1120
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1117..1125,1126..1143,1144..1146,1147..1179,1180..1182,
                     1183..1206,1207..1233,1234..1290,1291..1296,1297..1566,
                     1567..1584,1585..2019,2020..2025,2026..2469)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1213..1222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1503..1524
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1630..1651)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1891..1910)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2510..3098
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2563..2583)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2981..2992)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3267..3348
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3304..3323)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3358..3377
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3427..3567
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3427..3556
                     /label=AAV2 ITR
     rep_origin      3642..4097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3729..3748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3939..3960
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4114..4133)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4233..4255
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4293..4311)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4379..4483
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4484..4552,4553..5344)
                     /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(4702..4721)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5515..6103
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6004..6023
                     /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 agcgcccggt gccaccatgg gttctcatca tcatcatcat
     1141 catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac
     1201 gataaggatc tcgccaccat ggtcgactca tcacgtcgta agtggaataa gacaggtcac
     1261 gcagtcagag ctataggtcg gctgagctca ctcgagaacg tctatatcaa ggccgacaag
     1321 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcggcgtg
     1381 cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1441 gacaaccact acctgagcgt gcagtccaaa ctttcgaaag accccaacga gaagcgcgat
     1501 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1561 tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt caccggggtg
     1621 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1681 gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1741 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1801 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1861 tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1921 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1981 gaggacggca acatcctggg gcacaagctg gagtacaacc tgccggacca actgactgaa
     2041 gagcagatcg cagaatttaa agaggaattc tccctatttg acaaggacgg ggatgggaca
     2101 ataacaacca aggagctggg gacggtgatg cggtctctgg ggcagaaccc cacagaagca
     2161 gagctgcagg acatgatcaa tgaagtagat gccgacggtg acggcacaat cgacttccct
     2221 gagttcctga caatgatggc aagaaaaatg aaatacaggg acacggaaga agaaattaga
     2281 gaagcgttcg gtgtgtttga taaggatggc aatggctaca tcagtgcagc agagcttcgc
     2341 cacgtgatga caaaccttgg agagaagtta acagatgaag aggttgatga aatgatcagg
     2401 gaagcagaca tcgatgggga tggtcaggta aactacgaag agtttgtaca aatgatgaca
     2461 gcgaagtgag gcgcgcccct gcagggaatt cgatatcaag cttatcgata atcaacctct
     2521 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2581 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2641 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2701 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2761 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2821 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2881 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     2941 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3001 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3061 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     3121 gagatctgtg atagcggcca tcaagctggc cgcgactcta gatcataatc agccatacca
     3181 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3241 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3301 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3361 gtttgtccaa actcatcaat gtatcagctt atcgataccg catgcacgtg cggaccgagc
     3421 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3481 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3541 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3601 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3661 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     3721 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3781 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3841 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3901 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3961 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     4021 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4081 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4141 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4201 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4261 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4321 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4381 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4441 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4501 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4561 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4621 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4681 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4741 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4801 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4861 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4921 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4981 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5041 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     5101 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5161 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5221 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5281 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5341 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5401 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5461 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5521 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5581 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5641 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5701 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5761 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5821 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5881 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5941 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6001 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6061 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6121 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
