LOCUS       sequence-421853-        6044 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing GCaMP6f driven by chandelier cell-targeting
            enhancer. Alias: pAAV_WDC0004_GCaMP6f.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6044)
  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 6044)
  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_GCaMP6f
FEATURES             Location/Qualifiers
     source          1..6044
                     /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      991..1000
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(997..1005,1006..1023,1024..1026,1027..1059,1060..1062,
                     1063..1086,1087..1113,1114..1170,1171..1176,1177..1446,
                     1447..1464,1465..1899,1900..1905,1906..2349)
                     /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      1093..1102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1383..1404
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1510..1531)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1771..1790)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2390..2978
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2443..2463)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2861..2872)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3147..3228
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3184..3203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3238..3257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3307..3447
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3307..3436
                     /label=AAV2 ITR
     rep_origin      3522..3977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3609..3628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3819..3840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3994..4013)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4113..4135
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4173..4191)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4259..4363
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4364..4432,4433..5224)
                     /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(4582..4601)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5395..5983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5884..5903
                     /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 agcgcccggt gccaccatgg gttctcatca tcatcatcat
     1021 catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac
     1081 gataaggatc tcgccaccat ggtcgactca tcacgtcgta agtggaataa gacaggtcac
     1141 gcagtcagag ctataggtcg gctgagctca ctcgagaacg tctatatcaa ggccgacaag
     1201 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcggcgtg
     1261 cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1321 gacaaccact acctgagcgt gcagtccaaa ctttcgaaag accccaacga gaagcgcgat
     1381 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1441 tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt caccggggtg
     1501 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1561 gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1621 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1681 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1741 tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1801 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1861 gaggacggca acatcctggg gcacaagctg gagtacaacc tgccggacca actgactgaa
     1921 gagcagatcg cagaatttaa agaggaattc tccctatttg acaaggacgg ggatgggaca
     1981 ataacaacca aggagctggg gacggtgatg cggtctctgg ggcagaaccc cacagaagca
     2041 gagctgcagg acatgatcaa tgaagtagat gccgacggtg acggcacaat cgacttccct
     2101 gagttcctga caatgatggc aagaaaaatg aaatacaggg acacggaaga agaaattaga
     2161 gaagcgttcg gtgtgtttga taaggatggc aatggctaca tcagtgcagc agagcttcgc
     2221 cacgtgatga caaaccttgg agagaagtta acagatgaag aggttgatga aatgatcagg
     2281 gaagcagaca tcgatgggga tggtcaggta aactacgaag agtttgtaca aatgatgaca
     2341 gcgaagtgag gcgcgcccct gcagggaatt cgatatcaag cttatcgata atcaacctct
     2401 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2461 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2521 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2581 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2641 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2701 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2761 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     2821 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2881 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2941 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     3001 gagatctgtg atagcggcca tcaagctggc cgcgactcta gatcataatc agccatacca
     3061 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3121 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3181 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3241 gtttgtccaa actcatcaat gtatcagctt atcgataccg catgcacgtg cggaccgagc
     3301 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3361 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3421 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     3481 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3541 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     3601 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3661 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3721 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3781 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3841 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3901 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3961 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4021 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4081 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4141 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     4201 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4261 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4321 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4381 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4441 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4501 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4561 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4621 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4681 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4741 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4801 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4861 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4921 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4981 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5041 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5101 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5161 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5221 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5281 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5341 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5401 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5461 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5521 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5581 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5641 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5701 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5761 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5821 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5881 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5941 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6001 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt
//
