LOCUS       sequence-421878-        6183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV construct expressing GCaMP6f driven by PV+ basket cell-targeting
            enhancer. Alias: pAAV_WDP0003_GCaMP6f.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6183)
  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 6183)
  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_GCaMP6f
FEATURES             Location/Qualifiers
     source          1..6183
                     /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      1130..1139
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1136..1144,1145..1162,1163..1165,1166..1198,1199..1201,
                     1202..1225,1226..1252,1253..1309,1310..1315,1316..1585,
                     1586..1603,1604..2038,2039..2044,2045..2488)
                     /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      1232..1241
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1522..1543
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1649..1670)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1910..1929)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2529..3117
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2582..2602)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3000..3011)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3286..3367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3323..3342)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3377..3396
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3446..3586
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3446..3575
                     /label=AAV2 ITR
     rep_origin      3661..4116
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3748..3767)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3958..3979
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4133..4152)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4252..4274
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4312..4330)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4398..4502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4503..4571,4572..5363)
                     /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(4721..4740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5534..6122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6023..6042
                     /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 gcgcccggtg ccaccatggg
     1141 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
     1201 ggatctgtac gacgatgacg ataaggatct cgccaccatg gtcgactcat cacgtcgtaa
     1261 gtggaataag acaggtcacg cagtcagagc tataggtcgg ctgagctcac tcgagaacgt
     1321 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttcaaga tccgccacaa
     1381 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     1441 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga
     1501 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1561 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gcaagggcga
     1621 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1681 caagttcagc gtgtccggcg agggtgaggg cgatgccacc tacggcaagc tgaccctgaa
     1741 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1801 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1861 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa
     1921 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1981 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacct
     2041 gccggaccaa ctgactgaag agcagatcgc agaatttaaa gaggaattct ccctatttga
     2101 caaggacggg gatgggacaa taacaaccaa ggagctgggg acggtgatgc ggtctctggg
     2161 gcagaacccc acagaagcag agctgcagga catgatcaat gaagtagatg ccgacggtga
     2221 cggcacaatc gacttccctg agttcctgac aatgatggca agaaaaatga aatacaggga
     2281 cacggaagaa gaaattagag aagcgttcgg tgtgtttgat aaggatggca atggctacat
     2341 cagtgcagca gagcttcgcc acgtgatgac aaaccttgga gagaagttaa cagatgaaga
     2401 ggttgatgaa atgatcaggg aagcagacat cgatggggat ggtcaggtaa actacgaaga
     2461 gtttgtacaa atgatgacag cgaagtgagg cgcgcccctg cagggaattc gatatcaagc
     2521 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2581 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2641 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2701 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2761 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2821 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2881 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     2941 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3001 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3061 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3121 gataccgagc gctgctcgag agatctgtga tagcggccat caagctggcc gcgactctag
     3181 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3241 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3301 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3361 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcagctta tcgataccgc
     3421 atgcacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3481 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3541 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3601 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3661 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3721 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3781 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3841 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3901 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3961 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     4021 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     4081 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4141 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4201 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4261 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4321 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4381 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4441 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4501 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4561 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4621 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4681 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4741 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4801 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4861 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4921 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4981 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5041 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5101 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5161 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5221 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5281 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5341 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5401 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5461 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5521 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5581 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5641 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5701 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5761 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5821 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5881 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5941 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6001 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6061 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6121 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6181 tgt
//
