LOCUS       sequence-271175-        6139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector to drive the expression of GCaMP6f in PV cortical 
            interneuronsunder the control of the E2 regulatory element.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6139)
  AUTHORS   Vormstein-Schneider D, Lin JD, Pelkey KA, Chittajallu R, Guo B, 
            Arias-Garcia MA, Allaway K, Sakopoulos S, Schneider G, Stevenson O, 
            Vergara J, Sharma J, Zhang Q, Franken TP, Smith J, Ibrahim LA, M 
            Astro KJ, Sabri E, Huang S, Favuzzi E, Burbridge T, Xu Q, Guo L, 
            Vogel I, Sanchez V, Saldi GA, Gorissen BL, Yuan X, Zaghloul KA, 
            Devinsky O, Sabatini BL, Batista-Brito R, Reynolds J, Feng G, Fu Z, 
            McBain CJ, Fishell G, Dimidschstein J
  TITLE     Viral manipulation of functionally distinct interneurons in mice, 
            non-human primates and humans.
  JOURNAL   Nat Neurosci. 2020 Dec;23(12):1629-1636. doi: 
            10.1038/s41593-020-0692-9. Epub 2020 Aug 17.
  PUBMED    32807948
REFERENCE   2  (bases 1 to 6139)
  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: 
            "10.1038/s41593-020-0692-9"; journalName: "Nat Neurosci"; date: 
            "2020-12"; volume: "23"; issue: "12"; pages: "1629-1636"
COMMENT     Sequence Label: pAAV-S5E2-GCaMP6f
FEATURES             Location/Qualifiers
     source          1..6139
                     /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          934..1066
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     regulatory      1086..1095
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1092..1100,1101..1118,1119..1121,1122..1154,1155..1157,
                     1158..1181,1182..1208,1209..1265,1266..1271,1272..1541,
                     1542..1559,1560..1994,1995..2000,2001..2444)
                     /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      1188..1197
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1478..1499
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1605..1626)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1866..1885)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2485..3073
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2538..2558)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2956..2967)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3242..3323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3279..3298)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3333..3352
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   3402..3542
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3402..3531
                     /label=AAV2 ITR
     rep_origin      3617..4072
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3704..3723)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3914..3935
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4089..4108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4208..4230
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4268..4286)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4354..4458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4459..4527,4528..5319)
                     /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(4677..4696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5490..6078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5979..5998
                     /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 gcgtttaatt aaactagtgg tttaatctaa catggctgct
      181 atagcttact gactagaagt taagtgcaca cttcctaaaa gaaggctttg acacaagcca
      241 cttcagttcc ctcctcattt tcttgtcccc attcctctct ctgtagaatt ctgagatttc
      301 aattcagttt tatacagaaa ccacattact gtaagcccta caaagttatg gcaatatagc
      361 tatatggagt caagtaatgt aggttatttt tttcccaatg gtgctggtga aggtggcaat
      421 tatgtagcta tacttagcag actgaggaaa ttctgctaga gtcagcattt gtctcttcat
      481 tgctatgaaa cagtaatgga aaaataaaca aaaacaaaag gcaaacacta tgcataattc
      541 cctcagatca tattaacatg tgatgttgga gtaaattgtt ataaccccat tttggaaata
      601 cttaccttaa ttaactatga tttccttaaa ataatgcagt atttacaatc tatatgaaag
      661 cactatatgg gacacatggt atgatggaac agtgcaccca agagacacca agaacattcc
      721 tgtctgtggc agtcttttct ctatacagag gcatttagtc tcaattgctc agagttattt
      781 taaacttcct gcagcccggg ctgggcataa aagtcagggc agagccatct attgcttaca
      841 tttgcttcta gcctgcaagt cgaggagcgc agccttccag aagcagagcg cggcgcctta
      901 agctgcagaa gttggtcgtg aggcactggg cacgtaagta tcaaggttac aagacaggtt
      961 taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag
     1021 gcacctattg gtcttactga catccacttt gcctttctct ccacagcgag ctcgctagta
     1081 ccggtgccac catgggttct catcatcatc atcatcatgg tatggctagc atgactggtg
     1141 gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatctcgcc accatggtcg
     1201 actcatcacg tcgtaagtgg aataagacag gtcacgcagt cagagctata ggtcggctga
     1261 gctcactcga gaacgtctat atcaaggccg acaagcagaa gaacggcatc aaggcgaact
     1321 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga
     1381 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcgtgcagt
     1441 ccaaactttc gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     1501 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggagca
     1561 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1621 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg tgagggcgat gccacctacg
     1681 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1741 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1801 agcacgactt cttcaagtcc gccatgcccg aaggctacat ccaggagcgc accatcttct
     1861 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1921 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1981 agctggagta caacctgccg gaccaactga ctgaagagca gatcgcagaa tttaaagagg
     2041 aattctccct atttgacaag gacggggatg ggacaataac aaccaaggag ctggggacgg
     2101 tgatgcggtc tctggggcag aaccccacag aagcagagct gcaggacatg atcaatgaag
     2161 tagatgccga cggtgacggc acaatcgact tccctgagtt cctgacaatg atggcaagaa
     2221 aaatgaaata cagggacacg gaagaagaaa ttagagaagc gttcggtgtg tttgataagg
     2281 atggcaatgg ctacatcagt gcagcagagc ttcgccacgt gatgacaaac cttggagaga
     2341 agttaacaga tgaagaggtt gatgaaatga tcagggaagc agacatcgat ggggatggtc
     2401 aggtaaacta cgaagagttt gtacaaatga tgacagcgaa gtgaggcgcg cccctgcagg
     2461 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2521 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2581 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2641 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2701 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2761 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2821 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2881 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     2941 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     3001 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     3061 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctgtgatagc ggccatcaag
     3121 ctggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt
     3181 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt
     3241 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3301 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3361 agcttatcga taccgcatgc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     3421 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3481 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     3541 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     3601 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3661 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     3721 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3781 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     3841 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3901 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     3961 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     4021 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     4081 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4141 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4201 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4261 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4321 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4381 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4441 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4501 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4561 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4621 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4681 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4741 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4801 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4861 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4921 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4981 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5041 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5101 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5161 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5221 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5281 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5341 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5401 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5461 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5521 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5581 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5641 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5701 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5761 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5821 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5881 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5941 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6001 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6061 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6121 tggccttttg ctcacatgt
//
