LOCUS       sequence-213667-        7025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GABA Sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7025)
  AUTHORS   Marvin JS, Shimoda Y, Magloire V, Leite M, Kawashima T, Jensen TP, 
            Kolb I, Knott EL, Novak O, Podgorski K, Leidenheimer NJ, Rusakov DA,
            Ahrens MB, Kullmann DM, Looger LL
  TITLE     A genetically encoded fluorescent sensor for in vivo imaging of 
            GABA.
  JOURNAL   Nat Methods. 2019 Aug;16(8):763-770. doi: 10.1038/s41592-019-0471-2.
            Epub 2019 Jul 15.
  PUBMED    31308547
REFERENCE   2  (bases 1 to 7025)
  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/s41592-019-0471-2"; journalName: "Nat Methods"; date: 
            "2019-08"; volume: "16"; issue: "8"; pages: "763-770"
COMMENT     Sequence Label: pAAV.CAG-FLEX.iGABASnFR
FEATURES             Location/Qualifiers
     source          1..7025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1171..1204
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1275..1308
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      1333..1342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1339..1398
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     2445..2466
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2569..2590)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3097..3126
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3130..3276
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(3304..3337)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3400..3433)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(3468..3487)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3549..3571)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3850..4127)
                     /label=chicken beta-actin promoter
     repeat_region   4438..4567
                     /label=AAV2 ITR
     rep_origin      4639..5094
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4726..4745)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4936..4957
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5120..5224
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5225..5293,5294..6085)
                     /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(5443..5462)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6256..6844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6745..6764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct tctcgagtgg atcatttgca gaattggccc
     1141 ttttcagtgt ttcgaacctc ttcgagggac ataacttcgt ataatgtatg ctatacgaag
     1201 ttatgggtcg atggtgagat ttggactaga gggtcgatgg tgatgcttgg atcaaaatag
     1261 gaagaatccc aatgataact tcgtataaag tatcctatac gaagttatac tagtggtacc
     1321 accggtgata tcgccaccat ggagacagac acactcctgc tatgggtact gctgctctgg
     1381 gttccaggtt ccactggtga cagatctgaa agcatcaatt tcgtgagctg gggcggtagc
     1441 acccaggatg cgcagaagca ggcctgggcc gacccgttca gcaaggccag cggcattacc
     1501 gtggtccagg atgggcccac cgactacggc aaactcaagg ccatggtcga aagcggcaac
     1561 gtgcagtggg acgtggtgga tgtagaggcc gacttcgcct tgcgcgccgc cgccgaaggc
     1621 ctgctcgaac ccctggattt ctcggtgatc cagcgcgaca agatcgaccc gcgcttcgtc
     1681 tccgaccatg gcgtgggctc gttcctgttc tctttcgtgc tcggctacaa cgaaggcaag
     1741 ctcggagcca gcaagcccca ggactggacc gcgctgttcg acaccaagac ctaccccggc
     1801 aaacgcgccc tctacaaatg gccgagccct ggcgtgctcg aactggccct gctcgccgac
     1861 ggcgtaccgg ccgacaagct ctacccgctg gacctggacc gcgccttcaa gaaactcgac
     1921 accatcaaga aagacatcgt ctggtggggc ggcggtgcac agtcgcagca gctgctggcc
     1981 tccggcgaag tcagcatggg ccagttctgg aacggtcgca tccacgccct gcaagaagac
     2041 ggcgcgccag tgggcgtgag ctggaagcag aacctggtga tggccgacat cctcgtcgtg
     2101 cctaaaggca cgaagaacaa agccgccgcg atgaagttcc tggccagtgc cagcagcgcc
     2161 aaaggccagg acgacttttc cgccctgacc gcctatgctc cagtgaacat cgacagtgtg
     2221 cagcgcctcg accttgcgca ggtccggatc accgccgaca agcagaagaa cggcatcatg
     2281 gcgaacttca agatccgcca caacgtggag gacggcagcg tgcagctcgc cgaccactac
     2341 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2401 acccagtccg tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2461 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga
     2521 gggagcatga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2581 gacggcgacg taaacggcca caagttcagc gtgcgcggcg agggcgaggg cgatgccacc
     2641 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2701 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2761 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2821 agcttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2881 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2941 cacaagctgg agtacaactg gaacgccaac ctggcaccga acctgccgac cgcctacgtc
     3001 aaggatcaga tcaccctcga tttcgcctac tgggccaaga acggtccggc cattgcgaca
     3061 cggtggaatg aatggctagt caaactgcag gtcgacgaac aaaaactcat ctcagaagag
     3121 gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc
     3181 tttaaggtgg tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt
     3241 atcatcctca tcatgctttg gcagaagaag ccacgttagg atatcggatc cgctagcact
     3301 agtataactt cgtatagcat acattatacg aagttatggg tcgatggtga tgcttggcaa
     3361 ttcgggtcga tggtgaagca ttggtcttcc tattttgata taacttcgta taggatactt
     3421 tatacgaagt tatggggatc caattctttg ccaaaatgat gagacagcac aataaccagc
     3481 acgttgccca ggagctgtag gaaaaagaag aaggcatgaa catggttagc agaggctcta
     3541 gagccgccgg tcacacgcca gaagccgaac cccgccctgc cccgtccccc ccgaaggcag
     3601 ccgtcccccc gcggacagcc ccgagccgct ccccccgcac aaagggccct cccggagccc
     3661 ctcaaggctt tcacgcagcc acagaaaaga aacaagccgt cattaaacca agcgctaatt
     3721 acagcccgga ggagaagggc cgtcccgccc gctcacctgt gggagtaacg cggtcagtca
     3781 gagccggggc gggcggcgcg aggcggcggc ggagcggggc acggggcgaa ggcagcgcgc
     3841 agcgactccc gcccgccgcg cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc
     3901 cataaaagga aactttcgga gcgcgccgct ctgattggct gccgccgcac ctctccgcct
     3961 cgccccgccc cgccccccgc cccgccccgc cccccctggc gcgcgccccc cccccccccc
     4021 cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga
     4081 ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc
     4141 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     4201 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     4261 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     4321 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatggtac
     4381 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg
     4441 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     4501 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4561 cgcgcagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     4621 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4681 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4741 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4801 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4861 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4921 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4981 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     5041 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     5101 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5161 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5221 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5281 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5341 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5401 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5461 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5521 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5581 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5641 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5701 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5761 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5821 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5881 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5941 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6001 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6061 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6121 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6181 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6241 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6301 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6361 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6421 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6481 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6541 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6601 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6661 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6721 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6781 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6841 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6901 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6961 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7021 ggaag
//
