LOCUS       sequence-458825-        6771 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 6771)
  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 6771)
  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.hSyn-FLEX.iGABASnFR.F102G
FEATURES             Location/Qualifiers
     source          1..6771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          723..855
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     838..857
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    868..901
                     /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)."
     protein_bind    964..997
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1025..1171)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1175..1204)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1711..1732
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1835..1856)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2903..2962)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2959..2968
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2993..3026)
                     /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)."
     protein_bind    complement(3097..3130)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3202..3790
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3255..3275)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3673..3684)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3868..3989
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3879..3898)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3933..3952
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4495..5083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4575..4594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5254..6045,6046..6114))
                     /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     5877..5896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6115..6219)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6245..6700)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6382..6403)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6594..6613
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatcc gagctccagt gtgctggaat tcgccctttt cagtatttaa attgacgacc
      241 gaccccgacc cactggacaa gcacccaacc cccattcccc aaattgcgca tcccctatca
      301 gagaggggga ggggaaacag gatgcggcga ggcgcgtgcg cactgccagc ttcagcaccg
      361 cggacagtgc cttcgccccc gcctggcggc gcgcgccacc gccgcctcag cactgaaggc
      421 gcgctgacgt cactcgccgg tcccccgcaa actccccttc ccggccacct tggtcgcgtc
      481 cgcgccgccg ccggcccagc cggaccgcac cacgcgaggc gcgagatagg ggggcacggg
      541 cgcgaccatc tgcgctgcgg cgccggcgac tcagcgctgc ctcagtctgc ggtgggcagc
      601 ggaggagtcg tgtcgtgcct gagagcgcag ctgtgctcct gggcaccgcg cagtccgccc
      661 ccgcggctcc tggccagacc acccctagga ccccctgccc caagtcgcag ccgttggatc
      721 aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga
      781 gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg
      841 cctttctctc cacaggtgat gatccccata acttcgtata aagtatccta tacgaagtta
      901 tatcaaaata ggaagaccaa tgcttcacca tcgacccgaa ttgccaagca tcaccatcga
      961 cacataactt cgtataatgt atgctatacg aagttatact agtgctagcg gatccgatat
     1021 cctaacgtgg cttcttctgc caaagcatga tgaggatgat aagggagatg atggtgagca
     1081 ccaccagggc caggatggct gagatcacca ccaccttaaa gggcaaggag tgtggcacca
     1141 cgatgacctc ctgcgtgtcc tggcccacag cattcagatc ctcttctgag atgagttttt
     1201 gttcgtcgac ctgcagtttg actagccatt cattccaccg tgtcgcaatg gccggaccgt
     1261 tcttggccca gtaggcgaaa tcgagggtga tctgatcctt gacgtaggcg gtcggcaggt
     1321 tcggtgccag gttggcgttc cagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     1381 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     1441 cctcggcgcg ggtcttgtag gtgccgtcgt ccttgaagct gatggtgcgc tcctggacgt
     1501 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc
     1561 tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     1621 tgccggtggt gcagatgaac ttcagggtca gcttgccgtt ggtggcatcg ccctcgccct
     1681 cgccgcgcac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     1741 ccaccccggt gaacagctcc tcgcccttgc tcatgctccc tccggtaccg cccttgtaca
     1801 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
     1861 cgcgcttctc gttggggtct ttgctcagca cggactgggt gctcaggtag tggttgtcgg
     1921 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     1981 cgctgccgtc ctccacgttg tggcggatct tgaagttcgc catgatgccg ttcttctgct
     2041 tgtcggcggt gatccggacc tgcgcaaggt cgaggcgctg cacactgtcg atgttcactg
     2101 gagcataggc ggtcagggcg gaaaagtcgt cctggccttt ggcgctgctg gcactggcca
     2161 ggaacttcat cgcggcggct ttgttcttcg tgcctttagg cacgacgagg atgtcggcca
     2221 tcaccaggtt ctgcttccag ctcacgccca ctggcgcgcc gtcttcttgc agggcgtgga
     2281 tgcgaccgtt ccagaactgg cccatgctga cttcgccgga ggccagcagc tgctgcgact
     2341 gtgcaccgcc gccccaccag acgatgtctt tcttgatggt gtcgagtttc ttgaaggcgc
     2401 ggtccaggtc cagcgggtag agcttgtcgg ccggtacgcc gtcggcgagc agggccagtt
     2461 cgagcacgcc agggctcggc catttgtaga gggcgcgttt gccggggtag gtcttggtgt
     2521 cgaacagcgc ggtccagtcc tggggcttgc tggctccgag cttgccttcg ttgtagccga
     2581 gcacgaaaga acccaggaac gagcccacgc catggtcgga gacgaagcgc gggtcgatct
     2641 tgtcgcgctg gatcaccgag aaatccaggg gttcgagcag gccttcggcg gcggcgcgca
     2701 aggcgaagtc ggcctctaca tccaccacgt cccactgcac gttgccgctt tcgaccatgg
     2761 ccttgagttt gccgtagtcg gtgggcccat cctggaccac ggtaatgccg ctggccttgc
     2821 tgaacgggtc ggcccaggcc tgcttctgcg catcctgggt gctaccgccc cagctcacga
     2881 aattgatgct ttcagatctg tcaccagtgg aacctggaac ccagagcagc agtacccata
     2941 gcaggagtgt gtctgtctcc atggtggcga tatcaccggt ggtaccacta gtataacttc
     3001 gtataggata ctttatacga agttatcatt gggattcttc ctattttgat ccaagcatca
     3061 ccatcgaccc tctagtccaa atctcaccat cgacccataa cttcgtatag catacattat
     3121 acgaagttat gtccctcgaa gaggttcgaa acactgaaaa gggccaattc tgcaaatgat
     3181 ccactcgaga agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     3241 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3301 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3361 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3421 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3481 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3541 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3601 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3661 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3721 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3781 gcctccccgc atcgataccg tcgacccggg cggccgctcg agcatgcatc tagagtcgac
     3841 ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3901 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3961 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     4021 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     4081 aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc
     4141 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     4201 ctcactgagg ccgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4261 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4321 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4381 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4441 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4681 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4921 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6241 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     6301 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6361 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     6421 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     6481 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     6541 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     6601 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     6661 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     6721 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca g
//
