LOCUS       sequence-270142-        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
FEATURES             Location/Qualifiers
     source          1..6771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(530..1118)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(610..629)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1289..2080,2081..2149))
                     /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     1912..1931
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2150..2254)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2280..2735)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2417..2438)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2629..2648
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2807..2936
                     /label=AAV2 ITR
     intron          3529..3661
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3644..3663
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    3674..3707
                     /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    3770..3803
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3831..3977)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(3981..4010)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     4517..4538
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4641..4662)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(5709..5768)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      5765..5774
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5799..5832)
                     /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(5903..5936)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6008..6596
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6061..6081)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6479..6490)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    join(6674..6771,1..24)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6685..6704)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6739..6758
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
       61 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
      121 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
      181 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
      241 tgaggacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt
      301 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      361 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      421 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      481 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      541 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      601 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      661 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      721 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
      781 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      841 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      901 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      961 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1021 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1081 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1141 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1201 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1261 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1321 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1381 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1441 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1501 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1561 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1621 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1681 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1741 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1801 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1861 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1921 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1981 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2041 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2101 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2161 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2221 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     2281 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2341 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2401 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2461 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2521 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2581 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2641 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2701 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     2761 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgc gcgctcgctc
     2821 gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc
     2881 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta
     2941 gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctagga agatccgagc
     3001 tccagtgtgc tggaattcgc ccttttcagt atttaaattg acgaccgacc ccgacccact
     3061 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg
     3121 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc
     3181 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact
     3241 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg
     3301 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg
     3361 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc
     3421 gtgcctgaga gcgcagctgt gctcctgggc accgcgcagt ccgcccccgc ggctcctggc
     3481 cagaccaccc ctaggacccc ctgccccaag tcgcagccgt tggatcaggt aagtatcaag
     3541 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
     3601 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     3661 ggtgatgatc cccataactt cgtataaagt atcctatacg aagttatatc aaaataggaa
     3721 gaccaatgct tcaccatcga cccgaattgc caagcatcac catcgacaca taacttcgta
     3781 taatgtatgc tatacgaagt tatactagtg ctagcggatc cgatatccta acgtggcttc
     3841 ttctgccaaa gcatgatgag gatgataagg gagatgatgg tgagcaccac cagggccagg
     3901 atggctgaga tcaccaccac cttaaagggc aaggagtgtg gcaccacgat gacctcctgc
     3961 gtgtcctggc ccacagcatt cagatcctct tctgagatga gtttttgttc gtcgacctgc
     4021 agtttgacta gccattcatt ccaccgtgtc gcaatggccg gaccgttctt ggcccagtag
     4081 gcgaaatcga gggtgatctg atccttgacg taggcggtcg gcaggttcgg tgccaggttg
     4141 gcgttccagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     4201 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     4261 ttgtaggtgc cgtcgtcctt gaagctgatg gtgcgctcct ggacgtagcc ttcgggcatg
     4321 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     4381 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     4441 atgaacttca gggtcagctt gccgttggtg gcatcgccct cgccctcgcc gcgcacgctg
     4501 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     4561 agctcctcgc ccttgctcat gctccctccg gtaccgccct tgtacagctc gtccatgccg
     4621 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     4681 gggtctttgc tcagcacgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg
     4741 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcc
     4801 acgttgtggc ggatcttgaa gttcgccatg atgccgttct tctgcttgtc ggcggtgatc
     4861 cggacctgcg caaggtcgag gcgctgcaca ctgtcgatgt tcactggagc ataggcggtc
     4921 agggcggaaa agtcgtcctg gcctttggcg ctgctggcac tggccaggaa cttcatcgcg
     4981 gcggctttgt tcttcgtgcc tttaggcacg acgaggatgt cggccatcac caggttctgc
     5041 ttccagctca cgcccactgg cgcgccgtct tcttgcaggg cgtggatgcg accgttccag
     5101 aactggccca tgctgacttc gccggaggcc agcagctgct gcgactgtgc accgccgccc
     5161 caccagacga tgtctttctt gatggtgtcg agtttcttga aggcgcggtc caggtccagc
     5221 gggtagagct tgtcggccgg tacgccgtcg gcgagcaggg ccagttcgag cacgccaggg
     5281 ctcggccatt tgtagagggc gcgtttgccg gggtaggtct tggtgtcgaa cagcgcggtc
     5341 cagtcctggg gcttgctggc tccgagcttg ccttcgttgt agccgagcac gaaagagaac
     5401 aggaacgagc ccacgccatg gtcggagacg aagcgcgggt cgatcttgtc gcgctggatc
     5461 accgagaaat ccaggggttc gagcaggcct tcggcggcgg cgcgcaaggc gaagtcggcc
     5521 tctacatcca ccacgtccca ctgcacgttg ccgctttcga ccatggcctt gagtttgccg
     5581 tagtcggtgg gcccatcctg gaccacggta atgccgctgg ccttgctgaa cgggtcggcc
     5641 caggcctgct tctgcgcatc ctgggtgcta ccgccccagc tcacgaaatt gatgctttca
     5701 gatctgtcac cagtggaacc tggaacccag agcagcagta cccatagcag gagtgtgtct
     5761 gtctccatgg tggcgatatc accggtggta ccactagtat aacttcgtat aggatacttt
     5821 atacgaagtt atcattggga ttcttcctat tttgatccaa gcatcaccat cgaccctcta
     5881 gtccaaatct caccatcgac ccataacttc gtatagcata cattatacga agttatgtcc
     5941 ctcgaagagg ttcgaaacac tgaaaagggc caattctgca aatgatccac tcgagaagct
     6001 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     6061 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     6121 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     6181 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     6241 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     6301 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     6361 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     6421 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     6481 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     6541 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     6601 ataccgtcga cccgggcggc cgctcgagca tgcatctaga gtcgacccgg gcggccgctt
     6661 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     6721 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac c
//
