LOCUS       sequence-213878-        7845 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 7845)
  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 7845)
  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.Syn-FLEX.iGABASnFR.F102Y.Y137L.mRuby3
FEATURES             Location/Qualifiers
     source          1..7845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2515..2537
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2529..2546
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2530..2546
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2856..2977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2893..2912)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2947..2966
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3055..3643
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3161..3172
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3570..3590
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3715..3748
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3819..3852
                     /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      3877..3886
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3883..3942
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     4989..5010
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5113..5134)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5641..5670
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5674..5820
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(6592..6625)
                     /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(6688..6721)
                     /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(6732..6751)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          6734..6866
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   7459..7588
                     /label=AAV2 ITR
     primer_bind     complement(7639..7661)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7650..7666
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7674..7680,7681..7698,7699..7704))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7719..7740
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tctctgcgcg ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggmrwtgsgt aacgccaggg ttttcccagt
     2521 cacgacgttg taaaacgacg gccagtgaat tgtaataata cgasctrcgc gmbcgcccgt
     2581 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2641 tcagtgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt
     2701 taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag atccttatcg
     2761 attttaccac atttgtagag gttttacttg ctttaaaaaa cctcccacat ctccccctga
     2821 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2881 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2941 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgctcg aagcggccgc
     3001 ccgggtcgac tctagatgca tgctcgagcg gccgcccggg tcgacggtat cgatgcgggg
     3061 aggcggccca aagggagatc cgactcgtct gagggcgaag gcgaagacgc ggaagaggcc
     3121 gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg attgagggcc gaagggacgt
     3181 agcagaagga cgtcccgcgc agaatccagg tggcaacaca ggcgagcagc caaggaaagg
     3241 acgatgattt ccccgacaac accacggaat tgtcagtgcc caacagccga gcccctgtcc
     3301 agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aatagggagg gggaaagcga
     3361 aagtcccgga aaggagctga caggtggtgg caatgcccca accagtgggg gttgcgtcag
     3421 caaacacagt gcacaccacg ccacgttgcc tgacaacggg ccacaactcc tcataaagag
     3481 acagcaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
     3541 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
     3601 taccagtcaa tctttcacaa attttgtaat ccagaggttg attatcgata agcttctcga
     3661 gtggatcatt tgcagaattg gcccttttca gtgtttcgaa cctcttcgag ggacataact
     3721 tcgtataatg tatgctatac gaagttatgg gtcgatggtg agatttggac tagagggtcg
     3781 atggtgatgc ttggatcaaa ataggaagaa tcccaatgat aacttcgtat aaagtatcct
     3841 atacgaagtt atactagtgg taccaccggt gatatcgcca ccatggagac agacacactc
     3901 ctgctatggg tactgctgct ctgggttcca ggttccactg gtgacagatc tgaaagcatc
     3961 aatttcgtga gctggggcgg tagcacccag gatgcgcaga agcaggcctg ggccgacccg
     4021 ttcagcaagg ccagcggcat taccgtggtc caggatgggc ccaccgacta cggcaaactc
     4081 aaggccatgg tcgaaagcgg caacgtgcag tgggacgtgg tggatgtaga ggccgacttc
     4141 gccttgcgcg ccgccgccga aggcctgctc gaacccctgg atttctcggt gatccagcgc
     4201 gacaagatcg acccgcgctt cgtctccgac catggcgtgg gctcgttcct gtattctttc
     4261 gtgctcggct acaacgaagg caagctcgga gccagcaagc cccaggactg gaccgcgctg
     4321 ttcgacacca agacctaccc cggcaaacgc gccctcctaa aatggccgag ccctggcgtg
     4381 ctcgaactgg ccctgctcgc cgacggcgta ccggccgaca agctctaccc gctggacctg
     4441 gaccgcgcct tcaagaaact cgacaccatc aagaaagaca tcgtctggtg gggcggcggt
     4501 gcacagtcgc agcagctgct ggcctccggc gaagtcagca tgggccagtt ctggaacggt
     4561 cgcatccacg ccctgcaaga agacggcgcg ccagtgggcg tgagctggaa gcagaacctg
     4621 gtgatggccg acatcctcgt cgtgcctaaa ggcacgaaga acaaagccgc cgcgatgaag
     4681 ttcctggcca gtgccagcag cgccaaaggc caggacgact tttccgccct gaccgcctat
     4741 gctccagtga acatcgacag tgtgcagcgc ctcgaccttg cgcaggtccg gatcaccgcc
     4801 gacaagcaga agaacggcat catggcgaac ttcaagatcc gccacaacgt ggaggacggc
     4861 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     4921 ctgcccgaca accactacct gagcacccag tccgtgctga gcaaagaccc caacgagaag
     4981 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     5041 gagctgtaca agggcggtac cggagggagc atgagcaagg gcgaggagct gttcaccggg
     5101 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgcgc
     5161 ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat ctgcaccacc
     5221 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     5281 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     5341 ggctacgtcc aggagcgcac catcagcttc aaggacgacg gcacctacaa gacccgcgcc
     5401 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     5461 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actggaacgc caacctggca
     5521 ccgaacctgc cgaccgccta cgtcaaggat cagatcaccc tcgatttcgc ctactgggcc
     5581 aagaacggtc cggccattgc gacacggtgg aatgaatggc tagtcaaact gcaggtcgac
     5641 gaacaaaaac tcatctcaga agaggatctg aatgctgtgg gccaggacac gcaggaggtc
     5701 atcgtggtgc cacactcctt gccctttaag gtggtggtga tctcagccat cctggccctg
     5761 gtggtgctca ccatcatctc ccttatcatc ctcatcatgc tttggcagaa gaagccacgt
     5821 tctagaggct ccggaaatgc tgtgggccag gacacgcagg agcgtatggt gtctaagggc
     5881 gaagagctga tcaaggaaaa tatgcgtatg aaggtggtca tggaaggttc ggtcaacggc
     5941 caccaattca aatgcacagg tgaaggagaa ggcaggccgt acgagggagt gcaaaccatg
     6001 aggatcaaag tcatcgaggg aggacccctg ccatttgcct ttgacattct tgccacgtcg
     6061 ttcatgtatg gcagccgtac ttttatcaag tacccggcag acattcctga tttctttaaa
     6121 cagtcctttc ctgagggttt tacttgggaa agagttacga gatacgaaga tggtggagtc
     6181 gtcaccgtca cgcaggacac cagccttgag gatggcgaac tcgtttacaa cgtcaaagtc
     6241 agaggggtaa actttccctc caatggtccc gtgatgcaga agaagaccaa gggttgggag
     6301 cctaatacag agatgatgta tccagcagat ggtggtctga ggggatacac tgatattgca
     6361 ctgaaagttg atggtggtgg ccatctgcat tgcaatttcg taacaactta caggtcaaaa
     6421 aagaccgtcg ggaacatcaa gatgcccggt gtccatgccg ttgatcaccg cctggaaagg
     6481 atagaggaaa gtgacaatga aacgtacgta gtacaacgcg aagtcgcagt tgccaagtac
     6541 tccaatcttg gtggtggtat ggatgaactt tataagtaag ctagcactag tataacttcg
     6601 tatagcatac attatacgaa gttatgtgtc gatggtgatg cttggcaatt cgggtcgatg
     6661 gtgaagcatt ggtcttccta ttttgatata acttcgtata ggatacttta tacgaagtta
     6721 tggggatcat cacctgtgga gagaaaggca aagtggatgt cagtaagacc aataggtgcc
     6781 tatcagaaac gcaagagtct tctctgtctc gacaagccca gtttctattg gtctccttaa
     6841 acctgtcttg taaccttgat acttacctga tccaacggct gcgacttggg gcagggggtc
     6901 ctaggggtgg tctggccagg agccgcgggg gcggactgcg cggtgcccag gagcacagct
     6961 gcgctctcag gcacgacacg actcctccgc tgcccaccgc agactgaggc agcgctgagt
     7021 cgccggcgcc gcagcgcaga tggtcgcgcc cgtgcccccc tatctcgcgc ctcgcgtggt
     7081 gcggtccggc tgggccggcg gcggcgcgga cgcgaccaag gtggccggga aggggagttt
     7141 gcgggggacc ggcgagtgac gtcagcgcgc cttcagtgct gaggcggcgg tggcgcgcgc
     7201 cgccaggcgg gggcgaaggc actgtccgcg gtgctgaagc tggcagtgcg cacgcgcctc
     7261 gccgcatcct gtttcccctc cccctctctg ataggggatg cgcaatttgg ggaatggggg
     7321 ttgggtgctt gtccagtggg tcggggtcgg tcgtcaattt aaatactgaa aagggcgaat
     7381 tccagcacac tggagctcgg atcttcctag agcatggcta cgtagataag tagcatggcg
     7441 ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     7501 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     7561 cggcctcagt gagcgagcga gcgcgcagct ggcgtaatag cgaagaggcc cgcaccgatc
     7621 gcccttccct agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     7681 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     7741 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7801 tgaatcagcc aaagggaacc cctagtgatg gagttggcca ctccc
//
