LOCUS       sequence-213866-        7849 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 7849)
  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 7849)
  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.F102G.mRuby3
FEATURES             Location/Qualifiers
     source          1..7849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      182..637
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(269..288)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     479..500
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        663..767
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(768..836,837..1628)
                     /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(986..1005)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1799..2387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2288..2307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2674..2695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2710..2715,2716..2733,2734..2740)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2748..2764
                     /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)."
     primer_bind     2753..2775
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2772..2788
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2772..2788
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3117..3238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3154..3173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3208..3227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3316..3904
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3422..3433
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3831..3851
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3976..4009
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4080..4113
                     /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      4138..4147
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     4144..4203
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     5250..5271
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5374..5395)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5902..5931
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5935..6081
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(6853..6886)
                     /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(6949..6982)
                     /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(6993..7012)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          6995..7127
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   7720..7849
                     /label=AAV2 ITR
ORIGIN
        1 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc ccacagtgac tgggaaaacc
       61 ctggcgttag gcwttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctggcgtaat
      121 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      181 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
      241 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      301 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      361 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      421 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      481 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      541 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      601 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
      661 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
      721 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
      781 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
      841 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
      901 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
      961 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1021 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1081 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1141 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1201 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1261 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1321 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1381 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1441 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1501 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1561 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1621 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     1681 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     1741 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1801 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1861 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1921 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1981 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2041 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2101 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2161 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2221 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2281 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2341 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2401 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     2461 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     2521 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     2581 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctgcgcg ctcgctcgct
     2641 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
     2701 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     2761 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcttaatt aacgccttaw
     2821 ttaggctgcg cgmtcgctcg ctcactgagg ccgcccgggc aaagcactga ggccgcccgg
     2881 gcaaagcccg ggcgtcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
     2941 tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat
     3001 gctctaggaa gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa
     3061 acctcccaca tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3121 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3181 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3241 atgtctgctc gaagcggccg cccgggtcga ctctagatgc atgctcgagc ggccgcccgg
     3301 gtcgacggta tcgatgcggg gaggcggccc aaagggagat ccgactcgtc tgagggcgaa
     3361 ggcgaagacg cggaagaggc cgcagagccg gcagcaggcc gcgggaagga aggtccgctg
     3421 gattgagggc cgaagggacg tagcagaagg acgtcccgcg cagaatccag gtggcaacac
     3481 aggcgagcag ccaaggaaag gacgatgatt tccccgacaa caccacggaa ttgtcagtgc
     3541 ccaacagccg agcccctgtc cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg
     3601 caatagggag ggggaaagcg aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc
     3661 aaccagtggg ggttgcgtca gcaaacacag tgcacaccac gccacgttgc ctgacaacgg
     3721 gccacaactc ctcataaaga gacagcaacc aggatttata caaggaggag aaaatgaaag
     3781 ccatacggga agcaatagca tgatacaaag gcattaaagc agcgtatcca catagcgtaa
     3841 aaggagcaac atagttaaga ataccagtca atctttcaca aattttgtaa tccagaggtt
     3901 gattatcgat aagcttctcg agtggatcat ttgcagaatt ggcccttttc agtgtttcga
     3961 acctcttcga gggacataac ttcgtataat gtatgctata cgaagttatg ggtcgatggt
     4021 gagatttgga ctagagggtc gatggtgatg cttggatcaa aataggaaga atcccaatga
     4081 taacttcgta taaagtatcc tatacgaagt tatactagtg gtaccaccgg tgatatcgcc
     4141 accatggaga cagacacact cctgctatgg gtactgctgc tctgggttcc aggttccact
     4201 ggtgacagat ctgaaagcat caatttcgtg agctggggcg gtagcaccca ggatgcgcag
     4261 aagcaggcct gggccgaccc gttcagcaag gccagcggca ttaccgtggt ccaggatggg
     4321 cccaccgact acggcaaact caaggccatg gtcgaaagcg gcaacgtgca gtgggacgtg
     4381 gtggatgtag aggccgactt cgccttgcgc gccgccgccg aaggcctgct cgaacccctg
     4441 gatttctcgg tgatccagcg cgacaagatc gacccgcgct tcgtctccga ccatggcgtg
     4501 ggctcgttcc tgggttcttt cgtgctcggc tacaacgaag gcaagctcgg agccagcaag
     4561 ccccaggact ggaccgcgct gttcgacacc aagacctacc ccggcaaacg cgccctctac
     4621 aaatggccga gccctggcgt gctcgaactg gccctgctcg ccgacggcgt accggccgac
     4681 aagctctacc cgctggacct ggaccgcgcc ttcaagaaac tcgacaccat caagaaagac
     4741 atcgtctggt ggggcggcgg tgcacagtcg cagcagctgc tggcctccgg cgaagtcagc
     4801 atgggccagt tctggaacgg tcgcatccac gccctgcaag aagacggcgc gccagtgggc
     4861 gtgagctgga agcagaacct ggtgatggcc gacatcctcg tcgtgcctaa aggcacgaag
     4921 aacaaagccg ccgcgatgaa gttcctggcc agtgccagca gcgccaaagg ccaggacgac
     4981 ttttccgccc tgaccgccta tgctccagtg aacatcgaca gtgtgcagcg cctcgacctt
     5041 gcgcaggtcc ggatcaccgc cgacaagcag aagaacggca tcatggcgaa cttcaagatc
     5101 cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     5161 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgtgctg
     5221 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     5281 gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag catgagcaag
     5341 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     5401 ggccacaagt tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc
     5461 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     5521 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     5581 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcagctt caaggacgac
     5641 ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     5701 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     5761 aactggaacg ccaacctggc accgaacctg ccgaccgcct acgtcaagga tcagatcacc
     5821 ctcgatttcg cctactgggc caagaacggt ccggccattg cgacacggtg gaatgaatgg
     5881 ctagtcaaac tgcaggtcga cgaacaaaaa ctcatctcag aagaggatct gaatgctgtg
     5941 ggccaggaca cgcaggaggt catcgtggtg ccacactcct tgccctttaa ggtggtggtg
     6001 atctcagcca tcctggccct ggtggtgctc accatcatct cccttatcat cctcatcatg
     6061 ctttggcaga agaagccacg ttctagaggc tccggaaatg ctgtgggcca ggacacgcag
     6121 gagcgtatgg tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc
     6181 atggaaggtt cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcaggccg
     6241 tacgagggag tgcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc
     6301 tttgacattc ttgccacgtc gttcatgtat ggcagccgta cttttatcaa gtacccggca
     6361 gacattcctg atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg
     6421 agatacgaag atggtggagt cgtcaccgtc acgcaggaca ccagccttga ggatggcgaa
     6481 ctcgtttaca acgtcaaagt cagaggggta aactttccct ccaatggtcc cgtgatgcag
     6541 aagaagacca agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg
     6601 aggggataca ctgatattgc actgaaagtt gatggtggtg gccatctgca ttgcaatttc
     6661 gtaacaactt acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tgtccatgcc
     6721 gttgatcacc gcctggaaag gatagaggaa agtgacaatg aaacgtacgt agtacaacgc
     6781 gaagtcgcag ttgccaagta ctccaatctt ggtggtggta tggatgaact ttataagtaa
     6841 gctagcacta gtataacttc gtatagcata cattatacga agttatgtgt cgatggtgat
     6901 gcttggcaat tcgggtcgat ggtgaagcat tggtcttcct attttgatat aacttcgtat
     6961 aggatacttt atacgaagtt atggggatca tcacctgtgg agagaaaggc aaagtggatg
     7021 tcagtaagac caataggtgc ctatcagaaa cgcaagagtc ttctctgtct cgacaagccc
     7081 agtttctatt ggtctcctta aacctgtctt gtaaccttga tacttacctg atccaacggc
     7141 tgcgacttgg ggcagggggt cctaggggtg gtctggccag gagccgcggg ggcggactgc
     7201 gcggtgccca ggagcacagc tgcgctctca ggcacgacac gactcctccg ctgcccaccg
     7261 cagactgagg cagcgctgag tcgccggcgc cgcagcgcag atggtcgcgc ccgtgccccc
     7321 ctatctcgcg cctcgcgtgg tgcggtccgg ctgggccggc ggcggcgcgg acgcgaccaa
     7381 ggtggccggg aaggggagtt tgcgggggac cggcgagtga cgtcagcgcg ccttcagtgc
     7441 tgaggcggcg gtggcgcgcg ccgccaggcg ggggcgaagg cactgtccgc ggtgctgaag
     7501 ctggcagtgc gcacgcgcct cgccgcatcc tgtttcccct ccccctctct gataggggat
     7561 gcgcaatttg gggaatgggg gttgggtgct tgtccagtgg gtcggggtcg gtcgtcaatt
     7621 taaatactga aaagggcgaa ttccagcaca ctggagctcg gatcttccta gagcatggct
     7681 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     7741 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     7801 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcag
//
