LOCUS       sequence-213447-        7554 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 7554)
  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 7554)
  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: pMito.iGABASnFR.mRuby3
FEATURES             Location/Qualifiers
     source          1..7554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        389..592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     543..563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     637..656
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        637..655
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             736..810
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     primer_bind     1887..1908
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2011..2032)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3304..3321)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3310..3534
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3666..4254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3746..3765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4583..4630
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4655..4674
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4862..5656)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASKPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4973..4992)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5583..5602
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5755..5774)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        5853..6044
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6001..6021
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(6104..6895,6896..6964))
                     /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     6727..6746
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6965..7069)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7096..7551
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7183..7202)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7393..7414
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgcgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      421 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      481 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      541 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagaa ctctctggct
      601 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
      661 ccaagcttgg taccgaactc ggatccacta gtaacggccg ccagtgtgct ggaattcggc
      721 ttggggatat ccaccatgag cgtgctgacc cccctgctgc tgaggggcct gaccggcagc
      781 gccaggaggc tgcccgtgcc cagggccaag atccacagcc tgggcgaccc ccccgtggcc
      841 accagatctg aaagcatcaa tttcgtgagc tggggcggta gcacccagga tgcgcagaag
      901 caggcctggg ccgacccgtt cagcaaggcc agcggcatta ccgtggtcca ggatgggccc
      961 accgactacg gcaaactcaa ggccatggtc gaaagcggca acgtgcagtg ggacgtggtg
     1021 gatgtagagg ccgacttcgc cttgcgcgcc gccgccgaag gcctgctcga acccctggat
     1081 ttctcggtga tccagcgcga caagatcgac ccgcgcttcg tctccgacca tggcgtgggc
     1141 tcgttcctgt tctctttcgt gctcggctac aacgaaggca agctcggagc cagcaagccc
     1201 caggactgga ccgcgctgtt cgacaccaag acctaccccg gcaaacgcgc cctctacaaa
     1261 tggccgagcc ctggcgtgct cgaactggcc ctgctcgccg acggcgtacc ggccgacaag
     1321 ctctacccgc tggacctgga ccgcgccttc aagaaactcg acaccatcaa gaaagacatc
     1381 gtctggtggg gcggcggtgc acagtcgcag cagctgctgg cctccggcga agtcagcatg
     1441 ggccagttct ggaacggtcg catccacgcc ctgcaagaag acggcgcgcc agtgggcgtg
     1501 agctggaagc agaacctggt gatggccgac atcctcgtcg tgcctaaagg cacgaagaac
     1561 aaagccgccg cgatgaagtt cctggccagt gccagcagcg ccaaaggcca ggacgacttt
     1621 tccgccctga ccgcctatgc tccagtgaac atcgacagtg tgcagcgcct cgaccttgcg
     1681 caggtccgga tcaccgccga caagcagaag aacggcatca tggcgaactt caagatccgc
     1741 cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1801 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgtgctgagc
     1861 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1921 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat gagcaagggc
     1981 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2041 cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg
     2101 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2161 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2221 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcagcttcaa ggacgacggc
     2281 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2341 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2401 tggaacgcca acctggcacc gaacctgccg accgcctacg tcaaggatca gatcaccctc
     2461 gatttcgcct actgggccaa gaacggtccg gccattgcga cacggtggaa tgaatggcta
     2521 gtcaaactgc agggctccgg aaatgctgtg ggccaggaca cgcaggagcg tatggtgtct
     2581 aagggcgaag agctgatcaa ggaaaatatg cgtatgaagg tggtcatgga aggttcggtc
     2641 aacggccacc aattcaaatg cacaggtgaa ggagaaggca ggccgtacga gggagtgcaa
     2701 accatgagga tcaaagtcat cgagggagga cccctgccat ttgcctttga cattcttgcc
     2761 acgtcgttca tgtatggcag ccgtactttt atcaagtacc cggcagacat tcctgatttc
     2821 tttaaacagt cctttcctga gggttttact tgggaaagag ttacgagata cgaagatggt
     2881 ggagtcgtca ccgtcacgca ggacaccagc cttgaggatg gcgaactcgt ttacaacgtc
     2941 aaagtcagag gggtaaactt tccctccaat ggtcccgtga tgcagaagaa gaccaagggt
     3001 tgggagccta atacagagat gatgtatcca gcagatggtg gtctgagggg atacactgat
     3061 attgcactga aagttgatgg tggtggccat ctgcattgca atttcgtaac aacttacagg
     3121 tcaaaaaaga ccgtcgggaa catcaagatg cccggtgtcc atgccgttga tcaccgcctg
     3181 gaaaggatag aggaaagtga caatgaaacg tacgtagtac aacgcgaagt cgcagttgcc
     3241 aagtactcca atcttggtgg tggtatggat gaactttata agtaggcggc cgctcgagat
     3301 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3361 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3421 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3481 gggaggattg ggaagacaat agcaggcgtg ctggggatgc ggtgggctct atggcttctg
     3541 aggcggaaag aaccagtggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3601 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3661 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3721 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3781 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3841 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3901 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3961 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4021 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4081 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4141 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4201 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4261 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4321 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4381 tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg
     4441 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa
     4501 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
     4561 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt
     4621 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     4681 ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag
     4741 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg
     4801 aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg
     4861 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     4921 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     4981 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     5041 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
     5101 cgacgagatc ctcgccgtcg ggcttgctcg ccttgagcct ggcgaacagt tcggctggcg
     5161 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     5221 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     5281 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     5341 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     5401 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     5461 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     5521 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     5581 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     5641 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag
     5701 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgggcctcgg
     5761 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     5821 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     5881 gcttgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     5941 actaattgag atgcttgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     6001 accctaactg acacacattc cacagctggt tctttccgcc tcaggactct tcctttttca
     6061 ataaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6121 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6181 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6241 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6301 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6361 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6421 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6481 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6541 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6601 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6661 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6721 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6781 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6841 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6901 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6961 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7021 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7081 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     7141 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     7201 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     7261 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     7321 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     7381 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     7441 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     7501 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tacg
//
