LOCUS       Exported                6810 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  GABA Sensor.
ACCESSION   .
VERSION     .
KEYWORDS    pMito.iGABASnFR.R205A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6810)
  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 6810)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6810
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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(2560..2577)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2566..2790
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2922..3510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3002..3021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3839..3886
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3911..3930
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4118..4912)
                     /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(4229..4248)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4839..4858
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5011..5030)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5257..5277
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(5360..6220)
                     /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     5983..6002
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6221..6325)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6352..6807
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6439..6458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6649..6670
                     /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 ggaacggtgc 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 aggtcgacta ggcggccgct cgagatcagc ctcgactgtg ccttctagtt
     2581 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2641 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2701 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2761 ggcgtgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agtggcggta
     2821 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2881 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2941 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3001 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3061 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3121 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3181 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3241 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3301 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3361 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3421 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3481 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3541 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3601 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3661 gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct
     3721 tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa
     3781 tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga
     3841 acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt
     3901 ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac
     3961 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     4021 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     4081 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     4141 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     4201 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     4261 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     4321 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggct
     4381 tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     4441 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     4501 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     4561 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     4621 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     4681 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat
     4741 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     4801 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     4861 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     4921 atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa aaagcctcct
     4981 cactacttct ggaatagctc agaggccggg cctcggcctc tgcataaata aaaaaaatta
     5041 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     5101 ttaggggcgg gactatggtt gctgactaat tgagatgctt gctttgcata cttctgcctg
     5161 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc ttgctttgca
     5221 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
     5281 gctggttctt tccgcctcag gactcttcct ttttcaataa atcaatctaa agtatatatg
     5341 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     5401 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     5461 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5521 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     5581 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5641 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5701 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5761 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5821 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5881 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5941 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     6001 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     6061 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     6121 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6181 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     6241 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     6301 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     6361 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     6421 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     6481 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     6541 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     6601 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     6661 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     6721 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6781 ttaacaaaat attaacgctt acaatttacg
//
