LOCUS       40924_3219        6302 bp DNA     circular SYN 13-MAY-2021
DEFINITION  GABA Sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6302)
  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 6302)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6302)
  AUTHORS   .
  TITLE     Direct Submission
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     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(66..83)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(999..1856)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1857..1961)
                     /label=AmpR promoter
     rep_origin      complement(1987..2442)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     repeat_region   2686..2815
                     /label=AAV2 ITR
     promoter        2891..3338
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3367..3376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3373..3435
                     /product="leader sequence from mouse immunoglobulin kappa
                     light chain"
                     /label=Ig-kappa leader
     primer_bind     4479..4500
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4603..4624)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5131..5160
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5164..5310
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    5326..5914
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(5917..5933)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(5963..6084)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1321 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1921 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1981 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2041 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2101 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2161 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2221 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2281 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2341 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2401 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     2461 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg
     2521 ctcgctcgct cactgaggcc gcgcgggagt agggtgacgc cgcggsgaac ccccacccca
     2581 cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac
     2641 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctgcg cgctcgctcg
     2701 ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg cccggcctca
     2761 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg ttccttgtag
     2821 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatccagggc
     2881 cctgcgtatg agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc
     2941 tgacgaccgg ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc
     3001 ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt
     3061 cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca
     3121 ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg
     3181 gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg
     3241 ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg
     3301 tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc caattcaagc tgctagcaag
     3361 gatcccgcca ccatggagac agacacactc ctgctatggg tactgctgct ctgggttcca
     3421 ggttccactg gtgacagatc tgaaagcatc aatttcgtga gctggggcgg tagcacccag
     3481 gatgcgcaga agcaggcctg ggccgacccg ttcagcaagg ccagcggcat taccgtggtc
     3541 caggatgggc ccaccgacta cggcaaactc aaggccatgg tcgaaagcgg caacgtgcag
     3601 tgggacgtgg tggatgtaga ggccgacttc gccttgcgcg ccgccgccga aggcctgctc
     3661 gaacccctgg atttctcggt gatccagcgc gacaagatcg acccgcgctt cgtctccgac
     3721 catggcgtgg gctcgttcct gttctctttc gtgctcggct acaacgaagg caagctcgga
     3781 gccagcaagc cccaggactg gaccgcgctg ttcgacacca agacctaccc cggcaaacgc
     3841 gccctctaca aatggccgag ccctggcgtg ctcgaactgg ccctgctcgc cgacggcgta
     3901 ccggccgaca agctctaccc gctggacctg gaccgcgcct tcaagaaact cgacaccatc
     3961 aagaaagaca tcgtctggtg gggcggcggt gcacagtcgc agcagctgct ggcctccggc
     4021 gaagtcagca tgggccagtt ctggaacggt cgcatccacg ccctgcaaga agacggcgcg
     4081 ccagtgggcg tgagctggaa gcagaacctg gtgatggccg acatcctcgt cgtgcctaaa
     4141 ggcacgaaga acaaagccgc cgcgatgaag ttcctggcca gtgccagcag cgccaaaggc
     4201 caggacgact tttccgccct gaccgcctat gctccagtga acatcgacag tgtgcagcgc
     4261 ctcgaccttg cgcaggtccg gatcaccgcc gacaagcaga agaacggcat catggcgaac
     4321 ttcaagatcc gccacaacgt ggaggacggc agcgtgcagc tcgccgacca ctaccagcag
     4381 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     4441 tccgtgctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     4501 accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc
     4561 atgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     4621 gacgtaaacg gccacaagtt cagcgtgcgc ggcgagggcg agggcgatgc caccaacggc
     4681 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     4741 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     4801 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcagcttc
     4861 aaggacgacg gcacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     4921 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     4981 ctggagtaca actggaacgc caacctggca ccgaacctgc cgaccgccta cgtcaaggat
     5041 cagatcaccc tcgatttcgc ctactgggcc aagaacggtc cggccattgc gacacggtgg
     5101 aatgaatggc tagtcaaact gcaggtcgac gaacaaaaac tcatctcaga agaggatctg
     5161 aatgctgtgg gccaggacac gcaggaggtc atcgtggtgc cacactcctt gccctttaag
     5221 gtggtggtga tctcagccat cctggccctg gtggtgctca ccatcatctc ccttatcatc
     5281 ctcatcatgc tttggcagaa gaagccacgt tagaagctta tcgataatca acctctggat
     5341 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5401 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5461 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5521 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     5581 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     5641 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     5701 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     5761 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     5821 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     5881 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgacccggg
     5941 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     6001 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     6061 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     6121 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     6181 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa
     6241 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gcgcgctcgc
     6301 ag
//
