LOCUS       40924_3214        6312 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 6312)
  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 6312)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6312)
  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..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(65..82)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(236..824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(998..1855)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1856..1960)
                     /label=AmpR promoter
     rep_origin      complement(1986..2441)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     repeat_region   2622..2751
                     /label=AAV2 ITR
     promoter        2827..3274
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3303..3312
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3309..3371
                     /product="leader sequence from mouse immunoglobulin kappa
                     light chain"
                     /label=Ig-kappa leader
     primer_bind     4415..4436
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4539..4560)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5067..5096
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5100..5246
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    5262..5850
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(5853..5869)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(5899..6020)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   6182..6311
                     /label=AAV2 ITR
ORIGIN
        1 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
       61 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      121 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      181 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      241 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      301 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      361 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      421 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      481 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      541 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      601 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      661 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      721 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      781 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      841 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      901 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      961 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1021 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1081 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     1141 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1201 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1261 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1321 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1381 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1441 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1501 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1561 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1621 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1681 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1741 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1801 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1861 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1921 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1981 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2041 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2101 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2161 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2221 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2281 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2341 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2401 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     2461 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc
     2521 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc
     2581 ggcctcagtg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct cgctcgctca
     2641 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     2701 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
     2761 tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc cagggccctg
     2821 cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac
     2881 gaccggcccc gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct
     2941 atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc
     3001 accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga
     3061 aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg
     3121 cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca
     3181 cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg
     3241 cagcggagga gtcgtgtcgt gcctgagagc gcagtccaat tcaagctgct agcaaggatc
     3301 ccgccaccat ggagacagac acactcctgc tatgggtact gctgctctgg gttccaggtt
     3361 ccactggtga cagatctgaa agcatcaatt tcgtgagctg gggcggtagc acccaggatg
     3421 cgcagaagca ggcctgggcc gacccgttca gcaaggccag cggcattacc gtggtccagg
     3481 atgggcccac cgactacggc aaactcaagg ccatggtcga aagcggcaac gtgcagtggg
     3541 acgtggtgga tgtagaggcc gacttcgcct tgcgcgccgc cgccgaaggc ctgctcgaac
     3601 ccctggattt ctcggtgatc cagcgcgaca agatcgaccc gcgcttcgtc tccgaccatg
     3661 gcgtgggctc gttcctgggt tctttcgtgc tcggctacaa cgaaggcaag ctcggagcca
     3721 gcaagcccca ggactggacc gcgctgttcg acaccaagac ctaccccggc aaacgcgccc
     3781 tctacaaatg gccgagccct ggcgtgctcg aactggccct gctcgccgac ggcgtaccgg
     3841 ccgacaagct ctacccgctg gacctggacc gcgccttcaa gaaactcgac accatcaaga
     3901 aagacatcgt ctggtggggc ggcggtgcac agtcgcagca gctgctggcc tccggcgaag
     3961 tcagcatggg ccagttctgg aacggtcgca tccacgccct gcaagaagac ggcgcgccag
     4021 tgggcgtgag ctggaagcag aacctggtga tggccgacat cctcgtcgtg cctaaaggca
     4081 cgaagaacaa agccgccgcg atgaagttcc tggccagtgc cagcagcgcc aaaggccagg
     4141 acgacttttc cgccctgacc gcctatgctc cagtgaacat cgacagtgtg cagcgcctcg
     4201 accttgcgca ggtccggatc accgccgaca agcagaagaa cggcatcatg gcgaacttca
     4261 agatccgcca caacgtggag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
     4321 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
     4381 tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     4441 ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatga
     4501 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     4561 taaacggcca caagttcagc gtgcgcggcg agggcgaggg cgatgccacc aacggcaagc
     4621 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     4681 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     4741 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc agcttcaagg
     4801 acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     4861 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     4921 agtacaactg gaacgccaac ctggcaccga acctgccgac cgcctacgtc aaggatcaga
     4981 tcaccctcga tttcgcctac tgggccaaga acggtccggc cattgcgaca cggtggaatg
     5041 aatggctagt caaactgcag gtcgacgaac aaaaactcat ctcagaagag gatctgaatg
     5101 ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc tttaaggtgg
     5161 tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt atcatcctca
     5221 tcatgctttg gcagaagaag ccacgttaga agcttatcga taatcaacct ctggattaca
     5281 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5341 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5401 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5461 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5521 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5581 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5641 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     5701 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5761 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5821 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga cccgggcggc
     5881 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     5941 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     6001 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     6061 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata
     6121 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     6181 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     6241 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     6301 cgagcgcgca gc
//
