LOCUS       sequence-211213-        7028 bp    DNA     circular SYN 14-APR-2025
DEFINITION  GABA Sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7028)
  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 7028)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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.CAG-FLEX.iGABASnFR.F102Y.Y137L
FEATURES             Location/Qualifiers
     source          1..7028
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1171..1204
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1275..1308
                     /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      1333..1342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1339..1401
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     primer_bind     2445..2466
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2569..2590)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3097..3126
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3130..3276
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(3304..3337)
                     /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(3400..3433)
                     /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(3468..3487)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3549..3571)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3851..4128)
                     /label=chicken beta-actin promoter
     repeat_region   4441..4570
                     /label=AAV2 ITR
     rep_origin      4642..5097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4729..4748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4939..4960
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5123..5227
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5228..5296,5297..6088)
                     /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(5446..5465)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6259..6847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6748..6767
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct tctcgagtgg atcatttgca gaattggccc
     1141 ttttcagtgt ttcgaacctc ttcgagggac ataacttcgt ataatgtatg ctatacgaag
     1201 ttatgggtcg atggtgagat ttggactaga gggtcgatgg tgatgcttgg atcaaaatag
     1261 gaagaatccc aatgataact tcgtataaag tatcctatac gaagttatac tagtggtacc
     1321 accggtgata tcgccaccat ggagacagac acactcctgc tatgggtact gctgctctgg
     1381 gttccaggtt ccactggtga cagatctgaa agcatcaatt tcgtgagctg gggcggtagc
     1441 acccaggatg cgcagaagca ggcctgggcc gacccgttca gcaaggccag cggcattacc
     1501 gtggtccagg atgggcccac cgactacggc aaactcaagg ccatggtcga aagcggcaac
     1561 gtgcagtggg acgtggtgga tgtagaggcc gacttcgcct tgcgcgccgc cgccgaaggc
     1621 ctgctcgaac ccctggattt ctcggtgatc cagcgcgaca agatcgaccc gcgcttcgtc
     1681 tccgaccatg gcgtgggctc gttcctgtat tctttcgtgc tcggctacaa cgaaggcaag
     1741 ctcggagcca gcaagcccca ggactggacc gcgctgttcg acaccaagac ctaccccggc
     1801 aaacgcgccc tcctaaaatg gccgagccct ggcgtgctcg aactggccct gctcgccgac
     1861 ggcgtaccgg ccgacaagct ctacccgctg gacctggacc gcgccttcaa gaaactcgac
     1921 accatcaaga aagacatcgt ctggtggggc ggcggtgcac agtcgcagca gctgctggcc
     1981 tccggcgaag tcagcatggg ccagttctgg aacggtcgca tccacgccct gcaagaagac
     2041 ggcgcgccag tgggcgtgag ctggaagcag aacctggtga tggccgacat cctcgtcgtg
     2101 cctaaaggca cgaagaacaa agccgccgcg atgaagttcc tggccagtgc cagcagcgcc
     2161 aaaggccagg acgacttttc cgccctgacc gcctatgctc cagtgaacat cgacagtgtg
     2221 cagcgcctcg accttgcgca ggtccggatc accgccgaca agcagaagaa cggcatcatg
     2281 gcgaacttca agatccgcca caacgtggag gacggcagcg tgcagctcgc cgaccactac
     2341 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2401 acccagtccg tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2461 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga
     2521 gggagcatga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     2581 gacggcgacg taaacggcca caagttcagc gtgcgcggcg agggcgaggg cgatgccacc
     2641 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     2701 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     2761 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     2821 agcttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2881 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     2941 cacaagctgg agtacaactg gaacgccaac ctggcaccga acctgccgac cgcctacgtc
     3001 aaggatcaga tcaccctcga tttcgcctac tgggccaaga acggtccggc cattgcgaca
     3061 cggtggaatg aatggctagt caaactgcag gtcgacgaac aaaaactcat ctcagaagag
     3121 gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc
     3181 tttaaggtgg tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt
     3241 atcatcctca tcatgctttg gcagaagaag ccacgttagg atatcggatc cgctagcact
     3301 agtataactt cgtatagcat acattatacg aagttatggg tcgatggtga tgcttggcaa
     3361 ttcgggtcga tggtgaagca ttggtcttcc tattttgata taacttcgta taggatactt
     3421 tatacgaagt tatggggatc caattctttg ccaaaatgat gagacagcac aataaccagc
     3481 acgttgccca ggagctgtag gaaaaagaag aaggcatgaa catggttagc agaggctcta
     3541 gagccgccgg tcacacgcca gaagccgaac cccgccctgc cccgtccccc ccgaaggcag
     3601 ccgtcccccc gcggacagcc ccgagccgct ccccccgcac aaagggccct cccggagccc
     3661 ctcaaggctt tcacgcagcc acagaaaaga aacaagccgt cattaaacca agcgctaatt
     3721 acagcccgga ggagaagggc cgtcccgccc gctcacctgt gggagtaacg cggtcagtca
     3781 gagccggggc gggcggcgcg aggcggcggc ggagcggggc acggggcgaa ggcagcgtcg
     3841 cagcgactcc cgcccgccgc gcgcttcgct ttttataggg ccgccgccgc cgccgcctcg
     3901 ccataaaagg aaactttcgg agcgcgccgc tctgattggc tgccgccgca cctctccgcc
     3961 tcgccccgcc ccgcccctcg ccccgccccg ccccgcctgg cgcgcgcccc cccccccccc
     4021 ccgcccccat cgctgcacaa aataattaaa aaataaataa atacaaaatt gggggtgggg
     4081 agggggggga gatggggaga gtgaagcaga acgtggggct cacctcgacc atggtaatag
     4141 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     4201 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     4261 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     4321 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatggta
     4381 cgatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
     4441 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4501 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4561 gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     4621 gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4681 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4741 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4801 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4861 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4921 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4981 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     5041 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     5101 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5161 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5221 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5281 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5341 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5401 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5461 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5521 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5581 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5641 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5701 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5761 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5821 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5881 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5941 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6001 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6061 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6121 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6181 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6241 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6301 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6361 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6421 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6481 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6541 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6601 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6661 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6721 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6781 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6841 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6901 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6961 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     7021 agcggaag
//
