LOCUS Exported 7028 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION GABA Sensor. ACCESSION . VERSION . KEYWORDS pAAV.CAG-FLEX.iGABASnFR.F102G 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7028 /organism="synthetic DNA construct" /mol_type="other DNA" 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 complement(1171..1204) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(1275..1308) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites (Lee and Saito, 1988)." regulatory 1333..1342 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" sig_peptide 1339..1401 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader 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 3304..3337 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 3400..3433 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). 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 5228..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" primer_bind 7001..7018 /label=L4440 /note="L4440 vector, 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 gttcctgggc tctttcgtgc tcggctacaa cgaaggcaag 1741 ctcggagcca gcaagcccca ggactggacc gcgctgttcg acaccaagac ctaccccggc 1801 aaacgcgccc tctacaaatg 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 //