LOCUS Exported 6771 bp ds-DNA circular SYN 05-JAN-2023 DEFINITION GABA Sensor. ACCESSION . VERSION . KEYWORDS pAAV.hSyn-FLEX.iGABASnFR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6771) 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 6771) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 5, 2023 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6771 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(359..376) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(530..1118) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(610..629) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1289..2149) /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 1912..1931 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2150..2254) /gene="bla" /label=AmpR promoter rep_origin complement(2280..2735) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2417..2438) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2629..2648 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 2807..2936 /label=AAV2 ITR intron 3529..3661 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 3644..3663 /label=pMT2-F /note="Synthetic intron, forward primer" protein_bind complement(3674..3707) /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)." protein_bind complement(3770..3803) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(3831..3977) /codon_start=1 /product="transmembrane domain from platelet derived growth factor receptor beta" /label=PDGFR-beta TM domain /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR" CDS complement(3981..4010) /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind 4517..4538 /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4641..4662) /label=EGFP-C /note="EGFP, forward primer" sig_peptide complement(5706..5768) /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader regulatory 5765..5774 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 5799..5832 /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)." protein_bind 5903..5936 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 6008..6596 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6061..6081) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6479..6490) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 6674..24 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6685..6704) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6739..6758 /label=SV40pA-R /note="SV40 polyA, reverse primer" ORIGIN 1 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt 61 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc 121 gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa 181 ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac 241 tgaggacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt 301 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 361 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 421 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 481 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 541 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 601 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 661 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 721 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 781 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 841 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 901 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 961 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 1021 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 1081 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 1141 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 1201 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 1261 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 1321 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 1381 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 1441 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 1501 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 1561 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 1621 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 1681 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 1741 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 1801 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 1861 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 1921 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 1981 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 2041 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 2101 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 2161 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 2221 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta 2281 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 2341 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat 2401 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 2461 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta 2521 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 2581 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 2641 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 2701 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa 2761 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgc gcgctcgctc 2821 gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc 2881 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta 2941 gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctagga agatccgagc 3001 tccagtgtgc tggaattcgc ccttttcagt atttaaattg acgaccgacc ccgacccact 3061 ggacaagcac ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg 3121 aaacaggatg cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc 3181 gcccccgcct ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact 3241 cgccggtccc ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg 3301 cccagccgga ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg 3361 ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc 3421 gtgcctgaga gcgcagctgt gctcctgggc accgcgcagt ccgcccccgc ggctcctggc 3481 cagaccaccc ctaggacccc ctgccccaag tcgcagccgt tggatcaggt aagtatcaag 3541 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc 3601 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca 3661 ggtgatgatc cccataactt cgtataaagt atcctatacg aagttatatc aaaataggaa 3721 gaccaatgct tcaccatcga cccgaattgc caagcatcac catcgacaca taacttcgta 3781 taatgtatgc tatacgaagt tatactagtg ctagcggatc cgatatccta acgtggcttc 3841 ttctgccaaa gcatgatgag gatgataagg gagatgatgg tgagcaccac cagggccagg 3901 atggctgaga tcaccaccac cttaaagggc aaggagtgtg gcaccacgat gacctcctgc 3961 gtgtcctggc ccacagcatt cagatcctct tctgagatga gtttttgttc gtcgacctgc 4021 agtttgacta gccattcatt ccaccgtgtc gcaatggccg gaccgttctt ggcccagtag 4081 gcgaaatcga gggtgatctg atccttgacg taggcggtcg gcaggttcgg tgccaggttg 4141 gcgttccagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg 4201 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc 4261 ttgtaggtgc cgtcgtcctt gaagctgatg gtgcgctcct ggacgtagcc ttcgggcatg 4321 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg 4381 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag 4441 atgaacttca gggtcagctt gccgttggtg gcatcgccct cgccctcgcc gcgcacgctg 4501 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac 4561 agctcctcgc ccttgctcat gctccctccg gtaccgccct tgtacagctc gtccatgccg 4621 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg 4681 gggtctttgc tcagcacgga ctgggtgctc aggtagtggt tgtcgggcag cagcacgggg 4741 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgct gccgtcctcc 4801 acgttgtggc ggatcttgaa gttcgccatg atgccgttct tctgcttgtc ggcggtgatc 4861 cggacctgcg caaggtcgag gcgctgcaca ctgtcgatgt tcactggagc ataggcggtc 4921 agggcggaaa agtcgtcctg gcctttggcg ctgctggcac tggccaggaa cttcatcgcg 4981 gcggctttgt tcttcgtgcc tttaggcacg acgaggatgt cggccatcac caggttctgc 5041 ttccagctca cgcccactgg cgcgccgtct tcttgcaggg cgtggatgcg accgttccag 5101 aactggccca tgctgacttc gccggaggcc agcagctgct gcgactgtgc accgccgccc 5161 caccagacga tgtctttctt gatggtgtcg agtttcttga aggcgcggtc caggtccagc 5221 gggtagagct tgtcggccgg tacgccgtcg gcgagcaggg ccagttcgag cacgccaggg 5281 ctcggccatt tgtagagggc gcgtttgccg gggtaggtct tggtgtcgaa cagcgcggtc 5341 cagtcctggg gcttgctggc tccgagcttg ccttcgttgt agccgagcac gaaagagaac 5401 aggaacgagc ccacgccatg gtcggagacg aagcgcgggt cgatcttgtc gcgctggatc 5461 accgagaaat ccaggggttc gagcaggcct tcggcggcgg cgcgcaaggc gaagtcggcc 5521 tctacatcca ccacgtccca ctgcacgttg ccgctttcga ccatggcctt gagtttgccg 5581 tagtcggtgg gcccatcctg gaccacggta atgccgctgg ccttgctgaa cgggtcggcc 5641 caggcctgct tctgcgcatc ctgggtgcta ccgccccagc tcacgaaatt gatgctttca 5701 gatctgtcac cagtggaacc tggaacccag agcagcagta cccatagcag gagtgtgtct 5761 gtctccatgg tggcgatatc accggtggta ccactagtat aacttcgtat aggatacttt 5821 atacgaagtt atcattggga ttcttcctat tttgatccaa gcatcaccat cgaccctcta 5881 gtccaaatct caccatcgac ccataacttc gtatagcata cattatacga agttatgtcc 5941 ctcgaagagg ttcgaaacac tgaaaagggc caattctgca aatgatccac tcgagaagct 6001 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta 6061 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc 6121 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga 6181 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac 6241 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc 6301 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc 6361 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg 6421 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc 6481 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc 6541 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg 6601 ataccgtcga cccgggcggc cgctcgagca tgcatctaga gtcgacccgg gcggccgctt 6661 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga 6721 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac c //