LOCUS Exported 6302 bp ds-DNA circular SYN 28-FEB-2024 DEFINITION GABA Sensor. ACCESSION . VERSION . KEYWORDS pAAV.hSynap.iGABASnFR 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) AUTHORS . TITLE Direct Submission JOURNAL Exported Feb 28, 2024 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6302 /organism="synthetic DNA construct" /mol_type="other DNA" 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" primer_bind complement(317..336) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(996..1856) /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 1619..1638 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1857..1961) /gene="bla" /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" primer_bind complement(2124..2145) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2336..2355 /label=F1ori-R /note="F1 origin, reverse primer" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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(5379..5399) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5797..5808) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5917..5933) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5918..5934) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 5963..6084 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5974..5993) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 6028..6047 /label=SV40pA-R /note="SV40 polyA, reverse primer" 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 //