LOCUS Exported 7105 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION GABA Sensor. ACCESSION . VERSION . KEYWORDS pAAV.hSyn-FLEX.iGABASnFR.F102G SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7105) 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 7105) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7105 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(12..29) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(183..771) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(263..282) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(942..1802) /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 1565..1584 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1803..1907) /gene="bla" /label=AmpR promoter rep_origin complement(1933..2388) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2070..2091) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2282..2301 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 2570..2699 /label=AAV2 ITR polyA_signal 2861..2982 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2898..2917) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2952..2971 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" misc_feature 3060..3648 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS 3166..3177 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind 3575..3595 /label=WPRE-R /note="WPRE, reverse primer" protein_bind complement(3720..3753) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(3824..3857) /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 3882..3891 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" sig_peptide 3888..3950 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader primer_bind 4994..5015 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(5118..5139) /label=EGFP-N /note="EGFP, reverse primer" CDS 5646..5675 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 5679..5825 /codon_start=1 /product="transmembrane domain from platelet derived growth factor receptor beta" /label=PDGFR-beta TM domain /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR" protein_bind 5853..5886 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 5949..5982 /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(5993..6012) /label=pMT2-F /note="Synthetic intron, forward primer" intron 5995..6127 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" repeat_region 6720..6849 /label=AAV2 ITR primer_bind complement(6887..6903) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6887..6903) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6900..6922) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6911..6927 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6935..6965) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6980..7001 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 61 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 121 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 181 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 241 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 301 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 361 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 421 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 481 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 541 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 601 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 661 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 721 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 781 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 841 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 901 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 961 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 1021 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 1081 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 1141 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 1201 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 1261 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 1321 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 1381 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 1441 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 1501 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 1561 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 1621 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 1681 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 1741 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 1801 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 1861 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 1921 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt 1981 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag 2041 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga 2101 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg 2161 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc 2221 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg 2281 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc 2341 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat 2401 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc 2461 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cggcgccagg gcggcctcng 2521 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tgcgcgctcg 2581 ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc 2641 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt 2701 gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta ggaagatcct 2761 tatcgatttt accacatttg tagaggtttt acttgcttta aaaaacctcc cacatctccc 2821 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta 2881 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 2941 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gctcgaagcg 3001 gccgcccggg tcgactctag atgcatgctc gagcggccgc ccgggtcgac ggtatcgatg 3061 cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa gacgcggaag 3121 aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga gggccgaagg 3181 gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga gcagccaagg 3241 aaaggacgat gatttccccg acaacaccac ggaattgtca gtgcccaaca gccgagcccc 3301 tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag ggagggggaa 3361 agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag tgggggttgc 3421 gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca actcctcata 3481 aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac gggaagcaat 3541 agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag caacatagtt 3601 aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattat cgataagctt 3661 ctcgagtgga tcatttgcag aattggccct tttcagtgtt tcgaacctct tcgagggaca 3721 taacttcgta taatgtatgc tatacgaagt tatgggtcga tggtgagatt tggactagag 3781 ggtcgatggt gatgcttgga tcaaaatagg aagaatccca atgataactt cgtataaagt 3841 atcctatacg aagttatact agtggtacca ccggtgatat cgccaccatg gagacagaca 3901 cactcctgct atgggtactg ctgctctggg ttccaggttc cactggtgac agatctgaaa 3961 gcatcaattt cgtgagctgg ggcggtagca cccaggatgc gcagaagcag gcctgggccg 4021 acccgttcag caaggccagc ggcattaccg tggtccagga tgggcccacc gactacggca 4081 aactcaaggc catggtcgaa agcggcaacg tgcagtggga cgtggtggat gtagaggccg 4141 acttcgcctt gcgcgccgcc gccgaaggcc tgctcgaacc cctggatttc tcggtgatcc 4201 agcgcgacaa gatcgacccg cgcttcgtct ccgaccatgg cgtgggctcg ttcctgggtt 4261 ctttcgtgct cggctacaac gaaggcaagc tcggagccag caagccccag gactggaccg 4321 cgctgttcga caccaagacc taccccggca aacgcgccct ctacaaatgg ccgagccctg 4381 gcgtgctcga actggccctg ctcgccgacg gcgtaccggc cgacaagctc tacccgctgg 4441 acctggaccg cgccttcaag aaactcgaca ccatcaagaa agacatcgtc tggtggggcg 4501 gcggtgcaca gtcgcagcag ctgctggcct ccggcgaagt cagcatgggc cagttctgga 4561 acggtcgcat ccacgccctg caagaagacg gcgcgccagt gggcgtgagc tggaagcaga 4621 acctggtgat ggccgacatc ctcgtcgtgc ctaaaggcac gaagaacaaa gccgccgcga 4681 tgaagttcct ggccagtgcc agcagcgcca aaggccagga cgacttttcc gccctgaccg 4741 cctatgctcc agtgaacatc gacagtgtgc agcgcctcga ccttgcgcag gtccggatca 4801 ccgccgacaa gcagaagaac ggcatcatgg cgaacttcaa gatccgccac aacgtggagg 4861 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 4921 tgctgctgcc cgacaaccac tacctgagca cccagtccgt gctgagcaaa gaccccaacg 4981 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca 5041 tggacgagct gtacaagggc ggtaccggag ggagcatgag caagggcgag gagctgttca 5101 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg 5161 tgcgcggcga gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca 5221 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc 5281 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc 5341 ccgaaggcta cgtccaggag cgcaccatca gcttcaagga cgacggcacc tacaagaccc 5401 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg 5461 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactgg aacgccaacc 5521 tggcaccgaa cctgccgacc gcctacgtca aggatcagat caccctcgat ttcgcctact 5581 gggccaagaa cggtccggcc attgcgacac ggtggaatga atggctagtc aaactgcagg 5641 tcgacgaaca aaaactcatc tcagaagagg atctgaatgc tgtgggccag gacacgcagg 5701 aggtcatcgt ggtgccacac tccttgccct ttaaggtggt ggtgatctca gccatcctgg 5761 ccctggtggt gctcaccatc atctccctta tcatcctcat catgctttgg cagaagaagc 5821 cacgttagga tatcggatcc gctagcacta gtataacttc gtatagcata cattatacga 5881 agttatgtgt cgatggtgat gcttggcaat tcgggtcgat ggtgaagcat tggtcttcct 5941 attttgatat aacttcgtat aggatacttt atacgaagtt atggggatca tcacctgtgg 6001 agagaaaggc aaagtggatg tcagtaagac caataggtgc ctatcagaaa cgcaagagtc 6061 ttctctgtct cgacaagccc agtttctatt ggtctcctta aacctgtctt gtaaccttga 6121 tacttacctg atccaacggc tgcgacttgg ggcagggggt cctaggggtg gtctggccag 6181 gagccgcggg ggcggactgc gcggtgccca ggagcacagc tgcgctctca ggcacgacac 6241 gactcctccg ctgcccaccg cagactgagg cagcgctgag tcgccggcgc cgcagcgcag 6301 atggtcgcgc ccgtgccccc ctatctcgcg cctcgcgtgg tgcggtccgg ctgggccggc 6361 ggcggcgcgg acgcgaccaa ggtggccggg aaggggagtt tgcgggggac cggcgagtga 6421 cgtcagcgcg ccttcagtgc tgaggcggcg gtggcgcgcg ccgccaggcg ggggcgaagg 6481 cactgtccgc ggtgctgaag ctggcagtgc gcacgcgcct cgccgcatcc tgtttcccct 6541 ccccctctct gataggggat gcgcaatttg gggaatgggg gttgggtgct tgtccagtgg 6601 gtcggggtcg gtcgtcaatt taaatactga aaagggcgaa ttccagcaca ctggagctcg 6661 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa 6721 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc 6781 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg 6841 agcgcgcagc tggcgtaata gcgaagtgga gcctggcgta atcatggtca tagctgtttc 6901 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt 6961 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc 7021 ccgctttcca gtcgggaaac ctgtcgcgca gctgcattaa tgaatcggcc aacgcgcggg 7081 gagaggcggt ttgcgtattg ggcgc //