LOCUS Exported 6905 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Tight affinity yellow glutamate sensor. ACCESSION . VERSION . KEYWORDS pAAV.CAG-FLEX.SF-Venus-iGluSnFR.A184S SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6905) AUTHORS Marvin JS, Scholl B, Wilson DE, Podgorski K, Kazemipour A, Muller JA, Schoch S, Quiroz FJU, Rebola N, Bao H, Little JP, Tkachuk AN, Cai E, Hantman AW, Wang SS, DePiero VJ, Borghuis BG, Chapman ER, Dietrich D, DiGregorio DA, Fitzpatrick D, Looger LL TITLE Stability, affinity, and chromatic variants of the glutamate sensor iGluSnFR. JOURNAL Nat Methods. 2018 Nov;15(11):936-939. doi: 10.1038/s41592-018-0171-3. Epub 2018 Oct 30. PUBMED 30377363 REFERENCE 2 (bases 1 to 6905) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6905 /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 2367..2388 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2491..2512) /label=EGFP-N /note="EGFP, reverse primer" CDS 2977..3006 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 3010..3156 /codon_start=1 /product="transmembrane domain from platelet derived growth factor receptor beta" /label=PDGFR-beta TM domain /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR" protein_bind 3184..3217 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 3280..3313 /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(3348..3367) /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" primer_bind complement(3429..3451) /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" promoter complement(3730..4007) /label=chicken beta-actin promoter repeat_region 4318..4447 /label=AAV2 ITR rep_origin 4519..4974 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4606..4625) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4816..4837 /label=F1ori-F /note="F1 origin, forward primer" promoter 5000..5104 /gene="bla" /label=AmpR promoter CDS 5105..5965 /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(5323..5342) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6136..6724 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6625..6644 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6878..6895 /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 cagatctgcc gcaggcagca cgctggacaa aatcgccaaa 1441 aacggtgtga ttgtcgtcgg tcaccgtgaa tcttcagtgc ctttctctta ttacgacaat 1501 cagcaaaaag tggtgggtta ctcgcaggat tactccaacg ccattgttga agcagtgaaa 1561 aagaaactca acaaaccgga cttgcaggta aaactgattc cgattacctc acaaaaccgt 1621 attccactgc tgcaaaacgg cactttcgat tttgaatgtg gttctaccac caacaacgtc 1681 gaacgccaaa aacaggcggc tttctctgac actattttcg tggtcggtac gcgcctgttg 1741 accaaaaagg gtggcgatat caaagatttt gccaacctga aagacaaagc cgtagtcgtc 1801 acttccggca ctacctctga agttttgctc aacaaactga atgaagagca aaaaatgaat 1861 atgcgcatca tcagcgccaa agatcacggt gactctttcc gcaccctgga aagcggtcgt 1921 gccgttgcct ttatgatgga tgacagcctg ctggccggtg aacgtgcgaa agcgaagaaa 1981 ccagacaact gggaaatcgt cggcaagccg cagtctcagg aggcctacgg ttgtatgttg 2041 cgtaaagatg atccgcagtt caaaaagctg atggatgaca ccatcgctca ggtgcagacc 2101 tccggtgaag cggaaaaatg gtttgataag tggttcaaaa atccaattct ggtgagccac 2161 aacgtctata tcaccgccga caagcagaag aacggcatca aggcgaactt caagatccgc 2221 cacaacgtgg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc 2281 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctatcagtc cgtgctgagc 2341 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 2401 atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat gagcaagggc 2461 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 2521 cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg 2581 aagctcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 2641 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc 2701 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcagcttcaa ggacgacggc 2761 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 2821 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 2881 tttaacaacc ccctgaacat gaatttcgaa ctgtcagacg aaatgaaagc actgttcaaa 2941 gaaccgaatg acaaggcact gaaactgcag gtcgacgaac aaaaactcat ctcagaagag 3001 gatctgaatg ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc 3061 tttaaggtgg tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt 3121 atcatcctca tcatgctttg gcagaagaag ccacgttagg atatcggatc cgctagcact 3181 agtataactt cgtatagcat acattatacg aagttatggg tcgatggtga tgcttggcaa 3241 ttcgggtcga tggtgaagca ttggtcttcc tattttgata taacttcgta taggatactt 3301 tatacgaagt tatggggatc caattctttg ccaaaatgat gagacagcac aataaccagc 3361 acgttgccca ggagctgtag gaaaaagaag aaggcatgaa catggttagc agaggctcta 3421 gagccgccgg tcacacgcca gaagccgaac cccgccctgc cccgtccccc ccgaaggcag 3481 ccgtcccccc gcggacagcc ccgagccgct ccccccgcac aaagggccct cccggagccc 3541 ctcaaggctt tcacgcagcc acagaaaaga aacaagccgt cattaaacca agcgctaatt 3601 acagcccgga ggagaagggc cgtcccgccc gctcacctgt gggagtaacg cggtcagtca 3661 gagccggggc gggcggcgcg aggcggcggc ggagcggggc acggggcgaa ggcagcgcgc 3721 agcgactccc gcccgccgcg cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc 3781 cataaaagga aactttcgga gcgcgccgct ctgattggct gccgccgcac ctctccgcct 3841 cgccscgccc csccccycsc cccgccccgc cccgcctggc gcgcgccccc cccccccccc 3901 cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg ggggtgggga 3961 ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca tggtaatagc 4021 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc 4081 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg 4141 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca 4201 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatggtac 4261 tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt aactacaagg 4321 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg 4381 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag 4441 cgcgcagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca 4501 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 4561 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 4621 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 4681 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 4741 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 4801 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 4861 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 4921 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg 4981 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 5041 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 5101 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct 5161 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg 5221 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg 5281 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt 5341 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga 5401 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga 5461 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac 5521 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg 5581 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac 5641 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct 5701 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct 5761 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg 5821 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat 5881 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg 5941 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat 6001 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 6061 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 6121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 6181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 6241 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta 6301 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 6361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 6421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 6481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 6541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 6601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 6661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 6721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca 6781 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 6841 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc 6901 ggaag //