LOCUS Exported 6905 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Medium affinity glutamate sensor. ACCESSION . VERSION . KEYWORDS pAAV.CAG-FLEX.SF-iGluSnFR.A184V 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 26..155 /label=AAV2 ITR promoter 466..743 /label=chicken beta-actin promoter primer_bind 1022..1044 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1106..1125 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind complement(1160..1193) /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(1256..1289) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(1317..1463) /codon_start=1 /product="transmembrane domain from platelet derived growth factor receptor beta" /label=PDGFR-beta TM domain /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ KKPR" CDS complement(1467..1496) /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" primer_bind 1961..1982 /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2085..2106) /label=EGFP-C /note="EGFP, forward primer" sig_peptide complement(3072..3134) /product="leader sequence from mouse immunoglobulin kappa light chain" /label=Ig-kappa leader regulatory 3131..3140 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 3165..3198 /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 3269..3302 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3374..3962 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3427..3447) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3845..3856) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4005..4126 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4016..4035) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4070..4089 /label=SV40pA-R /note="SV40 polyA, reverse primer" repeat_region 4288..4417 /label=AAV2 ITR primer_bind complement(4483..4500) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4654..5242) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4734..4753) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5413..6273) /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 6036..6055 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6274..6378) /gene="bla" /label=AmpR promoter rep_origin complement(6404..6859) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6541..6562) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6753..6772 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 gcgggcctct tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc 61 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag 121 agagggagtg gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg 181 ctacttatct acgtagccat gctctaggaa gagtaccatt gacgtcaata atgacgtatg 241 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 301 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 361 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 421 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca 481 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta 541 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccrggsg 601 gggsggggsg gggsgrgggg sggggsgggg cgaggcggag aggtgcggcg gcagccaatc 661 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata 721 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct 781 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga 841 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg 901 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg 961 ggagcggctc ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg 1021 gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc 1081 ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt 1141 ggcaaagaat tggatcccca taacttcgta taaagtatcc tatacgaagt tatatcaaaa 1201 taggaagacc aatgcttcac catcgacccg aattgccaag catcaccatc gacccataac 1261 ttcgtataat gtatgctata cgaagttata ctagtgctag cggatccgat atcctaacgt 1321 ggcttcttct gccaaagcat gatgaggatg ataagggaga tgatggtgag caccaccagg 1381 gccaggatgg ctgagatcac caccacctta aagggcaagg agtgtggcac cacgatgacc 1441 tcctgcgtgt cctggcccac agcattcaga tcctcttctg agatgagttt ttgttcgtcg 1501 acctgcagtt tcagtgcctt gtcattcggt tctttgaaca gtgctttcat ttcgtctgac 1561 agttcgaaat tcatgttcag ggggttgtta aagttgtact ccagcttgtg ccccaggatg 1621 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc 1681 tcgaacttca cctcggcgcg ggtcttgtag gtgccgtcgt ccttgaagct gatggtgcgc 1741 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg 1801 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc 1861 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgtt ggtggcatcg 1921 ccctcgccct cgccgcgcac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc 1981 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcatgctccc tccggtaccg 2041 cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg 2101 accatgtgat cgcgcttctc gttggggtct ttgctcagca cggactgggt gctcaggtag 2161 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg 2221 gcgagctgca cgctgccgtc ctccacgttg tggcggatct tgaagttcgc cttgatgccg 2281 ttcttctgct tgtcggcggt gatatagacg ttgtggctca ccagaattgg atttttgaac 2341 cacttatcaa accatttttc cgcttcaccg gaggtctgca cctgagcgat ggtgtcatcc 2401 atcagctttt tgaactgcgg atcatcttta cgcaacatac aaccgtaggc ctcctgagac 2461 tgcggcttgc cgacgatttc ccagttgtct ggtttcttcg ctttcgcacg ttcaccggcc 2521 agcagcacgt catccatcat aaaggcaacg gcacgaccgc tttccagggt gcggaaagag 2581 tcaccgtgat ctttggcgct gatgatgcgc atattcattt tttgctcttc attcagtttg 2641 ttgagcaaaa cttcagaggt agtgccggaa gtgacgacta cggctttgtc tttcaggttg 2701 gcaaaatctt tgatatcgcc accctttttg gtcaacaggc gcgtaccgac cacgaaaata 2761 gtgtcagaga aagccgcctg tttttggcgt tcgacgttgt tggtggtaga accacattca 2821 aaatcgaaag tgccgttttg cagcagtgga atacggtttt gtgaggtaat cggaatcagt 2881 tttacctgca agtccggttt gttgagtttc tttttcactg cttcaacaat ggcgttggag 2941 taatcctgcg agtaacccac cactttttgc tgattgtcgt aataagagaa aggcactgaa 3001 gattcacggt gaccgacgac aatcacaccg tttttggcga ttttgtccag cgtgctgcct 3061 gcggcagatc tgtcaccagt ggaacctgga acccagagca gcagtaccca tagcaggagt 3121 gtgtctgtct ccatggtggc gatatcaccg gtggtaccac tagtataact tcgtatagga 3181 tactttatac gaagttatca ttgggattct tcctattttg atccaagcat caccatcgac 3241 cctctagtcc aaatctcacc atcgacccat aacttcgtat agcatacatt atacgaagtt 3301 atgtccctcg aagaggttcg aaacactgaa aagggccaat tctgcaaatg atccactcga 3361 gaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct 3421 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc 3481 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct 3541 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga 3601 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc 3661 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac 3721 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt 3781 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt 3841 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc 3901 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc 3961 gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat acattgatga 4021 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga 4081 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg 4141 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa 4201 cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt 4261 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct 4321 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct 4381 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa tcggccaacg 4441 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 4501 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 4561 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 4621 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 4681 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 4861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 4921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 4981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 5041 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 5101 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 5161 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 5221 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 5281 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 5341 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 5401 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 5461 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 5521 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 5581 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 5641 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 5701 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 5761 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 5821 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 5881 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 5941 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 6001 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 6061 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 6121 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 6181 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 6241 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 6301 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 6361 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat 6421 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 6481 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt 6541 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa 6601 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg 6661 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga 6721 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct 6781 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat 6841 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga 6901 tcggt //