LOCUS Exported 7643 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Medium affinity glutamate sensor. ACCESSION . VERSION . KEYWORDS pAAV.CAG-FLEX.SF-iGluSnFR.A184V.mRuby3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7643) 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 7643) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7643 /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 3919..3952 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 4015..4048 /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(4083..4102) /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" primer_bind complement(4164..4186) /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" promoter complement(4466..4743) /label=chicken beta-actin promoter repeat_region 5056..5185 /label=AAV2 ITR rep_origin 5257..5712 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5344..5363) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5554..5575 /label=F1ori-F /note="F1 origin, forward primer" promoter 5738..5842 /gene="bla" /label=AmpR promoter CDS 5843..6703 /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(6061..6080) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6874..7462 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7363..7382 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7616..7633 /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 tgacgtgctg 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 gcacccagtc 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 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 2641 acctacggcg tgcagtgctt cagccgctac 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 ccacgttcta gaggctccgg aaatgctgtg 3181 ggccaggaca cgcaggagcg tatggtgtct aagggcgaag agctgatcaa ggaaaatatg 3241 cgtatgaagg tggtcatgga aggttcggtc aacggccacc aattcaaatg cacaggtgaa 3301 ggagaaggca ggccgtacga gggagtgcaa accatgagga tcaaagtcat cgagggagga 3361 cccctgccat ttgcctttga cattcttgcc acgtcgttca tgtatggcag ccgtactttt 3421 atcaagtacc cggcagacat tcctgatttc tttaaacagt cctttcctga gggttttact 3481 tgggaaagag ttacgagata cgaagatggt ggagtcgtca ccgtcacgca ggacaccagc 3541 cttgaggatg gcgaactcgt ttacaacgtc aaagtcagag gggtaaactt tccctccaat 3601 ggtcccgtga tgcagaagaa gaccaagggt tgggagccta atacagagat gatgtatcca 3661 gcagatggtg gtctgagggg atacactgat attgcactga aagttgatgg tggtggccat 3721 ctgcattgca atttcgtaac aacttacagg tcaaaaaaga ccgtcgggaa catcaagatg 3781 cccggtgtcc atgccgttga tcaccgcctg gaaaggatag aggaaagtga caatgaaacg 3841 tacgtagtac aacgcgaagt cgcagttgcc aagtactcca atcttggtgg ttaggatatc 3901 ggatccgcta gcactagtat aacttcgtat agcatacatt atacgaagtt atgggtcgat 3961 ggtgatgctt ggcaattcgg gtcgatggtg aagcattggt cttcctattt tgatataact 4021 tcgtatagga tactttatac gaagttatgg ggatccaatt ctttgccaaa atgatgagac 4081 agcacaataa ccagcacgtt gcccaggagc tgtaggaaaa agaagaaggc atgaacatgg 4141 ttagcagagg ctctagagcc gccggtcaca cgccagaagc cgaaccccgc cctgccccgt 4201 cccccccgaa ggcagccgtc cccccgcgga cagccccgag ccgctccccc cgcacaaagg 4261 gccctcccgg agcccctcaa ggctttcacg cagccacaga aaagaaacaa gccgtcatta 4321 aaccaagcgc taattacagc ccggaggaga agggccgtcc cgcccgctca cctgtgggag 4381 taacgcggtc agtcagagcc ggggcgggcg gcgcgaggcg gcggcggagc ggggcacggg 4441 gcgaaggcag cgtcgcagcg actcccgccc gccgcgcgct tcgcttttta tagggccgcc 4501 gccgccgccg cctcgccata aaaggaaact ttcggagcgc gccgctctga ttggctgccg 4561 ccgcacctct ccgcctcgcc ccgccccgcc cctcgccccg ccccgccccg cctggcgcgc 4621 gccccccccc cccccccgcc cccatcgctg cacaaaataa ttaaaaaata aataaataca 4681 aaattggggg tggggagggg ggggagatgg ggagagtgaa gcagaacgtg gggctcacct 4741 cgaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata 4801 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg 4861 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact 4921 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta 4981 ttgacgtcaa tggtacgatc ttcctagagc atggctacgt agataagtag catggcgggt 5041 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc 5101 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg 5161 cctcagtgag cgagcgagcg cgcagctggc gtaatagcga agaggcccgc accgatcgcc 5221 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa 5281 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 5341 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 5401 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 5461 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 5521 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 5581 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 5641 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 5701 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt 5761 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca 5821 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt 5881 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga 5941 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa 6001 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct 6061 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat 6121 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga 6181 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc 6241 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat 6301 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa 6361 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac 6421 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa 6481 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc 6541 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc 6601 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag 6661 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta 6721 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa 6781 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 6841 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 6901 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 6961 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 7021 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 7081 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 7141 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 7201 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 7261 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 7321 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 7381 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 7441 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 7501 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 7561 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 7621 gtcagtgagc gaggaagcgg aag //