LOCUS Exported 5308 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Mammalian expression of ultrasensitive protein calcium sensor. ACCESSION . VERSION . KEYWORDS pGP-CMV-jGCaMP7c variant 1513 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5308) AUTHORS Dana H, Sun Y, Mohar B, Hulse BK, Kerlin AM, Hasseman JP, Tsegaye G, Tsang A, Wong A, Patel R, Macklin JJ, Chen Y, Konnerth A, Jayaraman V, Looger LL, Schreiter ER, Svoboda K, Kim DS TITLE High-performance calcium sensors for imaging activity in neuronal populations and microcompartments. JOURNAL Nat Methods. 2019 Jul;16(7):649-657. doi: 10.1038/s41592-019-0435-6. Epub 2019 Jun 17. PUBMED 31209382 REFERENCE 2 (bases 1 to 5308) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5308 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 124..427 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 428..631 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 582..602 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 679..688 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 694..711 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 715..747 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 751..774 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" regulatory 781..790 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 802..858 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" primer_bind 1071..1092 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(1198..1219) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1459..1478) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" polyA_signal 2159..2280 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2196..2215) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2250..2269 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2287..2742) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2424..2445) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2636..2655 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2769..2873 /gene="bla" /label=AmpR promoter promoter 2875..3232 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2898..2918) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3083..3218 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3145..3164 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3267..4061 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3321..3340) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3931..3950 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4249..4268) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4293..4340 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4669..5257 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5158..5177 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg 61 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt 121 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc 181 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg 241 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat 301 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca 361 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat 421 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg 481 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca 541 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg 601 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg 661 ctaccggact cagatctcgc caccatgggt tctcatcatc atcatcatca tggtatggct 721 agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga taaggatctc 781 gccaccatgg tcgactcatc acgtcgtaag tggaataaga caggtcacgc agtcagagct 841 ataggtcggc tgagctcaca gcctaacgtc tatatcaagg ccgacaagca gaagaacggc 901 atcaaggcga acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgcctac 961 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 1021 ctgagcgtgc agtccaaact ttcgaaagac cccaacgaga agcgcgatca catggtcctg 1081 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt 1141 accggaggga gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 1201 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc 1261 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 1321 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc 1381 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag 1441 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 1501 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1561 atcctggggc acaagctgga gtacaacctt cctgaccaac tgactgaaga gcagatcgca 1621 gaatttaaag aggctttctc cctatttgac aaggacgggg atgggacaat aacaaccaag 1681 gagctgggga cggtgatgcg ttctctgggg cagaacccca cagaagcaga gctgcaggac 1741 atgatcaatg aagtagatgc cgacggtgac ggcacaatcg acttccctga gttcctgaca 1801 atgatggcaa gaaaagggag ctacagggac agtgaagaag aaattagaga agcgttcggt 1861 gtgtttgata aggatggcaa tggctacatc agtgcagcag agcttcgcca cgtgatgaac 1921 aaccttggag agaagttaac agatgaagag gttgatgaaa tgatcaggga agcagacatc 1981 gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc gaagtgagcg 2041 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa 2101 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa 2161 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 2221 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 2281 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 2341 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 2401 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 2461 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat 2521 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 2581 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 2641 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 2701 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg 2761 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 2821 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg 2881 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc 2941 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc 3001 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat 3061 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc 3121 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga 3181 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc 3241 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc 3301 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 3361 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 3421 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca 3481 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 3541 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 3601 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 3661 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 3721 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 3781 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 3841 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 3901 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 3961 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 4021 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga 4081 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt 4141 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg 4201 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa 4261 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca 4321 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga 4381 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca 4441 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc 4501 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa 4561 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 4621 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 4681 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 4741 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 4801 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 4861 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 4921 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 4981 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 5041 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 5101 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 5161 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 5221 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 5281 acggttcctg gccttttgct ggcctttt //