LOCUS Exported 5368 bp ds-DNA circular SYN 17-JUN-2022 DEFINITION Red fluorescent calcium sensor protein. ACCESSION . VERSION . KEYWORDS pGP-CMV-NES-jRGECO1a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5368) AUTHORS Dana H, Mohar B, Sun Y, Narayan S, Gordus A, Hasseman JP, Tsegaye G, Holt GT, Hu A, Walpita D, Patel R, Macklin JJ, Bargmann CI, Ahrens MB, Schreiter ER, Jayaraman V, Looger LL, Svoboda K, Kim DS TITLE Sensitive red protein calcium indicators for imaging neural activity. JOURNAL Elife. 2016 Mar 24;5. pii: e12727. doi: 10.7554/eLife.12727. PUBMED 27011354 REFERENCE 2 (bases 1 to 5368) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 17, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5368 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 15..34 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 137..931 /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(191..210) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 801..820 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(1119..1138) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 1163..1210 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 1539..2127 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2028..2047 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 2302..2605 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2606..2809 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2760..2780 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 2929..2946 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 2950..2982 /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 2986..3009 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 3037..3093 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKAGHAVRAIGRLSS" primer_bind 3789..3808 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 4397..4518 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4434..4453) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4488..4507 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4525..4980) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4662..4683) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4874..4893 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5007..5111 /gene="bla" /label=AmpR promoter promoter 5113..102 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(5136..5156) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 5321..88 /label=SV40 ori /note="SV40 origin of replication" ORIGIN 1 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 61 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg 121 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 181 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 241 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 301 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt 361 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 421 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 481 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 541 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 601 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 661 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 721 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 781 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 841 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 901 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac 961 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 1021 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 1081 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata 1141 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg 1201 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg 1261 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt 1321 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca 1381 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 1441 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 1501 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 1561 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 1621 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 1681 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 1741 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 1801 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 1861 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 1921 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 1981 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 2041 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 2101 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 2161 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 2221 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca 2281 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc 2341 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat 2401 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt 2461 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc 2521 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta 2581 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg 2641 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt 2701 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac 2761 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa 2821 ccgtcagatc cgctagcgct accggactca gatctcgcca ccatgctgca gaacgagctt 2881 gctcttaagt tggctggact tgatattaac aagactggag gaggttctca tcatcatcat 2941 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat 3001 gacgataagg atctcgcaac aatggtcgac tcatcacgtc gtaagtggaa taaggcaggt 3061 cacgcagtca gagctatagg tcggctgagc tcacccgtgg tttccgagcg gatgtacccc 3121 gaggacggcg ccctgaagag cgagatcaag aaggggctga ggctgaagga cggcggccac 3181 tacgccgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc 3241 tacatcgtgg acatcaagtt ggacatcgtg tcccacaacg aggactacac catcgtggaa 3301 cagtgcgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaaggga 3361 ggtacaggcg ggagtctggt gagcaagggc gaggaggata acatggccat catcaaggag 3421 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag 3481 ggcgagggcg agggccgccc ctacgaggcc tttcagaccg ctaagctgaa ggtgaccaag 3541 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag 3601 gcctacatta agcacccagc cgacatcccc gactacttca agctgtcctt ccccgagggc 3661 ttcaggtggg agcgcgtgat gaacttcgag gacggcggca ttattcacgt gaaccaggac 3721 tcctccctgc aggacggcgt attcatctac aaggtgaagc tgcgcggcac caacttcccc 3781 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggctacgcg tgacgacctg 3841 actgaagagc agatcgcaga atttaaagag gctttctccc tatttgacaa ggacggggat 3901 gggacgataa caaccaagga gcttgggacg gtgttccgat cgctggggca gaaccccaca 3961 gaagcagagc tgcaggacat gatcaatgaa gtagatgccg acggtgacgg cacattcgac 4021 ttccctgagt tcctgacgat gatggcaaga aaaatgaatg acacagacag tgaagaggaa 4081 attcgcgaag cgttccgcgt gtttgataag gacggcaatg gctacatcgg cgcagcagag 4141 cttcgccacg tgatgacaga ccttggagag aagttaacag atgaggaggt tgatgaaatg 4201 atcagggtag cagacatcga tggggatggt caggtaaact acgaagagtt tgtacaaatg 4261 atgacagcga agtaggcggc cgcgactcta gatcataatc agccatacca catttgtaga 4321 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa 4381 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag 4441 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa 4501 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt 4561 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt 4621 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc 4681 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg 4741 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac 4801 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg 4861 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag 4921 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt 4981 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 5041 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 5101 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg 5161 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 5221 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 5281 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc 5341 cagttccgcc cattctccgc cccatggc //