LOCUS Exported 5341 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Red fluorescent calcium sensor protein. ACCESSION . VERSION . KEYWORDS pGP-CMV-NES-jRCaMP1a SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5341) 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 5341) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5341 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 688..705 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 709..741 /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 745..768 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" CDS 796..852 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" polyA_signal 2129..2250 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2166..2185) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2220..2239 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2257..2712) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2394..2415) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2606..2625 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2739..2843 /gene="bla" /label=AmpR promoter promoter 2845..3202 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2868..2888) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3053..3188 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3115..3134 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3237..4031 /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(3291..3310) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3901..3920 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4219..4238) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4263..4310 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4639..5227 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5128..5147 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgccac catgctgcag aacgagcttg ctcttaagtt ggctggactt 661 gatattaaca agactggagg aggttctcat catcatcatc atcatggtat ggctagcatg 721 actggtggac agcaaatggg tcgggatctg tacgacgatg acgataagga tctcgcaaca 781 atggtcgact catcacgtcg taagtggaat aagacaggtc acgcagtcag agctataggt 841 cggctgagct cagcgatcaa ctgcgaaatg atgtacccag cggatggtgg tctgcgtggt 901 tacactcaca tggcgctgaa agttgatggc ggcggtcacc tgtcctgttc tttcgtgacc 961 acctaccgct ccaaaaagac tgtcggcaac attaagatgc ctgccattca tagcgtcagc 1021 caccgtctgg agcgcctgga ggagagcgat aacgaaatgt ttgtcgtaca gcgtgaacac 1081 gcagttgcca agtttgtggg cctgggtggt ggcggcggta ccggagggag catgaactcc 1141 ctgatcaagg agaacatgcg tatgaaagtg gttctggaag gctccgtaaa cggccaccag 1201 ttcaaatgca ctggtgaagg cgaaggcaac ccgtatatgg gcacccagac tatgcgtatc 1261 aaagtgatcg agggtggtcc gctgccgttt gcgttcgaca tcctggcgac gtcctttatg 1321 tatggctccc gtaccttcat caaatatccg aaaggcatcc cggatttctt taagcagtcc 1381 ttcccggaag gttttacctg ggaacgtgtg acccgttacg aagacggcgg cgtaattacc 1441 gttatgcaag acacgtctct ggaggatggc tgcctggtgt atcacgtgca ggttcgcggt 1501 gtgaacttcc cgagcaatgg tgctgtaatg caaaagaaaa ccaaaggttg ggagcctacg 1561 gactcccaaa ccactgaaga gcagatcgca gaatttaaag aggctttctc cctatttgac 1621 aaggacgggg atgggacaat aacaaccaag gagctgggga cggtgatgcg gtctctgggg 1681 cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac 1741 ggcacaatcg acttccctga gttcctgatt atgatggcaa gaaaaatgaa atacacagac 1801 agtgaagaag aaattagaga agcgttcggc gtgtttgata aggatggcaa tggctacatc 1861 agtgcagcag agcttcgcca cgtgatgaca aaccttggag agaagttaac agatgaagag 1921 gttgatgaaa tgatcaggga agcagacagc gatggggatg gtcaggtaaa ctacgaagag 1981 tttgtacaaa tgatgacagc gaagtaggcg gccgcgactc tagatcataa tcagccatac 2041 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa 2101 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa 2161 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 2221 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt 2281 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg 2341 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt 2401 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct 2461 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt 2521 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa 2581 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc 2641 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc 2701 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat 2761 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc 2821 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca 2881 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct 2941 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 3001 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc 3061 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta 3121 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt 3181 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga 3241 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 3301 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 3361 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag 3421 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 3481 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 3541 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 3601 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 3661 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 3721 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg 3781 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 3841 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 3901 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 3961 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 4021 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 4081 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 4141 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 4201 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat 4261 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg 4321 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata 4381 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct 4441 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta 4501 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa 4561 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 4621 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 4681 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 4741 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc 4801 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 4861 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 4921 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 4981 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 5041 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 5101 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 5161 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 5221 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 5281 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca 5341 t //