LOCUS Exported 9600 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Myofliament localised red genetically encoded calcium sensor in mammalian expression vector used for AdEasy adenoviral DNA recombination.. ACCESSION . VERSION . KEYWORDS pShuttle CMV RGECO-TnT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9600) AUTHORS Sparrow AJ, Sievert K, Patel S, Chang YF, Broyles CN, Brook FA, Watkins H, Geeves MA, Redwood CS, Robinson P, Daniels MJ TITLE Measurement of Myofilament-Localised Calcium Dynamics in Adult Cardiomyocytes and the Effect of Hypertrophic Cardiomyopathy Mutations. JOURNAL Circ Res. 2019 Feb 8. doi: 10.1161/CIRCRESAHA.118.314600. PUBMED 30732532 REFERENCE 2 (bases 1 to 9600) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9600 /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 642..698 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKAGHAVRAIGRLSS" primer_bind 1394..1413 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2909..3030 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2946..2965) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3000..3019 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" repeat_region 6221..6323 /label=ITR /note="inverted terminal repeat of human adenovirus serotype 5" primer_bind complement(6412..6429) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6583..7171) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6663..6682) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 8003..8797 /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(8057..8076) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 8667..8686 /label=Neo-F /note="Neomycin resistance gene, forward primer" rep_origin complement(8845..9225) /direction=LEFT /label=M13 ori /note="M13 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8907..8928) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 9119..9138 /label=F1ori-R /note="F1 origin, reverse primer" repeat_region 9263..9365 /label=ITR /note="inverted terminal repeat of human adenovirus serotype 5" misc_signal 9453..3 /label=Ad5 Psi /note="packaging signal for adenovirus serotype 5" ORIGIN 1 aatactgtaa 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 gctagagatc 601 tggtaccgtc gacgcggccg cccaccatgg tagactcatc acgtcgtaag tggaataagg 661 caggtcacgc agtcagagct ataggtcggc tgagctcacc cgtggtttcc gagcggatgt 721 accccgagga cggcgccctc aagagcgaga tcaagaaggg gctgaggctg aaggacggcg 781 gccactacgc cgccgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg 841 gcgcctacat cgtagacatc aagttggaca tcgtgtccca caacgaggac tacaccatcg 901 tggaacagtg cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctataca 961 agggaggtac aggcgggagt ctggtgagca agggcgagga ggataacatg gccatcatca 1021 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga 1081 tcgagggcga gggcgagggc cgcccctacg aggcctttca gaccgctaag ctgaaggtga 1141 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct 1201 ccaaggccta cattaagcac ccagccgaca tccccgacta cttcaagctg tccttccccg 1261 agggcttcag gtgggagcgc gtgatgaact tcgaggacgg cggcattatt cacgttaacc 1321 aggactcctc cctgcaggac ggcgtattca tctacaaggt gaagctgcgc ggcaccaact 1381 tcccccccga cggccccgta atgcagaaga agaccatggg ctgggaggct acgcgtgacc 1441 aactgactga ggagcagatc gcagaattta aagaggcttt ctccctattt gacaaggacg 1501 gggatgggac gataacaacc aaggagctgg ggacggtgat gcggtctctg gggcagaacc 1561 ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt gacggcacat 1621 tcgacttccc tgagttcctg acgatgatgg caagaaaaat gaatgacaca gacagtgaag 1681 aggaaattag agaagcgttc cgcgtgtttg ataaggacgg caatggctac atcggcgcag 1741 cagagcttcg ccacgtgatg acagaccttg gagagaagtt aacagatgag gaggttgatg 1801 aaatgatcag ggtagcagac atcgatgggg atggtcaggt aaactacgaa gagtttgtcc 1861 aaatgatgac agcgaaggga tccatgtctg acatagaaga ggtggtggaa gagtacgagg 1921 aggaggagca ggaagaagca gctgttgaag agcaggagga ggcagcggaa gaggatgctg 1981 aagcagaggc tgagaccgag gagaccaggg cagaagaaga tgaagaggaa gaggaagcaa 2041 aggaggctga agatggccca atggaggagt ccaaaccaaa gcccaggtcg ttcatgccca 2101 acttggtgcc tcccaagatc cccgatggag agagagtgga ctttgatgac atccaccgga 2161 agcgcatgga gaaggacctg aatgagttgc aggcgctgat cgaggctcac tttgagaaca 2221 ggaagaaaga ggaggaggag ctcgtttctc tcaaagacag gatcgagaga cgtcgggcag 2281 agcgggccga gcagcagcgc atccggaatg agcgggagaa ggagcggcag aaccgcctgg 2341 ctgaagagag ggctcgacga gaggaggagg agaaccgtcg taaggctgag gatgaggccc 2401 ggaagaagaa ggctttgtcc aacatgatgc attttggggg ttacatccag aagcaggccc 2461 agacagagcg gaaaagtggg aagaggcaga ctgagcggga aaagaagaag aagattctgg 2521 ctgagcgtcg taaggtgctg gccattgacc acctgaatga agatcagctg agggagaagg 2581 ccaaggagct gtggcagagc atctataact tggaggcaga gaagttcgac ctgcaggaga 2641 agttcaagca gcagaaatat gagatcaatg ttctccgaaa caggatcaac gataaccaga 2701 aagtctccaa gacccgcggg aaggctaaag tcaccgggcg ctggaaatag ctcgagccta 2761 agcttctaga taagatatcc gatccaccgg atctagataa ctgatcataa tcagccatac 2821 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa 2881 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa 2941 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 3001 tggtttgtcc aaactcatca atgtatctta acgcggatct gggcgtggtt aagggtggga 3061 aagaatatat aaggtggggg tcttatgtag ttttgtatct gttttgcagc agccgccgcc 3121 gccatgagca ccaactcgtt tgatggaagc attgtgagct catatttgac aacgcgcatg 3181 cccccatggg ccggggtgcg tcagaatgtg atgggctcca gcattgatgg tcgccccgtc 3241 ctgcccgcaa actctactac cttgacctac gagaccgtgt ctggaacgcc gttggagact 3301 gcagcctccg ccgccgcttc agccgctgca gccaccgccc gcgggattgt gactgacttt 3361 gctttcctga gcccgcttgc aagcagtgca gcttcccgtt catccgcccg cgatgacaag 3421 ttgacggctc ttttggcaca attggattct ttgacccggg aacttaatgt cgtttctcag 3481 cagctgttgg atctgcgcca gcaggtttct gccctgaagg cttcctcccc tcccaatgcg 3541 gtttaaaaca taaataaaaa accagactct gtttggattt ggatcaagca agtgtcttgc 3601 tgtctttatt taggggtttt gcgcgcgcgg taggcccggg accagcggtc tcggtcgttg 3661 agggtcctgt gtattttttc caggacgtgg taaaggtgac tctggatgtt cagatacatg 3721 ggcataagcc cgtctctggg gtggaggtag caccactgca gagcttcatg ctgcggggtg 3781 gtgttgtaga tgatccagtc gtagcaggag cgctgggcgt ggtgcctaaa aatgtctttc 3841 agtagcaagc tgattgccag gggcaggccc ttggtgtaag tgtttacaaa gcggttaagc 3901 tgggatgggt gcatacgtgg ggatatgaga tgcatcttgg actgtatttt taggttggct 3961 atgttcccag ccatatccct ccggggattc atgttgtgca gaaccaccag cacagtgtat 4021 ccggtgcact tgggaaattt gtcatgtagc ttagaaggaa atgcgtggaa gaacttggag 4081 acgcccttgt gacctccaag attttccatg cattcgtcca taatgatggc aatgggccca 4141 cgggcggcgg cctgggcgaa gatatttctg ggatcactaa cgtcatagtt gtgttccagg 4201 atgagatcgt cataggccat ttttacaaag cgcgggcgga gggtgccaga ctgcggtata 4261 atggttccat ccggcccagg ggcgtagtta ccctcacaga tttgcatttc ccacgctttg 4321 agttcagatg gggggatcat gtctacctgc ggggcgatga agaaaacggt ttccggggta 4381 ggggagatca gctgggaaga aagcaggttc ctgagcagct gcgacttacc gcagccggtg 4441 ggcccgtaaa tcacacctat taccggctgc aactggtagt taagagagct gcagctgccg 4501 tcatccctga gcaggggggc cacttcgtta agcatgtccc tgactcgcat gttttccctg 4561 accaaatccg ccagaaggcg ctcgccgccc agcgatagca gttcttgcaa ggaagcaaag 4621 tttttcaacg gtttgagacc gtccgccgta ggcatgcttt tgagcgtttg accaagcagt 4681 tccaggcggt cccacagctc ggtcacctgc tctacggcat ctcgatccag catatctcct 4741 cgtttcgcgg gttggggcgg ctttcgctgt acggcagtag tcggtgctcg tccagacggg 4801 ccagggtcat gtctttccac gggcgcaggg tcctcgtcag cgtagtctgg gtcacggtga 4861 aggggtgcgc tccgggctgc gcgctggcca gggtgcgctt gaggctggtc ctgctggtgc 4921 tgaagcgctg ccggtcttcg ccctgcgcgt cggccaggta gcatttgacc atggtgtcat 4981 agtccagccc ctccgcggcg tggcccttgg cgcgcagctt gcccttggag gaggcgccgc 5041 acgaggggca gtgcagactt ttgagggcgt agagcttggg cgcgagaaat accgattccg 5101 gggagtaggc atccgcgccg caggccccgc agacggtctc gcattccacg agccaggtga 5161 gctctggccg ttcggggtca aaaaccaggt ttcccccatg ctttttgatg cgtttcttac 5221 ctctggtttc catgagccgg tgtccacgct cggtgacgaa aaggctgtcc gtgtccccgt 5281 atacagactt gagagggagt ttaaacgaat tcaatagctt gttgcatggg cggcgatata 5341 aaatgcaagg tgctgctcaa aaaatcaggc aaagcctcgc gcaaaaaaga aagcacatcg 5401 tagtcatgct catgcagata aaggcaggta agctccggaa ccaccacaga aaaagacacc 5461 atttttctct caaacatgtc tgcgggtttc tgcataaaca caaaataaaa taacaaaaaa 5521 acatttaaac attagaagcc tgtcttacaa caggaaaaac aacccttata agcataagac 5581 ggactacggc catgccggcg tgaccgtaaa aaaactggtc accgtgatta aaaagcacca 5641 ccgacagctc ctcggtcatg tccggagtca taatgtaaga ctcggtaaac acatcaggtt 5701 gattcacatc ggtcagtgct aaaaagcgac cgaaatagcc cgggggaata catacccgca 5761 ggcgtagaga caacattaca gcccccatag gaggtataac aaaattaata ggagagaaaa 5821 acacataaac acctgaaaaa ccctcctgcc taggcaaaat agcaccctcc cgctccagaa 5881 caacatacag cgcttccaca gcggcagcca taacagtcag ccttaccagt aaaaaagaaa 5941 acctattaaa aaaacaccac tcgacacggc accagctcaa tcagtcacag tgtaaaaaag 6001 ggccaagtgc agagcgagta tatataggac taaaaaatga cgtaacggtt aaagtccaca 6061 aaaaacaccc agaaaaccgc acgcgaacct acgcccagaa acgaaagcca aaaaacccac 6121 aacttcctca aatcgtcact tccgttttcc cacgttacgt cacttcccat tttaagaaaa 6181 ctacaattcc caacacatac aagttactcc gccctaaaac ctacgtcacc cgccccgttc 6241 ccacgccccg cgccacgtca caaactccac cccctcatta tcatattggc ttcaatccaa 6301 aataaggtat attattgatg atgttaatta acatgcatgg atccatatgc ggtgtgaaat 6361 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac 6421 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6481 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6541 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6601 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6661 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6721 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6781 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6841 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6901 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6961 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 7021 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 7081 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 7141 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 7201 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 7261 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7321 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7381 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7441 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7501 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7561 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7621 gccagttaat agtttgcgca acgttgttgc cattgctgca gccatgagat tatcaaaaag 7681 gatcttcacc tagatccttt tcacgtagaa agccagtccg cagaaacggt gctgaccccg 7741 gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca 7801 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag 7861 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa 7921 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagct ctgatcaaga 7981 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 8041 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 8101 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 8161 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac 8221 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 8281 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 8341 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 8401 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 8461 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 8521 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 8581 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 8641 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 8701 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 8761 catcgccttc tatcgccttc ttgacgagtt cttctgaatt ttgttaaaat ttttgttaaa 8821 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat 8881 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg 8941 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac 9001 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta 9061 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag 9121 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg 9181 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag 9241 gatcgaatta attcttaatt aacatcatca ataatatacc ttattttgga ttgaagccaa 9301 tatgataatg agggggtgga gtttgtgacg tggcgcgggg cgtgggaacg gggcgggtga 9361 cgtagtagtg tggcggaagt gtgatgttgc aagtgtggcg gaacacatgt aagcgacgga 9421 tgtggcaaaa gtgacgtttt tggtgtgcgc cggtgtacac aggaagtgac aattttcgcg 9481 cggttttagg cggatgttgt agtaaatttg ggcgtaaccg agtaagattt ggccattttc 9541 gcgggaaaac tgaataagag gaagtgaaat ctgaataatt ttgtgttact catagcgcgt //