LOCUS       sequence-243833-        9363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Myofilament localised red genetically encoded calcium sensor in 
            mammalian expression vector used for AdEasy adenoviral DNA 
            recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9363)
  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 9363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Circ Res. 
            2019 Feb 8. doi: 10.1161/CIRCRESAHA.118.314600."
COMMENT     Sequence Label: pShuttle CMV RGECO-TnI
FEATURES             Location/Qualifiers
     source          1..9363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             633..689
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     1385..1404
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2663..2784
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2700..2719)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2754..2773
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   5975..6077
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(6166..6183)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6337..6925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6417..6436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             7757..8551
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             7757..8551
                     /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(7811..7830)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8421..8440
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(8599..8979)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8661..8682)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8873..8892
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   9017..9119
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     9207..9357
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagagat ctggtaccgt
      601 cgacgcggcc gcccaccatg gtagactcat cacgtcgtaa gtggaataag gcaggtcacg
      661 cagtcagagc tataggtcgg ctgagctcac ccgtggtttc cgagcggatg taccccgagg
      721 acggcgccct caagagcgag atcaagaagg ggctgaggct gaaggacggc ggccactacg
      781 ccgccgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
      841 tcgtagacat caagttggac atcgtgtccc acaacgagga ctacaccatc gtggaacagt
      901 gcgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctatac aagggaggta
      961 caggcgggag tctggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     1021 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     1081 agggcgaggg ccgcccctac gaggcctttc agaccgctaa gctgaaggtg accaagggtg
     1141 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     1201 acattaagca cccagccgac atccccgact acttcaagct gtccttcccc gagggcttca
     1261 ggtgggagcg cgtgatgaac ttcgaggacg gcggcattat tcacgttaac caggactcct
     1321 ccctgcagga cggcgtattc atctacaagg tgaagctgcg cggcaccaac ttcccccccg
     1381 acggccccgt aatgcagaag aagaccatgg gctgggaggc tacgcgtgac caactgactg
     1441 aggagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac ggggatggga
     1501 cgataacaac caaggagctg gggacggtga tgcggtctct ggggcagaac cccacagaag
     1561 cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca ttcgacttcc
     1621 ctgagttcct gacgatgatg gcaagaaaaa tgaatgacac agacagtgaa gaggaaatta
     1681 gagaagcgtt ccgcgtgttt gataaggacg gcaatggcta catcggcgca gcagagcttc
     1741 gccacgtgat gacagacctt ggagagaagt taacagatga ggaggttgat gaaatgatca
     1801 gggtagcaga catcgatggg gatggtcagg taaactacga agagtttgtc caaatgatga
     1861 cagcgaagca tatgctcgag atggcggatg ggagcagcga tgcggctagg gaacctcgcc
     1921 ctgcaccagc cccaatcaga cgccgctcct ccaactaccg cgcttatgcc acggagccgc
     1981 acgccaagaa aaaatctaag atctccgctt caagaaaatt gcagctgaag actctgctgc
     2041 tgcagattgc aaagcaagag ctggagcgag aggcggagga gcggcgcggg gagaaggggc
     2101 gcgctctgag cacccgctgc cagccgctgg agttgaccgg gctgggcttc gcggagctgc
     2161 aggacttgtg ccgacagctc cacgcccgtg tggacaaggt ggatgaagag agatacgaca
     2221 tagaggcaaa agtcaccaag aacatcacgg agattgcaga tctgactcag aagatctttg
     2281 accttcgagg caagtttaag cggcccaccc tgcggagagt gaggatctct gcagatgcca
     2341 tgatgcaggc gctgctgggg gcccgggcta aggagtccct ggacctgcgg gcccacctca
     2401 agcaggtgaa gaaggaggac accgagaagg aaaaccggga ggtgggagac tggcgcaaga
     2461 acatcgatgc actgagtgga atggagggcc gcaagaaaaa gtttgagagc tgaaagcttc
     2521 tagataagat atccgatcca ccggatctag ataactgatc ataatcagcc ataccacatt
     2581 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2641 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2701 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2761 gtccaaactc atcaatgtat cttaacgcgg atctgggcgt ggttaagggt gggaaagaat
     2821 atataaggtg ggggtcttat gtagttttgt atctgttttg cagcagccgc cgccgccatg
     2881 agcaccaact cgtttgatgg aagcattgtg agctcatatt tgacaacgcg catgccccca
     2941 tgggccgggg tgcgtcagaa tgtgatgggc tccagcattg atggtcgccc cgtcctgccc
     3001 gcaaactcta ctaccttgac ctacgagacc gtgtctggaa cgccgttgga gactgcagcc
     3061 tccgccgccg cttcagccgc tgcagccacc gcccgcggga ttgtgactga ctttgctttc
     3121 ctgagcccgc ttgcaagcag tgcagcttcc cgttcatccg cccgcgatga caagttgacg
     3181 gctcttttgg cacaattgga ttctttgacc cgggaactta atgtcgtttc tcagcagctg
     3241 ttggatctgc gccagcaggt ttctgccctg aaggcttcct cccctcccaa tgcggtttaa
     3301 aacataaata aaaaaccaga ctctgtttgg atttggatca agcaagtgtc ttgctgtctt
     3361 tatttagggg ttttgcgcgc gcggtaggcc cgggaccagc ggtctcggtc gttgagggtc
     3421 ctgtgtattt tttccaggac gtggtaaagg tgactctgga tgttcagata catgggcata
     3481 agcccgtctc tggggtggag gtagcaccac tgcagagctt catgctgcgg ggtggtgttg
     3541 tagatgatcc agtcgtagca ggagcgctgg gcgtggtgcc taaaaatgtc tttcagtagc
     3601 aagctgattg ccaggggcag gcccttggtg taagtgttta caaagcggtt aagctgggat
     3661 gggtgcatac gtggggatat gagatgcatc ttggactgta tttttaggtt ggctatgttc
     3721 ccagccatat ccctccgggg attcatgttg tgcagaacca ccagcacagt gtatccggtg
     3781 cacttgggaa atttgtcatg tagcttagaa ggaaatgcgt ggaagaactt ggagacgccc
     3841 ttgtgacctc caagattttc catgcattcg tccataatga tggcaatggg cccacgggcg
     3901 gcggcctggg cgaagatatt tctgggatca ctaacgtcat agttgtgttc caggatgaga
     3961 tcgtcatagg ccatttttac aaagcgcggg cggagggtgc cagactgcgg tataatggtt
     4021 ccatccggcc caggggcgta gttaccctca cagatttgca tttcccacgc tttgagttca
     4081 gatgggggga tcatgtctac ctgcggggcg atgaagaaaa cggtttccgg ggtaggggag
     4141 atcagctggg aagaaagcag gttcctgagc agctgcgact taccgcagcc ggtgggcccg
     4201 taaatcacac ctattaccgg ctgcaactgg tagttaagag agctgcagct gccgtcatcc
     4261 ctgagcaggg gggccacttc gttaagcatg tccctgactc gcatgttttc cctgaccaaa
     4321 tccgccagaa ggcgctcgcc gcccagcgat agcagttctt gcaaggaagc aaagtttttc
     4381 aacggtttga gaccgtccgc cgtaggcatg cttttgagcg tttgaccaag cagttccagg
     4441 cggtcccaca gctcggtcac ctgctctacg gcatctcgat ccagcatatc tcctcgtttc
     4501 gcgggttggg gcggctttcg ctgtacggca gtagtcggtg ctcgtccaga cgggccaggg
     4561 tcatgtcttt ccacgggcgc agggtcctcg tcagcgtagt ctgggtcacg gtgaaggggt
     4621 gcgctccggg ctgcgcgctg gccagggtgc gcttgaggct ggtcctgctg gtgctgaagc
     4681 gctgccggtc ttcgccctgc gcgtcggcca ggtagcattt gaccatggtg tcatagtcca
     4741 gcccctccgc ggcgtggccc ttggcgcgca gcttgccctt ggaggaggcg ccgcacgagg
     4801 ggcagtgcag acttttgagg gcgtagagct tgggcgcgag aaataccgat tccggggagt
     4861 aggcatccgc gccgcaggcc ccgcagacgg tctcgcattc cacgagccag gtgagctctg
     4921 gccgttcggg gtcaaaaacc aggtttcccc catgcttttt gatgcgtttc ttacctctgg
     4981 tttccatgag ccggtgtcca cgctcggtga cgaaaaggct gtccgtgtcc ccgtatacag
     5041 acttgagagg gagtttaaac gaattcaata gcttgttgca tgggcggcga tataaaatgc
     5101 aaggtgctgc tcaaaaaatc aggcaaagcc tcgcgcaaaa aagaaagcac atcgtagtca
     5161 tgctcatgca gataaaggca ggtaagctcc ggaaccacca cagaaaaaga caccattttt
     5221 ctctcaaaca tgtctgcggg tttctgcata aacacaaaat aaaataacaa aaaaacattt
     5281 aaacattaga agcctgtctt acaacaggaa aaacaaccct tataagcata agacggacta
     5341 cggccatgcc ggcgtgaccg taaaaaaact ggtcaccgtg attaaaaagc accaccgaca
     5401 gctcctcggt catgtccgga gtcataatgt aagactcggt aaacacatca ggttgattca
     5461 catcggtcag tgctaaaaag cgaccgaaat agcccggggg aatacatacc cgcaggcgta
     5521 gagacaacat tacagccccc ataggaggta taacaaaatt aataggagag aaaaacacat
     5581 aaacacctga aaaaccctcc tgcctaggca aaatagcacc ctcccgctcc agaacaacat
     5641 acagcgcttc cacagcggca gccataacag tcagccttac cagtaaaaaa gaaaacctat
     5701 taaaaaaaca ccactcgaca cggcaccagc tcaatcagtc acagtgtaaa aaagggccaa
     5761 gtgcagagcg agtatatata ggactaaaaa atgacgtaac ggttaaagtc cacaaaaaac
     5821 acccagaaaa ccgcacgcga acctacgccc agaaacgaaa gccaaaaaac ccacaacttc
     5881 ctcaaatcgt cacttccgtt ttcccacgtt acgtcacttc ccattttaag aaaactacaa
     5941 ttcccaacac atacaagtta ctccgcccta aaacctacgt cacccgcccc gttcccacgc
     6001 cccgcgccac gtcacaaact ccaccccctc attatcatat tggcttcaat ccaaaataag
     6061 gtatattatt gatgatgtta attaacatgc atggatccat atgcggtgtg aaataccgca
     6121 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     6181 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6241 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6301 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6361 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6421 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6481 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6541 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6601 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6661 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6721 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6781 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6841 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6901 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6961 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7021 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7081 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7141 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7201 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7261 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7321 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7381 taatagtttg cgcaacgttg ttgccattgc tgcagccatg agattatcaa aaaggatctt
     7441 cacctagatc cttttcacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa
     7501 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc
     7561 ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc
     7621 ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat
     7681 ggctttcttg ccgccaagga tctgatggcg caggggatca agctctgatc aagagacagg
     7741 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     7801 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     7861 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     7921 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     7981 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     8041 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     8101 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     8161 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     8221 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     8281 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     8341 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     8401 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     8461 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     8521 cttctatcgc cttcttgacg agttcttctg aattttgtta aaatttttgt taaatcagct
     8581 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     8641 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     8701 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     8761 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     8821 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     8881 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     8941 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggatcga
     9001 attaattctt aattaacatc atcaataata taccttattt tggattgaag ccaatatgat
     9061 aatgaggggg tggagtttgt gacgtggcgc ggggcgtggg aacggggcgg gtgacgtagt
     9121 agtgtggcgg aagtgtgatg ttgcaagtgt ggcggaacac atgtaagcga cggatgtggc
     9181 aaaagtgacg tttttggtgt gcgccggtgt acacaggaag tgacaatttt cgcgcggttt
     9241 taggcggatg ttgtagtaaa tttgggcgta accgagtaag atttggccat tttcgcggga
     9301 aaactgaata agaggaagtg aaatctgaat aattttgtgt tactcatagc gcgtaatact
     9361 gta
//
