LOCUS       sequence-502419-        8238 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  Non-FRET aActinin tension indicator. cpEGFP with elastic linker is 
            inserted at the hinge with C-term mCherry..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8238)
  AUTHORS   Fujiwara K, Fujiki K, Akama TO, Shirahige K, Shiojima I, Nakamura T,
            Hirai M
  TITLE     Molecular tension indicators reveal unexpectedly complex regulation 
            of tension in live mouse organs.
  JOURNAL   Commun Biol. 2026 Feb 19. doi: 10.1038/s42003-026-09746-0.
  PUBMED    41714452
REFERENCE   2  (bases 1 to 8238)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 10, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol. 2026 Feb 19. doi: 10.1038/s42003-026-09746-0."
COMMENT     Sequence Label: pCMV_aActTS_indicator
FEATURES             Location/Qualifiers
     source          1..8238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     primer_bind     1735..1756
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1976..1997)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             4163..4873
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4308..4326)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4576..4595
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5026..5147
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5063..5082)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5117..5136
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5154..5609)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5291..5312)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5503..5522
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5636..5740
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5742..6099
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5742..5933
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5765..5785)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5950..6085
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6134..6928
                     /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             6134..6928
                     /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(6188..6207)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6798..6817
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7116..7135)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7160..7207
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7536..8124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8025..8044
                     /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 gctaggagcg
      601 ctgatatcgg cgcgccccta gcatggacca ttatgattct cagcaaacca acgattacat
      661 gcagccagaa gaggactggg accgggacct gctcctggac ccggcctggg agaagcagca
      721 gagaaagaca ttcacggcat ggtgtaactc ccacctccgg aaggcgggga cacagatcga
      781 gaacatcgaa gaggacttcc gggatggcct gaagctcatg ctgctgctgg aggtcatctc
      841 aggtgaacgc ttggccaagc cagagcgagg caagatgaga gtgcacaaga tctccaacgt
      901 caacaaggcc ctggatttca tagccagcaa aggcgtcaaa ctggtgtcca tcggagccga
      961 agaaatcgtg gatgggaatg tgaagatgac cctgggcatg atctggacca tcatcctgcg
     1021 ctttgccatc caggacatct ccgtggaaga gacttcagcc aaggaagggc tgctcctgtg
     1081 gtgtcagaga aagacagccc cttacaaaaa tgtcaacatc cagaacttcc acataagctg
     1141 gaaggatggc ctcggcttct gtgctttgat ccaccgacac cggcccgagc tgattgacta
     1201 cgggaagctg cggaaggatg atccactcac aaatctgaat acggcttttg acgtggcaga
     1261 gaagtacctg gacatcccca agatgctgga tgccgaagac atcgttggaa ctgcccgacc
     1321 ggatgagaaa gccatcatga cttacgtgtc tagcttctac cacgccttct ctggagccca
     1381 gaaggcggag acagcagcca atcgcatctg caaggtgttg gccgtcaacc aggagaacga
     1441 gcagcttatg gaagactacg agaagctggc cagtgatctg ttggagtgga tccgccgcac
     1501 aatcccgtgg ctggagaacc ggaccggtaa cgtctatatc aaggccgaca agcagaagaa
     1561 cggcatcaag gcgaacttca agatccgcca caacatcgag gacggcggcg tgcagctcgc
     1621 ctaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1681 ctacctgagc gttcagaata aactttcgaa agaccccaac gagaagcgcg atcacatggt
     1741 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagga
     1801 attcgggcca ggtggtgcag ggccaggtgg tgcagggcca ggtggtgcag ggccaggtgg
     1861 tgcagggccc ggtggtgcag gcccaggtgg tgcaggtcca ggtggtgcag gtccaggtgg
     1921 tgctggatcc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1981 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga
     2041 tgccacctac ggcaagctga ccctgaagtt cttctgcacc accggcaagc tgcccgtgcc
     2101 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2161 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg
     2221 caccatcttc ttcaaggacg acggctacta caagacccgc gccgaggtga agttcgaggg
     2281 cgacaccctg gtgaaccgca tcgtgctgaa gggcatcgac ttcaaggagg acggcaacat
     2341 cctggggcac aagctggagt acaacagcgg ccgggtgccc gagaacacca tgcatgccat
     2401 gcaacagaag ctggaggact tccgggacta ccggcgcctg cacaagccgc ccaaggtgca
     2461 ggagaagtgc cagctggaga tcaacttcaa cacgctgcag accaagctgc ggctcagcaa
     2521 ccggcctgcc ttcatgccct ctgagggcag gatggtctcg gacatcaaca atgcctgggg
     2581 ctgcctggag caggtggaga agggctatga ggagtggttg ctgaatgaga tccggaggct
     2641 ggagcgactg gaccacctgg cagagaagtt ccggcagaag gcctccatcc acgaggcctg
     2701 gactgacggc aaagaggcca tgctgcgaca gaaggactat gagaccgcca ccctctcgga
     2761 gatcaaggcc ttgctcaaga agcatgaggc cttcgagagt gacctggctg cccaccagga
     2821 ccgtgtggag cagattgccg ccatcgcaca ggagctcaat gagctggact attatgactc
     2881 acccagtgtc aacgcccgtt gccaaaagat ctgtgaccag tgggacaatc tgggggccct
     2941 aactcagaag cgaagggaag ctctggagcg gaccgagaaa ctgctggaga ccattgacca
     3001 gctgtacttg gagtatgcca agcgggctgc acccttcaac aactggatgg agggggccat
     3061 ggaggacctg caggacacct tcattgtgca caccattgag gagatccagg gactgaccac
     3121 agcccatgag cagttcaagg ccaccctccc tgatgccgac aaggagcgcc tggccatcct
     3181 gggcatccac aatgaggtgt ccaagattgt ccagacctac cacgtcaata tggcgggcac
     3241 caacccctac acaaccatca cgcctcagga gatcaatggc aaatgggacc acgtgcggca
     3301 gctggtgcct cggagggacc aagctctgac ggaggagcat gcccgacagc agcacaatga
     3361 gaggctacgc aagcagtttg gagcccaggc caatgtcatc gggccctgga tccagaccaa
     3421 gatggaggag atcgggagga tctccattga gatgcatggg accctggagg accagctcag
     3481 ccacctgcgg cagtatgaga agagcatcgt caactacaag ccaaagattg atcagctgga
     3541 gggcgaccac cagctcatcc aggaggcgct catcttcgac aacaagcaca ccaactacac
     3601 catggagcac atccgtgtgg gctgggagca gctgctcacc accatcgcca ggaccatcaa
     3661 tgaggtagag aaccagatcc tgacccggga tgccaagggc atcagccagg agcagatgaa
     3721 tgagttccgg gcctccttca accactttga ccgggatcac tccggcacac tgggtcccga
     3781 ggagttcaaa gcctgcctca tcagcttggg ttatgatatt ggcaacgacc cccagggaga
     3841 agcagaattt gcccgcatca tgagcattgt ggaccccaac cgcctggggg tagtgacatt
     3901 ccaggccttc attgacttca tgtcccgcga gacagccgac acagatacag cagaccaagt
     3961 catggcttcc ttcaagatcc tggctgggga caagaactac attaccatgg acgagctgcg
     4021 ccgcgagctg ccacccgacc aggctgagta ctgcatcgcg cggatggccc cctacaccgg
     4081 ccccgactcc gtgccaggtg ctctggacta catgtccttc tccacggcgc tgtacggcga
     4141 gagtgacctc actagtggaa ccatggtgag caagggcgag gaggataaca tggccatcat
     4201 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     4261 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     4321 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     4381 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     4441 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     4501 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     4561 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     4621 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     4681 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     4741 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     4801 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     4861 gctgtacaag taagcggccg cggtaccgcg ggcccgggat ccaccggatc tagataactg
     4921 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     4981 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     5041 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5101 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     5161 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5221 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5281 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5341 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5401 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5461 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     5521 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     5581 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     5641 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5701 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     5761 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     5821 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5881 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5941 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     6001 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     6061 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     6121 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     6181 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     6241 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     6301 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     6361 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     6421 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     6481 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     6541 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     6601 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     6661 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     6721 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     6781 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     6841 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6901 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     6961 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     7021 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     7081 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     7141 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     7201 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     7261 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     7321 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     7381 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7441 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7501 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7561 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7621 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7681 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7741 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7801 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7861 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7921 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7981 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8041 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8101 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8161 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8221 cgtattaccg ccatgcat
//
