LOCUS       sequence-502421-        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"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2438..2459
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2679..2700)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             4866..5576
                     /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(5011..5029)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5279..5298
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5729..5850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5766..5785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5820..5839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5857..6312)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5994..6015)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6206..6225
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6339..6443
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6445..6802
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6445..6636
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6468..6488)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6653..6788
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6837..7631
                     /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             6837..7631
                     /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(6891..6910)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7501..7520
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7819..7838)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7863..7910
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagga gcgctgatat cggcgcgccc
     1321 ctagcatgga ccattatgat tctcagcaaa ccaacgatta catgcagcca gaagaggact
     1381 gggaccggga cctgctcctg gacccggcct gggagaagca gcagagaaag acattcacgg
     1441 catggtgtaa ctcccacctc cggaaggcgg ggacacagat cgagaacatc gaagaggact
     1501 tccgggatgg cctgaagctc atgctgctgc tggaggtcat ctcaggtgaa cgcttggcca
     1561 agccagagcg aggcaagatg agagtgcaca agatctccaa cgtcaacaag gccctggatt
     1621 tcatagccag caaaggcgtc aaactggtgt ccatcggagc cgaagaaatc gtggatggga
     1681 atgtgaagat gaccctgggc atgatctgga ccatcatcct gcgctttgcc atccaggaca
     1741 tctccgtgga agagacttca gccaaggaag ggctgctcct gtggtgtcag agaaagacag
     1801 ccccttacaa aaatgtcaac atccagaact tccacataag ctggaaggat ggcctcggct
     1861 tctgtgcttt gatccaccga caccggcccg agctgattga ctacgggaag ctgcggaagg
     1921 atgatccact cacaaatctg aatacggctt ttgacgtggc agagaagtac ctggacatcc
     1981 ccaagatgct ggatgccgaa gacatcgttg gaactgcccg accggatgag aaagccatca
     2041 tgacttacgt gtctagcttc taccacgcct tctctggagc ccagaaggcg gagacagcag
     2101 ccaatcgcat ctgcaaggtg ttggccgtca accaggagaa cgagcagctt atggaagact
     2161 acgagaagct ggccagtgat ctgttggagt ggatccgccg cacaatcccg tggctggaga
     2221 accggaccgg taacgtctat atcaaggccg acaagcagaa gaacggcatc aaggcgaact
     2281 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga
     2341 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcgttcaga
     2401 ataaactttc gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2461 ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggaattcggg ccaggtggtg
     2521 cagggccagg tggtgcaggg ccaggtggtg cagggccagg tggtgcaggg cccggtggtg
     2581 caggcccagg tggtgcaggt ccaggtggtg caggtccagg tggtgctgga tccatggtga
     2641 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     2701 taaacggcca caagttcagc gtgtccggcg agggtgaggg cgatgccacc tacggcaagc
     2761 tgaccctgaa gttcttctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2821 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     2881 acttcttcaa gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg
     2941 acgacggcta ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     3001 gcatcgtgct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3061 agtacaacag cggccgggtg cccgagaaca ccatgcatgc catgcaacag aagctggagg
     3121 acttccggga ctaccggcgc ctgcacaagc cgcccaaggt gcaggagaag tgccagctgg
     3181 agatcaactt caacacgctg cagaccaagc tgcggctcag caaccggcct gccttcatgc
     3241 cctctgaggg caggatggtc tcggacatca acaatgcctg gggctgcctg gagcaggtgg
     3301 agaagggcta tgaggagtgg ttgctgaatg agatccggag gctggagcga ctggaccacc
     3361 tggcagagaa gttccggcag aaggcctcca tccacgaggc ctggactgac ggcaaagagg
     3421 ccatgctgcg acagaaggac tatgagaccg ccaccctctc ggagatcaag gccttgctca
     3481 agaagcatga ggccttcgag agtgacctgg ctgcccacca ggaccgtgtg gagcagattg
     3541 ccgccatcgc acaggagctc aatgagctgg actattatga ctcacccagt gtcaacgccc
     3601 gttgccaaaa gatctgtgac cagtgggaca atctgggggc cctaactcag aagcgaaggg
     3661 aagctctgga gcggaccgag aaactgctgg agaccattga ccagctgtac ttggagtatg
     3721 ccaagcgggc tgcacccttc aacaactgga tggagggggc catggaggac ctgcaggaca
     3781 ccttcattgt gcacaccatt gaggagatcc agggactgac cacagcccat gagcagttca
     3841 aggccaccct ccctgatgcc gacaaggagc gcctggccat cctgggcatc cacaatgagg
     3901 tgtccaagat tgtccagacc taccacgtca atatggcggg caccaacccc tacacaacca
     3961 tcacgcctca ggagatcaat ggcaaatggg accacgtgcg gcagctggtg cctcggaggg
     4021 accaagctct gacggaggag catgcccgac agcagcacaa tgagaggcta cgcaagcagt
     4081 ttggagccca ggccaatgtc atcgggccct ggatccagac caagatggag gagatcggga
     4141 ggatctccat tgagatgcat gggaccctgg aggaccagct cagccacctg cggcagtatg
     4201 agaagagcat cgtcaactac aagccaaaga ttgatcagct ggagggcgac caccagctca
     4261 tccaggaggc gctcatcttc gacaacaagc acaccaacta caccatggag cacatccgtg
     4321 tgggctggga gcagctgctc accaccatcg ccaggaccat caatgaggta gagaaccaga
     4381 tcctgacccg ggatgccaag ggcatcagcc aggagcagat gaatgagttc cgggcctcct
     4441 tcaaccactt tgaccgggat cactccggca cactgggtcc cgaggagttc aaagcctgcc
     4501 tcatcagctt gggttatgat attggcaacg acccccaggg agaagcagaa tttgcccgca
     4561 tcatgagcat tgtggacccc aaccgcctgg gggtagtgac attccaggcc ttcattgact
     4621 tcatgtcccg cgagacagcc gacacagata cagcagacca agtcatggct tccttcaaga
     4681 tcctggctgg ggacaagaac tacattacca tggacgagct gcgccgcgag ctgccacccg
     4741 accaggctga gtactgcatc gcgcggatgg ccccctacac cggccccgac tccgtgccag
     4801 gtgctctgga ctacatgtcc ttctccacgg cgctgtacgg cgagagtgac ctcactagtg
     4861 gaaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     4921 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     4981 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     5041 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
     5101 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     5161 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     5221 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     5281 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     5341 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     5401 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     5461 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     5521 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaagcgg
     5581 ccgcggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     5641 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     5701 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     5761 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5821 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     5881 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     5941 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     6001 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     6061 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     6121 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     6181 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     6241 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     6301 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6361 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6421 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     6481 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6541 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6601 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     6661 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6721 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6781 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     6841 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     6901 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     6961 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     7021 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     7081 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     7141 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     7201 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     7261 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     7321 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     7381 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     7441 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     7501 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     7561 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     7621 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     7681 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     7741 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     7801 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     7861 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     7921 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     7981 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     8041 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     8101 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     8161 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     8221 aaagatcaaa ggatcttc
//
