LOCUS       sequence-502423-        7362 bp    DNA     circular SYN 10-MAR-2026
DEFINITION  Non-FRET aCatenin 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 7362)
  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 7362)
  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_aCatTS_indicator
FEATURES             Location/Qualifiers
     source          1..7362
                     /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     1807..1828
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2048..2069)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3287..3997
                     /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(3432..3450)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3700..3719
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4150..4271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4187..4206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4241..4260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4278..4733)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4415..4436)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4627..4646
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4760..4864
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4866..5223
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4866..5057
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4889..4909)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5074..5209
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5258..6052
                     /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             5258..6052
                     /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(5312..5331)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5922..5941
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6240..6259)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6284..6331
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6660..7248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7149..7168
                     /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 cgcgccgaaa tgactgccgt ccacgcaggc aacataaact tcaaatggga
      661 ccccaaaagt ctggagatta ggactctggc tgttgaaaga ctgttggagc ctcttgttac
      721 acaggttaca accctggtaa acaccaatag taaagggccg tctaataaaa agagaggccg
      781 ttctaagaag gcccatgttt tggctgcatc tgttgaacaa gcaactgaga atttcttgga
      841 aaagggggat aaaattgcaa aagagagcca gtttctcaag gaggagcttg tggttgctgt
      901 agaagatgtt cgaaaacaag gagatttgat gaagagcgct gctggggagt ttgcagatga
      961 tccatgctct tctgtgaagc gaggcaacat ggtccgggca gctcgagctt tgctctctgc
     1021 cgtcacccgg ctgctgattc tggctgacat ggcagatgtc tacaaattac ttgttcagct
     1081 gaaagttgtg gaagatggta tattgaaact gaggaatgct ggcaatgaac aagacttagg
     1141 gatacagtac aaagctctga aaccagaagt agataagctg aacatcatgg cagcaaaaag
     1201 acaacaggaa ctaaaagatg tgggcaatcg tgatcagatg gctgcagcta gaggaatcct
     1261 gcagaaaaat gttccaatcc tctatactgc atcccaggca tgtctacaac accctgatgt
     1321 tgcagcctat aaggccaacc gggacttgat atacaagcag ctgcagcagg ccgtcacagg
     1381 aatttctaat gcagcccagg ctactgcatc agatgatgct gcccagcacc agggtggcag
     1441 tggcggagag ctggcatacg ctctcaacaa ctttgataaa caaatcattg tggacccctt
     1501 gagcttcagc gaggagcgct ttaggccgtc cctggaggag cgcctggaaa gcattattag
     1561 tggggaattc ggtggttctg gtggtggttc tggttccggt aacgtctata tcaaggccga
     1621 caagcagaag aacggcatca aggcgaactt caagatccgc cacaacatcg aggacggcgg
     1681 cgtgcagctc gcctaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1741 gcccgacaac cactacctga gcgttcagaa taaactttcg aaagacccca acgagaagcg
     1801 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1861 gctgtacaag gaattcgggc caggtggtgc agggccaggt ggtgcagggc caggtggtgc
     1921 agggccaggt ggtgcagggc ccggtggtgc aggcccaggt ggtgcaggtc caggtggtgc
     1981 aggtccaggt ggtgctggat ccatggtgag caagggcgag gagctgttca ccggggtggt
     2041 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2101 gggtgagggc gatgccacct acggcaagct gaccctgaag ttcttctgca ccaccggcaa
     2161 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2221 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2281 catccaggag cgcaccatct tcttcaagga cgacggctac tacaagaccc gcgccgaggt
     2341 gaagttcgag ggcgacaccc tggtgaaccg catcgtgctg aagggcatcg acttcaagga
     2401 ggacggcaac atcctggggc acaagctgga gtacaacagc ggccggggta gcggctgcgg
     2461 tagccaattg gactctgact tcgagactga agactttgat gtcagaagca ggaccagtgt
     2521 ccagacagaa gatgaccagc tgatagctgg ccagagcgcc cgggcgatca tggctcagct
     2581 tcctcaggag caaaaagcaa agattgcaga acaggtggcc agtttccagg aagagaagag
     2641 caagctggat gctgaagtgt ccaagtggga tgacagtggc aatgacatca ttgtgctggc
     2701 caagcagatg tgcatgatca tgatggagat gaccgacttc acccgaggca aaggtccact
     2761 caaaaacacc tctgacgtga tcagtgctgc caagaaaatt gcggaggcgg gatccaggat
     2821 ggacaagctc ggccggacca tcgcagacca ttgcccagac tcggcatgca agcaggacct
     2881 gctggcctac cttcagcgca tcgctctcta ctgccaccag ctcaacatct gcagcaaagt
     2941 caaggctgaa gtacagaacc ttggtggaga gctggtcgtc tctggggtgg acagcgccat
     3001 gtccctgatc caggcggcca agaacttgat gaatgctgtt gtgcagacag tgaaggcgtc
     3061 ctacgtggct tccaccaaat accagaagtc acaggggatg gcttccttga accttcctgc
     3121 tgtgtcatgg aagatgaagg cccctgagaa gaagccgttg gtgaagagag agaagcagga
     3181 tgagacgcag accaagatta aacgagcttc tcagaagaaa cacgtgaacc cagtgcaggc
     3241 cctgagcgag ttcaaagcca tggacagcat cagcggccgg ggaaccatgg tgagcaaggg
     3301 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     3361 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     3421 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     3481 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     3541 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     3601 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     3661 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     3721 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     3781 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     3841 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     3901 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     3961 ccactccacc ggcggcatgg acgagctgta caagtaagcg gccgcggtac cgcgggcccg
     4021 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     4081 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     4141 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4201 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4261 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     4321 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     4381 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     4441 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     4501 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     4561 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     4621 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4681 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     4741 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4801 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4861 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4921 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4981 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5041 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5101 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5161 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5221 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5281 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5341 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5401 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     5461 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5521 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5581 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5641 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     5701 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     5761 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     5821 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     5881 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     5941 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6001 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6061 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6121 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6181 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     6241 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     6301 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     6361 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     6421 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     6481 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     6541 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6601 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6661 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6721 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6781 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6841 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6901 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6961 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7021 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7081 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7141 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7201 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7261 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7321 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
