LOCUS       sequence-149113-        8204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET-based actinin tension sensors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8204)
  AUTHORS   Yamashita S, Tsuboi T, Ishinabe N, Kitaguchi T, Michiue T
  TITLE     Wide and high resolution tension measurement using FRET in embryo.
  JOURNAL   Sci Rep. 2016 Jun 23;6:28535. doi: 10.1038/srep28535.
  PUBMED    27335157
REFERENCE   2  (bases 1 to 8204)
  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: "Sci Rep."; 
            date: "2016-06-23"; volume: "6:28535. doi"; pages: " 
            10.1038/srep28535"
COMMENT     Sequence Label: ActTS-GR
FEATURES             Location/Qualifiers
     source          1..8204
                     /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"
     CDS             1503..2210
                     /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(1648..1666)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1916..1935
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(2343..2345,2346..2348,2349..3059)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2343..2345,2346..2348,2349..3059)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2388..2409)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2649..2668)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2996..3017
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4992..5113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5029..5048)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5083..5102
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5120..5575)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5257..5278)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5469..5488
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5602..5706
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5708..6065
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5708..5899
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5731..5751)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5916..6051
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6100..6894
                     /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             6100..6894
                     /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(6154..6173)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6764..6783
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7082..7101)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7126..7173
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7502..8090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7991..8010
                     /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 gctagcatgg
      601 accattatga ttctcagcaa accaacgatt acatgcagcc agaagaggac tgggaccggg
      661 acctgctcct ggacccggcc tgggagaagc agcagagaaa gacattcacg gcatggtgta
      721 actcccacct ccggaaggcg gggacacaga tcgagaacat cgaagaggac ttccgggatg
      781 gcctgaagct catgctgctg ctggaggtca tctcaggtga acgcttggcc aagccagagc
      841 gaggcaagat gagagtgcac aagatctcca acgtcaacaa ggccctggat ttcatagcca
      901 gcaaaggcgt caaactggtg tccatcggag ccgaagaaat cgtggatggg aatgtgaaga
      961 tgaccctggg catgatctgg accatcatcc tgcgctttgc catccaggac atctccgtgg
     1021 aagagacttc agccaaggaa gggctgctcc tgtggtgtca gagaaagaca gccccttaca
     1081 aaaatgtcaa catccagaac ttccacataa gctggaagga tggcctcggc ttctgtgctt
     1141 tgatccaccg acaccggccc gagctgattg actacgggaa gctgcggaag gatgatccac
     1201 tcacaaatct gaatacggct tttgacgtgg cagagaagta cctggacatc cccaagatgc
     1261 tggatgccga agacatcgtt ggaactgccc gaccggatga gaaagccatc atgacttacg
     1321 tgtctagctt ctaccacgcc ttctctggag cccagaaggc ggagacagca gccaatcgca
     1381 tctgcaaggt gttggccgtc aaccaggaga acgagcagct tatggaagac tacgagaagc
     1441 tggccagtga tctgttggag tggatccgcc gcacaatccc gtggctggag aaccggaccg
     1501 gtatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     1561 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     1621 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     1681 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     1741 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     1801 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tcccttcagg
     1861 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1921 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     1981 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2041 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2101 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2161 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag gaattcgggc
     2221 caggtggtgc agggccaggt ggtgcagggc caggtggtgc agggccaggt ggtgcagggc
     2281 ccggtggtgc aggcccaggt ggtgcaggtc caggtggtgc aggtccaggt ggtgctggat
     2341 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2401 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2461 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2521 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     2581 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2641 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2701 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2761 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     2821 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     2881 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2941 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3001 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaaga
     3061 gcggccgcgt gcccgagaac accatgcatg ccatgcaaca gaagctggag gacttccggg
     3121 actaccggcg cctgcacaag ccgcccaagg tgcaggagaa gtgccagctg gagatcaact
     3181 tcaacacgct gcagaccaag ctgcggctca gcaaccggcc tgccttcatg ccctctgagg
     3241 gcaggatggt ctcggacatc aacaatgcct ggggctgcct ggagcaggtg gagaagggct
     3301 atgaggagtg gttgctgaat gagatccgga ggctggagcg actggaccac ctggcagaga
     3361 agttccggca gaaggcctcc atccacgagg cctggactga cggcaaagag gccatgctgc
     3421 gacagaagga ctatgagacc gccaccctct cggagatcaa ggccctgctc aagaagcatg
     3481 aggccttcga gagtgacctg gctgcccacc aggaccgtgt ggagcagatt gccgccatcg
     3541 cacaggagct caatgagctg gactattatg actcacccag tgtcaacgcc cgttgccaaa
     3601 agatctgtga ccagtgggac aatctggggg ccctaactca gaagcgaagg gaagctctgg
     3661 agcggaccga gaaactgctg gagaccattg accagctgta cttggagtat gccaagcggg
     3721 ctgcaccctt caacaactgg atggaggggg ccatggagga cctgcaggac accttcattg
     3781 tgcacaccat tgaggagatc cagggactga ccacagccca tgagcagttc aaggccaccc
     3841 tccctgatgc cgacaaggag cgcctggcca tcctgggcat ccacaatgag gtgtccaaga
     3901 ttgtccagac ctaccacgtc aatatggcgg gcaccaaccc ctacacaacc atcacgcctc
     3961 aggagatcaa tggcaaatgg gaccacgtgc ggcagctggt gcctcggagg gaccaagctc
     4021 tgacggagga gcatgcccga cagcagcaca atgagaggct acgcaagcag tttggagccc
     4081 aggccaatgt catcgggccc tggatccaga ccaagatgga ggagatcggg aggatctcca
     4141 ttgagatgca tgggaccctg gaggaccagc tcagccacct gcggcagtat gagaagagca
     4201 tcgtcaacta caagccaaag attgatcagc tggagggcga ccaccagctc atccaggagg
     4261 cgctcatctt cgacaacaag cacaccaact acaccatgga gcacatccgt gtgggctggg
     4321 agcagctgct caccaccatc gccaggacca tcaatgaggt agagaaccag atcctgaccc
     4381 gggatgccaa gggcatcagc caggagcaga tgaatgagtt ccgggcctcc ttcaaccact
     4441 ttgaccggga tcactccggc acactgggtc ccgaggagtt caaagcctgc ctcatcagct
     4501 tgggttatga tattggcaac gacccccagg gagaagcaga atttgcccgc atcatgagca
     4561 ttgtggaccc caaccgcctg ggggtagtga cattccaggc cttcattgac ttcatgtccc
     4621 gcgagacagc cgacacagat acagcagacc aagtcatggc ttccttcaag atcctggctg
     4681 gggacaagaa ctacattacc atggacgagc tgcgccgcga gctgccaccc gaccaggctg
     4741 agtactgcat cgcgcggatg gccccctaca ccggccccga ctccgtgcca ggtgctctgg
     4801 actacatgtc cttctccacg gcgctgtacg gcgagagtga cctctaaggt accgcgggcc
     4861 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     4921 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     4981 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5041 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5101 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5161 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5221 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5281 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5341 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5401 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5461 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5521 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     5581 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5641 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5701 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5761 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5821 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5881 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5941 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6001 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     6061 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     6121 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6181 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6241 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     6301 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6361 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6421 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6481 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6541 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6601 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     6661 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6721 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6781 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6841 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6901 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6961 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     7021 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     7081 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     7141 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     7201 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     7261 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     7321 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     7381 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7441 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7501 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7561 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7621 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     7681 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7741 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7801 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7861 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7921 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7981 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8041 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8101 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     8161 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
