LOCUS       sequence-246088-        8240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET-based tension sensor (TS) technology to develop a probe for 
            molecular-level tension across alphaE-catenin..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8240)
  AUTHORS   Acharya BR, Wu SK, Lieu ZZ, Parton RG, Grill SW, Bershadsky AD, 
            Gomez GA, Yap AS
  TITLE     Mammalian Diaphanous 1 Mediates a Pathway for E-cadherin to 
            Stabilize Epithelial Barriers through Junctional Contractility.
  JOURNAL   Cell Rep. 2017 Mar 21;18(12):2854-2867. doi: 
            10.1016/j.celrep.2017.02.078.
  PUBMED    28329679
REFERENCE   2  (bases 1 to 8240)
  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"; doi: 
            "10.1016/j.celrep.2017.02"; journalName: "Cell Rep"; date: 
            "2017-03-21- 21"; volume: "18"; issue: "12"; pages: "2854-2867"
COMMENT     Sequence Label: aCat TS WT (1-697 mTFP1-TSmod-Venus 698-907)
FEATURES             Location/Qualifiers
     source          1..8240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     501..520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        775..1078
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1079..1282
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1233..1253
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3437..4144
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     CDS             join(4265..4267,4268..4270,4271..4981)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4310..4331)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4571..4590)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4918..4939
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5742..5863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5779..5798)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5833..5852
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5870..6325)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6007..6028)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6219..6238
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6352..6456
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6458..6815
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6458..6649
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6481..6501)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6666..6801
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6850..7644
                     /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             6850..7644
                     /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(6904..6923)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7514..7533
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7832..7851)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7876..7923
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
       61 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      121 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      181 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      241 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      301 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      361 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      421 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      481 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      541 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      601 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      661 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      721 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      781 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      841 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      901 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      961 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     1021 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     1081 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     1141 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1201 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1261 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
     1321 tcagatctcg agcgccacca tgactgccgt ccacgcaggc aacataaact tcaaatggga
     1381 ccccaaaagt ctggagatta ggactctggc tgttgaaaga ctgttggagc ctcttgttac
     1441 acaggttaca accctggtaa acaccaatag taaagggccg tctaataaaa agagaggccg
     1501 ttctaagaag gcccatgttt tggctgcatc tgttgaacaa gcaactgaga atttcttgga
     1561 aaagggggat aaaattgcaa aagagagcca gtttctcaag gaggagcttg tggttgctgt
     1621 agaagatgtt cgaaaacaag gagatttgat gaagagcgct gctggggagt ttgcagatga
     1681 tccatgctct tctgtgaagc gaggcaacat ggtccgggca gctcgagctt tgctctctgc
     1741 cgtcacccgg ctgctgattc tggctgacat ggcagatgtc tacaaattac ttgttcagct
     1801 gaaagttgtg gaagatggta tattgaaact gaggaatgct ggcaatgaac aagacttagg
     1861 gatacagtac aaagctctga aaccagaagt agataagctg aacatcatgg cagcaaaaag
     1921 acaacaggaa ctaaaagatg tgggcaatcg tgatcagatg gctgcagcta gaggaatcct
     1981 gcagaaaaat gttccaatcc tctatactgc atcccaggca tgtctacaac accctgatgt
     2041 tgcagcctat aaggccaacc gggacttgat atacaagcag ctgcagcagg ccgtcacagg
     2101 aatttctaat gcagcccagg ctactgcatc agatgatgct gcccagcacc agggtggcag
     2161 tggcggagag ctggcatacg ctctcaacaa ctttgataaa caaatcattg tggacccctt
     2221 gagcttcagc gaggagcgct ttaggccgtc cctggaggag cgcctggaaa gcattattag
     2281 tggggctgcc ctgatggctg actcatcctg cacacgtgat gaccgtcggg agcgcattgt
     2341 ggcagagtgt aatgctgtcc gccaggctct acaggacctg ctctcagaat acatgggcaa
     2401 cgctggacgt aaagaaagaa gtgatgcact caactctgca atagataaaa tgaccaagaa
     2461 gaccagggac ttgcgtagac agcttcgcaa agctgtcatg gaccacgtat cagattcttt
     2521 cctggagacc aatgtccctc tattagtact gattgaagct gcaaagaatg gaaatgagaa
     2581 agaagttaag gaatatgccc aagtttttcg tgaacatgcc aacaaactga ttgaggttgc
     2641 caacctggcc tgttcaatct ccaacaatga agaaggcgtg aagcttgtcc gaatgtctgc
     2701 aagccagtta gaagcgctgt gtcctcaggt tatcaatgct gcattggcgt tggcagcaaa
     2761 gccgcagagt aaactggccc aagagaacat ggatcttttt aaagagcaat gggaaaagca
     2821 agtccgtgtt ctcacagatg ctgttgatga cattacttcc atcgatgact tcttggctgt
     2881 ctcagagaac cacattttag aagatgtgaa caagtgtgtt attgctctcc aagagaaaga
     2941 cgtggatggg ctggatcgca ctgccggtgc catccgaggc cgggcagccc gagtcattca
     3001 tgtagtcacc tcagagatgg acaactacga gccaggagtt tacacagaga aggttctgga
     3061 agccaccaag ctcctctcca acacagtcat gccgcgcttc actgagcagg tggaagcagc
     3121 tgtggaagcc ctcagctcgg acccggccca gcccatggat gagaatgagt tcatcgacgc
     3181 ctcccgtctg gtgtacgacg gcatccggga catccggaaa gcagtgctga tgatcaggac
     3241 ccccgaggag ttggacgact ctgacttcga gactgaagac tttgatgtca gaagcaggac
     3301 cagtgtccag acagaagatg accagctgat agctggccag agcgcccggg cgatcatggc
     3361 tcagcttcct caggagcaaa aagcaaagat tgcagaacag gtggccagtt tccaggaaga
     3421 gaagagcaag ctcgagatgg tgagcaaggg cgaggagacc acaatgggcg taatcaagcc
     3481 cgacatgaag atcaagctga agatggaggg caacgtgaat ggccacgcct tcgtgatcga
     3541 gggcgagggc gagggcaagc cctacgacgg caccaacacc atcaacctgg aggtgaagga
     3601 gggagccccc ctgcccttct cctacgacat tctgaccacc gcgttcgcct acggcaacag
     3661 ggccttcacc aagtaccccg acgacatccc caactacttc aagcagtcct tccccgaggg
     3721 ctactcttgg gagcgcacca tgaccttcga ggacaagggc atcgtgaagg tgaagtccga
     3781 catctccatg gaggaggact ccttcatcta cgagatacac ctcaagggcg agaacttccc
     3841 ccccaacggc cccgtgatgc agaagaagac caccggctgg gacgcctcca ccgagaggat
     3901 gtacgtgcgc gacggcgtgc tgaagggcga cgtcaagcac aagctgctgc tggagggcgg
     3961 cggccaccac cgcgttgact tcaagaccat ctacagggcc aagaaggcgg tgaagctgcc
     4021 cgactatcac tttgtggacc accgcatcga gatcctgaac cacgacaagg actacaacaa
     4081 ggtgaccgtt tacgagagcg ccgtggcccg caactccacc gacggcatgg acgagctgta
     4141 caaggggcca ggtggtgcag ggccaggtgg tgcagggcca ggtggtgcag ggccaggtgg
     4201 tgcagggccc ggtggtgcag gtccaggtgg tgcaggtcca ggtggtgcag gtccaggtgg
     4261 tgctatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4321 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     4381 ctacggcaag ctgaccctga agctgatctg caccaccggc aagctgcccg tgccctggcc
     4441 caccctcgtg accaccctgg gctacggcct gcagtgcttc gcccgctacc ccgaccacat
     4501 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     4561 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     4621 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     4681 gcacaagctg gagtacaact acaacagcca caacgtctat atcaccgccg acaagcagaa
     4741 gaacggcatc aaggccaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct
     4801 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     4861 ccactacctg agctaccagt ccaaactgag caaagacccc aacgagaagc gcgatcacat
     4921 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     4981 ggcggccgcg ctggatgctg aagtgtccaa gtgggatgac agtggcaatg acatcattgt
     5041 gctggccaag cagatgtgca tgatcatgat ggagatgacc gacttcaccc gaggcaaagg
     5101 tccactcaaa aacacctctg acgtgatcag tgctgccaag aaaattgcgg aggcgggatc
     5161 caggatggac aagctcggcc ggaccatcgc agaccattgc ccagactcgg catgcaagca
     5221 ggacctgctg gcctaccttc agcgcatcgc tctctactgc caccagctca acatctgcag
     5281 caaagtcaag gctgaagtac agaaccttgg tggagagctg gtcgtctctg gggtggacag
     5341 cgccatgtcc ctgatccagg cggccaagaa cttgatgaat gctgttgtgc agacagtgaa
     5401 ggcgtcctac gtggcttcca ccaaatacca gaagtcacag gggatggctt ccttgaacct
     5461 tcctgctgtg tcatggaaga tgaaggcccc tgagaagaag ccgttggtga agagagagaa
     5521 gcaggatgag acgcagacca agattaaacg agcttctcag aagaaacacg tgaacccagt
     5581 gcaggccctg agcgagttca aagccatgga cagcatctga gcggccgcga ctctagatca
     5641 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     5701 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     5761 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5821 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     5881 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5941 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     6001 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     6061 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     6121 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     6181 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     6241 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     6301 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6361 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6421 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     6481 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6541 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     6601 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     6661 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6721 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6781 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     6841 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6901 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6961 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     7021 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     7081 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     7141 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     7201 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     7261 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     7321 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     7381 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     7441 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     7501 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     7561 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     7621 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     7681 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     7741 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     7801 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     7861 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     7921 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     7981 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     8041 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     8101 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     8161 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     8221 cagaccccgt agaaaagatc
//
