LOCUS       sequence-246082-        8208 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 8208)
  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 8208)
  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 TL1 (1-907 mTFP1-TSmod-Venus STOP)
FEATURES             Location/Qualifiers
     source          1..8208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      10..598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     499..518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        773..1076
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1077..1280
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1231..1251
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      4033..4042
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4039..4746
                     /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(4867..4869,4870..4872,4873..5586)
                     /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(4912..4933)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5173..5192)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5520..5541
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5708..5829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5745..5764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5799..5818
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5836..6291)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5973..5994)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6185..6204
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6318..6422
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6424..6781
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6424..6615
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6447..6467)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6632..6767
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6816..7610
                     /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             6816..7610
                     /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(6870..6889)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7480..7499
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7798..7817)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7842..7889
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
       61 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      121 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
      181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt
      721 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      781 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      841 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      901 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      961 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     1021 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     1081 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     1141 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     1201 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
     1261 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc caccatgact
     1321 gccgtccacg caggcaacat aaacttcaaa tgggacccca aaagtctgga gattaggact
     1381 ctggctgttg aaagactgtt ggagcctctt gttacacagg ttacaaccct ggtaaacacc
     1441 aatagtaaag ggccgtctaa taaaaagaga ggccgttcta agaaggccca tgttttggct
     1501 gcatctgttg aacaagcaac tgagaatttc ttggaaaagg gggataaaat tgcaaaagag
     1561 agccagtttc tcaaggagga gcttgtggtt gctgtagaag atgttcgaaa acaaggagat
     1621 ttgatgaaga gcgctgctgg ggagtttgca gatgatccat gctcttctgt gaagcgaggc
     1681 aacatggtcc gggcagctcg agctttgctc tctgccgtca cccggctgct gattctggct
     1741 gacatggcag atgtctacaa attacttgtt cagctgaaag ttgtggaaga tggtatattg
     1801 aaactgagga atgctggcaa tgaacaagac ttagggatac agtacaaagc tctgaaacca
     1861 gaagtagata agctgaacat catggcagca aaaagacaac aggaactaaa agatgtgggc
     1921 aatcgtgatc agatggctgc agctagagga atcctgcaga aaaatgttcc aatcctctat
     1981 actgcatccc aggcatgtct acaacaccct gatgttgcag cctataaggc caaccgggac
     2041 ttgatataca agcagctgca gcaggccgtc acaggaattt ctaatgcagc ccaggctact
     2101 gcatcagatg atgctgccca gcaccagggt ggcagtggcg gagagctggc atacgctctc
     2161 aacaactttg ataaacaaat cattgtggac cccttgagct tcagcgagga gcgctttagg
     2221 ccgtccctgg aggagcgcct ggaaagcatt attagtgggg ctgccctgat ggctgactca
     2281 tcctgcacac gtgatgaccg tcgggagcgc attgtggcag agtgtaatgc tgtccgccag
     2341 gctctacagg acctgctctc agaatacatg ggcaacgctg gacgtaaaga aagaagtgat
     2401 gcactcaact ctgcaataga taaaatgacc aagaagacca gggacttgcg tagacagctt
     2461 cgcaaagctg tcatggacca cgtatcagat tctttcctgg agaccaatgt ccctctatta
     2521 gtactgattg aagctgcaaa gaatggaaat gagaaagaag ttaaggaata tgcccaagtt
     2581 tttcgtgaac atgccaacaa actgattgag gttgccaacc tggcctgttc aatctccaac
     2641 aatgaagaag gcgtgaagct tgtccgaatg tctgcaagcc agttagaagc gctgtgtcct
     2701 caggttatca atgctgcatt ggcgttggca gcaaagccgc agagtaaact ggcccaagag
     2761 aacatggatc tttttaaaga gcaatgggaa aagcaagtcc gtgttctcac agatgctgtt
     2821 gatgacatta cttccatcga tgacttcttg gctgtctcag agaaccacat tttagaagat
     2881 gtgaacaagt gtgttattgc tctccaagag aaagacgtgg atgggctgga tcgcactgcc
     2941 ggtgccatcc gaggccgggc agcccgagtc attcatgtag tcacctcaga gatggacaac
     3001 tacgagccag gagtttacac agagaaggtt ctggaagcca ccaagctcct ctccaacaca
     3061 gtcatgccgc gcttcactga gcaggtggaa gcagctgtgg aagccctcag ctcggacccg
     3121 gcccagccca tggatgagaa tgagttcatc gacgcctccc gtctggtgta cgacggcatc
     3181 cgggacatcc ggaaagcagt gctgatgatc aggacccccg aggagttgga cgactctgac
     3241 ttcgagactg aagactttga tgtcagaagc aggaccagtg tccagacaga agatgaccag
     3301 ctgatagctg gccagagcgc ccgggcgatc atggctcagc ttcctcagga gcaaaaagca
     3361 aagattgcag aacaggtggc cagtttccag gaagagaaga gcaagctgga tgctgaagtg
     3421 tccaagtggg atgacagtgg caatgacatc attgtgctgg ccaagcagat gtgcatgatc
     3481 atgatggaga tgaccgactt cacccgaggc aaaggtccac tcaaaaacac ctctgacgtg
     3541 atcagtgctg ccaagaaaat tgcggaggcg ggatccagga tggacaagct cggccggacc
     3601 atcgcagacc attgcccaga ctcggcatgc aagcaggacc tgctggccta ccttcagcgc
     3661 atcgctctct actgccacca gctcaacatc tgcagcaaag tcaaggctga agtacagaac
     3721 cttggtggag agctggtcgt ctctggggtg gacagcgcca tgtccctgat ccaggcggcc
     3781 aagaacttga tgaatgctgt tgtgcagaca gtgaaggcgt cctacgtggc ttccaccaaa
     3841 taccagaagt cacaggggat ggcttccttg aaccttcctg ctgtgtcatg gaagatgaag
     3901 gcccctgaga agaagccgtt ggtgaagaga gagaagcagg atgagacgca gaccaagatt
     3961 aaacgagctt ctcagaagaa acacgtgaac ccagtgcagg ccctgagcga gttcaaagcc
     4021 atggacagca tcgccaccat ggtgagcaag ggcgaggaga ccacaatggg cgtaatcaag
     4081 cccgacatga agatcaagct gaagatggag ggcaacgtga atggccacgc cttcgtgatc
     4141 gagggcgagg gcgagggcaa gccctacgac ggcaccaaca ccatcaacct ggaggtgaag
     4201 gagggagccc ccctgccctt ctcctacgac attctgacca ccgcgttcgc ctacggcaac
     4261 agggccttca ccaagtaccc cgacgacatc cccaactact tcaagcagtc cttccccgag
     4321 ggctactctt gggagcgcac catgaccttc gaggacaagg gcatcgtgaa ggtgaagtcc
     4381 gacatctcca tggaggagga ctccttcatc tacgagatac acctcaaggg cgagaacttc
     4441 ccccccaacg gccccgtgat gcagaagaag accaccggct gggacgcctc caccgagagg
     4501 atgtacgtgc gcgacggcgt gctgaagggc gacgtcaagc acaagctgct gctggagggc
     4561 ggcggccacc accgcgttga cttcaagacc atctacaggg ccaagaaggc ggtgaagctg
     4621 cccgactatc actttgtgga ccaccgcatc gagatcctga accacgacaa ggactacaac
     4681 aaggtgaccg tttacgagag cgccgtggcc cgcaactcca ccgacggcat ggacgagctg
     4741 tacaaggggc caggtggtgc agggccaggt ggtgcagggc caggtggtgc agggccaggt
     4801 ggtgcagggc ccggtggtgc aggtccaggt ggtgcaggtc caggtggtgc aggtccaggt
     4861 ggtgctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4921 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     4981 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
     5041 cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac
     5101 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     5161 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     5221 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     5281 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     5341 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     5401 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     5461 aaccactacc tgagctacca gtccaaactg agcaaagacc ccaacgagaa gcgcgatcac
     5521 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     5581 aagtaggcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     5641 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     5701 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5761 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5821 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     5881 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     5941 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     6001 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     6061 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     6121 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     6181 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     6241 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     6301 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6361 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6421 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     6481 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     6541 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     6601 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     6661 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     6721 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     6781 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     6841 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     6901 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     6961 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     7021 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     7081 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     7141 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     7201 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     7261 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     7321 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     7381 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     7441 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     7501 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     7561 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     7621 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     7681 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     7741 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     7801 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     7861 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     7921 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     7981 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     8041 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     8101 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     8161 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaa
//
