LOCUS       sequence-246103-        7734 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. See Depositor 
            Comments for plasmid construction..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7734)
  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 7734)
  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 DeltaTS (delta570 to 670 aa in aCat TS WT)
FEATURES             Location/Qualifiers
     source          1..7734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        777..1080
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1081..1284
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1235..1255
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2933..3640
                     /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(3761..3763,3764..3766,3767..4477)
                     /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(3806..3827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4067..4086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4414..4435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5238..5359
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5275..5294)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5329..5348
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5366..5821)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5503..5524)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5715..5734
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5848..5952
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5954..6311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5954..6145
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5977..5997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6162..6297
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6346..7140
                     /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             6346..7140
                     /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(6400..6419)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7010..7029
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7328..7347)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7372..7419
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
      721 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      781 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      841 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      901 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      961 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1021 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1081 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     1141 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     1201 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     1261 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcatgactgc
     1321 cgtccacgca ggcaacataa acttcaaatg ggaccccaaa agtctggaga ttaggactct
     1381 ggctgttgaa agactgttgg agcctcttgt tacacaggtt acaaccctgg taaacaccaa
     1441 tagtaaaggg ccgtctaata aaaagagagg ccgttctaag aaggcccatg ttttggctgc
     1501 atctgttgaa caagcaactg agaatttctt ggaaaagggg gataaaattg caaaagagag
     1561 ccagtttctc aaggaggagc ttgtggttgc tgtagaagat gttcgaaaac aaggagattt
     1621 gatgaagagc gctgctgggg agtttgcaga tgatccatgc tcttctgtga agcgaggcaa
     1681 catggtccgg gcagctcgag ctttgctctc tgccgtcacc cggctgctga ttctggctga
     1741 catggcagat gtctacaaat tacttgttca gctgaaagtt gtggaagatg gtatattgaa
     1801 actgaggaat gctggcaatg aacaagactt agggatacag tacaaagctc tgaaaccaga
     1861 agtagataag ctgaacatca tggcagcaaa aagacaacag gaactaaaag atgtgggcaa
     1921 tcgtgatcag atggctgcag ctagaggaat cctgcagaaa aatgttccaa tcctctatac
     1981 tgcatcccag gcatgtctac aacaccctga tgttgcagcc tataaggcca accgggactt
     2041 gatatacaag cagctgcagc aggccgtcac aggaatttct aatgcagccc aggctactgc
     2101 atcagatgat gctgcccagc accagggtgg cagtggcgga gagctggcat acgctctcaa
     2161 caactttgat aaacaaatca ttgtggaccc cttgagcttc agcgaggagc gctttaggcc
     2221 gtccctggag gagcgcctgg aaagcattat tagtggggct gccctgatgg ctgactcatc
     2281 ctgcacacgt gatgaccgtc gggagcgcat tgtggcagag tgtaatgctg tccgccaggc
     2341 tctacaggac ctgctctcag aatacatggg caacgctgga cgtaaagaaa gaagtgatgc
     2401 actcaactct gcaatagata aaatgaccaa gaagaccagg gacttgcgta gacagcttcg
     2461 caaagctgtc atggaccacg tatcagattc tttcctggag accaatgtcc ctctattagt
     2521 actgattgaa gctgcaaaga atggaaatga gaaagaagtt aaggaatatg cccaagtttt
     2581 tcgtgaacat gccaacaaac tgattgaggt tgccaacctg gcctgttcaa tctccaacaa
     2641 tgaagaaggc gtgaagcttg tccgaatgtc tgcaagccag ttagaagcgc tgtgtcctca
     2701 ggttatcaat gctgcattgg cgttggcagc aaagccgcag agtaaactgg cccaagagaa
     2761 catggatctt tttaaagagc aatgggaaaa gcaagtccgt gttctcacag atgctgttga
     2821 tgacattact tccatcgatt cccgggcgat catggctcag cttcctcagg agcaaaaagc
     2881 aaagattgca gaacaggtgg ccagtttcca ggaagagaag agcaagctcg agatggtgag
     2941 caagggcgag gagaccacaa tgggcgtaat caagcccgac atgaagatca agctgaagat
     3001 ggagggcaac gtgaatggcc acgccttcgt gatcgagggc gagggcgagg gcaagcccta
     3061 cgacggcacc aacaccatca acctggaggt gaaggaggga gcccccctgc ccttctccta
     3121 cgacattctg accaccgcgt tcgcctacgg caacagggcc ttcaccaagt accccgacga
     3181 catccccaac tacttcaagc agtccttccc cgagggctac tcttgggagc gcaccatgac
     3241 cttcgaggac aagggcatcg tgaaggtgaa gtccgacatc tccatggagg aggactcctt
     3301 catctacgag atacacctca agggcgagaa cttccccccc aacggccccg tgatgcagaa
     3361 gaagaccacc ggctgggacg cctccaccga gaggatgtac gtgcgcgacg gcgtgctgaa
     3421 gggcgacgtc aagcacaagc tgctgctgga gggcggcggc caccaccgcg ttgacttcaa
     3481 gaccatctac agggccaaga aggcggtgaa gctgcccgac tatcactttg tggaccaccg
     3541 catcgagatc ctgaaccacg acaaggacta caacaaggtg accgtttacg agagcgccgt
     3601 ggcccgcaac tccaccgacg gcatggacga gctgtacaag gggccaggtg gtgcagggcc
     3661 aggtggtgca gggccaggtg gtgcagggcc aggtggtgca gggcccggtg gtgcaggtcc
     3721 aggtggtgca ggtccaggtg gtgcaggtcc aggtggtgct atggtgagca agggcgagga
     3781 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3841 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct
     3901 gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgggcta
     3961 cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4021 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     4081 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4141 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     4201 cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     4261 gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac
     4321 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     4381 actgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     4441 cgccgggatc actctcggca tggacgagct gtacaaggcg gccgcgctgg atgctgaagt
     4501 gtccaagtgg gatgacagtg gcaatgacat cattgtgctg gccaagcaga tgtgcatgat
     4561 catgatggag atgaccgact tcacccgagg caaaggtcca ctcaaaaaca cctctgacgt
     4621 gatcagtgct gccaagaaaa ttgcggaggc gggatccagg atggacaagc tcggccggac
     4681 catcgcagac cattgcccag actcggcatg caagcaggac ctgctggcct accttcagcg
     4741 catcgctctc tactgccacc agctcaacat ctgcagcaaa gtcaaggctg aagtacagaa
     4801 ccttggtgga gagctggtcg tctctggggt ggacagcgcc atgtccctga tccaggcggc
     4861 caagaacttg atgaatgctg ttgtgcagac agtgaaggcg tcctacgtgg cttccaccaa
     4921 ataccagaag tcacagggga tggcttcctt gaaccttcct gctgtgtcat ggaagatgaa
     4981 ggcccctgag aagaagccgt tggtgaagag agagaagcag gatgagacgc agaccaagat
     5041 taaacgagct tctcagaaga aacacgtgaa cccagtgcag gccctgagcg agttcaaagc
     5101 catggacagc atctgagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     5161 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     5221 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5281 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5341 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     5401 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5461 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5521 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5581 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5641 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5701 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5761 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5821 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5881 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5941 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     6001 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6061 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6121 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6181 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6241 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6301 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     6361 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6421 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6481 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     6541 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6601 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6661 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6721 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6781 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6841 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     6901 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6961 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     7021 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     7081 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     7141 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     7201 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     7261 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     7321 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     7381 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     7441 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     7501 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     7561 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     7621 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7681 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aaga
//
