LOCUS       sequence-246086-        7604 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 7604)
  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 7604)
  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 TL2 (1-697 mTFP1-TSmod-Venus STOP)
FEATURES             Location/Qualifiers
     source          1..7604
                     /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"
     CDS             3435..4142
                     /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(4263..4265,4266..4268,4269..4982)
                     /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(4308..4329)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4569..4588)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4916..4937
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5104..5225
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5141..5160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5195..5214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5232..5687)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5369..5390)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5581..5600
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5714..5818
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5820..6177
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5820..6011
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5843..5863)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6028..6163
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6212..7006
                     /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             6212..7006
                     /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(6266..6285)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6876..6895
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7194..7213)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7238..7285
                     /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 taccggactc
     1321 agatctcgag cgccaccatg actgccgtcc acgcaggcaa cataaacttc aaatgggacc
     1381 ccaaaagtct ggagattagg actctggctg ttgaaagact gttggagcct cttgttacac
     1441 aggttacaac cctggtaaac accaatagta aagggccgtc taataaaaag agaggccgtt
     1501 ctaagaaggc ccatgttttg gctgcatctg ttgaacaagc aactgagaat ttcttggaaa
     1561 agggggataa aattgcaaaa gagagccagt ttctcaagga ggagcttgtg gttgctgtag
     1621 aagatgttcg aaaacaagga gatttgatga agagcgctgc tggggagttt gcagatgatc
     1681 catgctcttc tgtgaagcga ggcaacatgg tccgggcagc tcgagctttg ctctctgccg
     1741 tcacccggct gctgattctg gctgacatgg cagatgtcta caaattactt gttcagctga
     1801 aagttgtgga agatggtata ttgaaactga ggaatgctgg caatgaacaa gacttaggga
     1861 tacagtacaa agctctgaaa ccagaagtag ataagctgaa catcatggca gcaaaaagac
     1921 aacaggaact aaaagatgtg ggcaatcgtg atcagatggc tgcagctaga ggaatcctgc
     1981 agaaaaatgt tccaatcctc tatactgcat cccaggcatg tctacaacac cctgatgttg
     2041 cagcctataa ggccaaccgg gacttgatat acaagcagct gcagcaggcc gtcacaggaa
     2101 tttctaatgc agcccaggct actgcatcag atgatgctgc ccagcaccag ggtggcagtg
     2161 gcggagagct ggcatacgct ctcaacaact ttgataaaca aatcattgtg gaccccttga
     2221 gcttcagcga ggagcgcttt aggccgtccc tggaggagcg cctggaaagc attattagtg
     2281 gggctgccct gatggctgac tcatcctgca cacgtgatga ccgtcgggag cgcattgtgg
     2341 cagagtgtaa tgctgtccgc caggctctac aggacctgct ctcagaatac atgggcaacg
     2401 ctggacgtaa agaaagaagt gatgcactca actctgcaat agataaaatg accaagaaga
     2461 ccagggactt gcgtagacag cttcgcaaag ctgtcatgga ccacgtatca gattctttcc
     2521 tggagaccaa tgtccctcta ttagtactga ttgaagctgc aaagaatgga aatgagaaag
     2581 aagttaagga atatgcccaa gtttttcgtg aacatgccaa caaactgatt gaggttgcca
     2641 acctggcctg ttcaatctcc aacaatgaag aaggcgtgaa gcttgtccga atgtctgcaa
     2701 gccagttaga agcgctgtgt cctcaggtta tcaatgctgc attggcgttg gcagcaaagc
     2761 cgcagagtaa actggcccaa gagaacatgg atctttttaa agagcaatgg gaaaagcaag
     2821 tccgtgttct cacagatgct gttgatgaca ttacttccat cgatgacttc ttggctgtct
     2881 cagagaacca cattttagaa gatgtgaaca agtgtgttat tgctctccaa gagaaagacg
     2941 tggatgggct ggatcgcact gccggtgcca tccgaggccg ggcagcccga gtcattcatg
     3001 tagtcacctc agagatggac aactacgagc caggagttta cacagagaag gttctggaag
     3061 ccaccaagct cctctccaac acagtcatgc cgcgcttcac tgagcaggtg gaagcagctg
     3121 tggaagccct cagctcggac ccggcccagc ccatggatga gaatgagttc atcgacgcct
     3181 cccgtctggt gtacgacggc atccgggaca tccggaaagc agtgctgatg atcaggaccc
     3241 ccgaggagtt ggacgactct gacttcgaga ctgaagactt tgatgtcaga agcaggacca
     3301 gtgtccagac agaagatgac cagctgatag ctggccagag cgcccgggcg atcatggctc
     3361 agcttcctca ggagcaaaaa gcaaagattg cagaacaggt ggccagtttc caggaagaga
     3421 agagcaagct ggagatggtg agcaagggcg aggagaccac aatgggcgta atcaagcccg
     3481 acatgaagat caagctgaag atggagggca acgtgaatgg ccacgccttc gtgatcgagg
     3541 gcgagggcga gggcaagccc tacgacggca ccaacaccat caacctggag gtgaaggagg
     3601 gagcccccct gcccttctcc tacgacattc tgaccaccgc gttcgcctac ggcaacaggg
     3661 ccttcaccaa gtaccccgac gacatcccca actacttcaa gcagtccttc cccgagggct
     3721 actcttggga gcgcaccatg accttcgagg acaagggcat cgtgaaggtg aagtccgaca
     3781 tctccatgga ggaggactcc ttcatctacg agatacacct caagggcgag aacttccccc
     3841 ccaacggccc cgtgatgcag aagaagacca ccggctggga cgcctccacc gagaggatgt
     3901 acgtgcgcga cggcgtgctg aagggcgacg tcaagcacaa gctgctgctg gagggcggcg
     3961 gccaccaccg cgttgacttc aagaccatct acagggccaa gaaggcggtg aagctgcccg
     4021 actatcactt tgtggaccac cgcatcgaga tcctgaacca cgacaaggac tacaacaagg
     4081 tgaccgttta cgagagcgcc gtggcccgca actccaccga cggcatggac gagctgtaca
     4141 aggggccagg tggtgcaggg ccaggtggtg cagggccagg tggtgcaggg ccaggtggtg
     4201 cagggcccgg tggtgcaggt ccaggtggtg caggtccagg tggtgcaggt ccaggtggtg
     4261 ctatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     4321 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     4381 acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca
     4441 ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
     4501 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     4561 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4621 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4681 acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga
     4741 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     4801 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     4861 actacctgag ctaccagtcc aaactgagca aagaccccaa cgagaagcgc gatcacatgg
     4921 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     4981 aggcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     5041 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     5101 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5161 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5221 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     5281 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     5341 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     5401 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     5461 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     5521 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     5581 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     5641 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     5701 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5761 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     5821 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     5881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     5941 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     6001 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     6061 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     6121 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     6181 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     6241 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     6301 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     6361 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     6421 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     6481 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     6541 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     6601 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     6661 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     6721 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     6781 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     6841 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6901 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6961 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     7021 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     7081 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     7141 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     7201 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     7261 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     7321 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     7381 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     7441 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     7501 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7561 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaa
//
