LOCUS       Exported                8240 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  FRET-based tension sensor (TS) technology to develop a probe for 
            molecular-level tension across alphaE-catenin..
ACCESSION   .
VERSION     .
KEYWORDS    aCat TS WT (1-697 mTFP1-TSmod-Venus 698-907)
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8240
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             3439..4146
                     /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             4267..4983
                     /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(4312..4333)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4573..4592)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4920..4941
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5744..5865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5781..5800)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5835..5854
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5872..6327)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6009..6030)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6221..6240
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6354..6458
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6460..6817
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(6483..6503)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6668..6803
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6730..6749
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6852..7646
                     /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(6906..6925)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7516..7535
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7834..7853)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7878..7925
                     /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 gcgctaccgg
     1321 actcagatct cgagcgccac catgactgcc gtccacgcag gcaacataaa cttcaaatgg
     1381 gaccccaaaa gtctggagat taggactctg gctgttgaaa gactgttgga gcctcttgtt
     1441 acacaggtta caaccctggt aaacaccaat agtaaagggc cgtctaataa aaagagaggc
     1501 cgttctaaga aggcccatgt tttggctgca tctgttgaac aagcaactga gaatttcttg
     1561 gaaaaggggg ataaaattgc aaaagagagc cagtttctca aggaggagct tgtggttgct
     1621 gtagaagatg ttcgaaaaca aggagatttg atgaagagcg ctgctgggga gtttgcagat
     1681 gatccatgct cttctgtgaa gcgaggcaac atggtccggg cagctcgagc tttgctctct
     1741 gccgtcaccc ggctgctgat tctggctgac atggcagatg tctacaaatt acttgttcag
     1801 ctgaaagttg tggaagatgg tatattgaaa ctgaggaatg ctggcaatga acaagactta
     1861 gggatacagt acaaagctct gaaaccagaa gtagataagc tgaacatcat ggcagcaaaa
     1921 agacaacagg aactaaaaga tgtgggcaat cgtgatcaga tggctgcagc tagaggaatc
     1981 ctgcagaaaa atgttccaat cctctatact gcatcccagg catgtctaca acaccctgat
     2041 gttgcagcct ataaggccaa ccgggacttg atatacaagc agctgcagca ggccgtcaca
     2101 ggaatttcta atgcagccca ggctactgca tcagatgatg ctgcccagca ccagggtggc
     2161 agtggcggag agctggcata cgctctcaac aactttgata aacaaatcat tgtggacccc
     2221 ttgagcttca gcgaggagcg ctttaggccg tccctggagg agcgcctgga aagcattatt
     2281 agtggggctg ccctgatggc tgactcatcc tgcacacgtg atgaccgtcg ggagcgcatt
     2341 gtggcagagt gtaatgctgt ccgccaggct ctacaggacc tgctctcaga atacatgggc
     2401 aacgctggac gtaaagaaag aagtgatgca ctcaactctg caatagataa aatgaccaag
     2461 aagaccaggg acttgcgtag acagcttcgc aaagctgtca tggaccacgt atcagattct
     2521 ttcctggaga ccaatgtccc tctattagta ctgattgaag ctgcaaagaa tggaaatgag
     2581 aaagaagtta aggaatatgc ccaagttttt cgtgaacatg ccaacaaact gattgaggtt
     2641 gccaacctgg cctgttcaat ctccaacaat gaagaaggcg tgaagcttgt ccgaatgtct
     2701 gcaagccagt tagaagcgct gtgtcctcag gttatcaatg ctgcattggc gttggcagca
     2761 aagccgcaga gtaaactggc ccaagagaac atggatcttt ttaaagagca atgggaaaag
     2821 caagtccgtg ttctcacaga tgctgttgat gacattactt ccatcgatga cttcttggct
     2881 gtctcagaga accacatttt agaagatgtg aacaagtgtg ttattgctct ccaagagaaa
     2941 gacgtggatg ggctggatcg cactgccggt gccatccgag gccgggcagc ccgagtcatt
     3001 catgtagtca cctcagagat ggacaactac gagccaggag tttacacaga gaaggttctg
     3061 gaagccacca agctcctctc caacacagtc atgccgcgct tcactgagca ggtggaagca
     3121 gctgtggaag ccctcagctc ggacccggcc cagcccatgg atgagaatga gttcatcgac
     3181 gcctcccgtc tggtgtacga cggcatccgg gacatccgga aagcagtgct gatgatcagg
     3241 acccccgagg agttggacga ctctgacttc gagactgaag actttgatgt cagaagcagg
     3301 accagtgtcc agacagaaga tgaccagctg atagctggcc agagcgcccg ggcgatcatg
     3361 gctcagcttc ctcaggagca aaaagcaaag attgcagaac aggtggccag tttccaggaa
     3421 gagaagagca agctcgagat ggtgagcaag ggcgaggaga ccacaatggg cgtaatcaag
     3481 cccgacatga agatcaagct gaagatggag ggcaacgtga atggccacgc cttcgtgatc
     3541 gagggcgagg gcgagggcaa gccctacgac ggcaccaaca ccatcaacct ggaggtgaag
     3601 gagggagccc ccctgccctt ctcctacgac attctgacca ccgcgttcgc ctacggcaac
     3661 agggccttca ccaagtaccc cgacgacatc cccaactact tcaagcagtc cttccccgag
     3721 ggctactctt gggagcgcac catgaccttc gaggacaagg gcatcgtgaa ggtgaagtcc
     3781 gacatctcca tggaggagga ctccttcatc tacgagatac acctcaaggg cgagaacttc
     3841 ccccccaacg gccccgtgat gcagaagaag accaccggct gggacgcctc caccgagagg
     3901 atgtacgtgc gcgacggcgt gctgaagggc gacgtcaagc acaagctgct gctggagggc
     3961 ggcggccacc accgcgttga cttcaagacc atctacaggg ccaagaaggc ggtgaagctg
     4021 cccgactatc actttgtgga ccaccgcatc gagatcctga accacgacaa ggactacaac
     4081 aaggtgaccg tttacgagag cgccgtggcc cgcaactcca ccgacggcat ggacgagctg
     4141 tacaaggggc caggtggtgc agggccaggt ggtgcagggc caggtggtgc agggccaggt
     4201 ggtgcagggc ccggtggtgc aggtccaggt ggtgcaggtc caggtggtgc aggtccaggt
     4261 ggtgctatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     4321 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     4381 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
     4441 cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac
     4501 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     4561 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     4621 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     4681 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
     4741 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
     4801 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     4861 aaccactacc tgagctacca gtccaaactg agcaaagacc ccaacgagaa gcgcgatcac
     4921 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     4981 aaggcggccg cgctggatgc tgaagtgtcc aagtgggatg acagtggcaa tgacatcatt
     5041 gtgctggcca agcagatgtg catgatcatg atggagatga ccgacttcac ccgaggcaaa
     5101 ggtccactca aaaacacctc tgacgtgatc agtgctgcca agaaaattgc ggaggcggga
     5161 tccaggatgg acaagctcgg ccggaccatc gcagaccatt gcccagactc ggcatgcaag
     5221 caggacctgc tggcctacct tcagcgcatc gctctctact gccaccagct caacatctgc
     5281 agcaaagtca aggctgaagt acagaacctt ggtggagagc tggtcgtctc tggggtggac
     5341 agcgccatgt ccctgatcca ggcggccaag aacttgatga atgctgttgt gcagacagtg
     5401 aaggcgtcct acgtggcttc caccaaatac cagaagtcac aggggatggc ttccttgaac
     5461 cttcctgctg tgtcatggaa gatgaaggcc cctgagaaga agccgttggt gaagagagag
     5521 aagcaggatg agacgcagac caagattaaa cgagcttctc agaagaaaca cgtgaaccca
     5581 gtgcaggccc tgagcgagtt caaagccatg gacagcatct gagcggccgc gactctagat
     5641 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     5701 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     5761 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     5821 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     5881 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5941 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     6001 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     6061 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     6121 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     6181 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     6241 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     6301 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     6361 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6421 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     6481 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6541 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     6601 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     6661 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6721 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6781 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     6841 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     6901 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     6961 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     7021 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     7081 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     7141 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     7201 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     7261 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     7321 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     7381 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     7441 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     7501 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     7561 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     7621 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     7681 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     7741 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     7801 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     7861 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     7921 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     7981 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     8041 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     8101 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     8161 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8221 gtcagacccc gtagaaaaga
//
