LOCUS       sequence-195021-        8613 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of E-cadherin-GFP-N-WASP delta VCA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8613)
  AUTHORS   Wu SK, Gomez GA, Michael M, Verma S, Cox HL, Lefevre JG, Parton RG, 
            Hamilton NA, Neufeld Z, Yap AS
  TITLE     Cortical F-actin stabilization generates apical-lateral patterns of 
            junctional contractility that integrate cells into epithelia.
  JOURNAL   Nat Cell Biol. 2014 Feb;16(2):167-78. doi: 10.1038/ncb2900. Epub 
            2014 Jan 12.
  PUBMED    24413434
REFERENCE   2  (bases 1 to 8613)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1038/ncb2900"; 
            journalName: "Nat Cell Biol"; date: "2014-02"; volume: "16"; issue: 
            "2"; pages: "167-78"
COMMENT     Sequence Label: pE-cadherin-GFP-N-WASP deltaVCA
FEATURES             Location/Qualifiers
     source          1..8613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             50..844
                     /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             50..844
                     /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(104..123)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     714..733
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1032..1051)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1076..1123
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1452..2040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1941..1960
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2215..2518
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2519..2722
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2673..2693
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    2745..2825
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      5487..5496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5493..5495,5496..5498,5499..6209)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5493..5495,5496..5498,5499..6209)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5538..5559)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5799..5818)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6146..6167
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    7555..7676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7592..7611)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7646..7665
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(7683..8138)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7820..7841)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8032..8051
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        8165..8269
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(8271..8613,1..15)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        8271..8462
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(8294..8314)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(8479..8613,1)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
       61 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
      121 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
      181 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
      241 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
      301 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
      361 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
      421 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
      481 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
      541 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
      601 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
      661 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
      721 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
      781 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
      841 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
      901 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
      961 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     1021 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     1081 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     1141 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     1201 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     1261 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     1321 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     1381 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1441 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1501 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1561 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     1621 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1681 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1741 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1801 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1861 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1921 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1981 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2041 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2101 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     2161 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     2221 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     2281 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     2341 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     2401 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     2461 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     2521 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     2581 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     2641 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     2701 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
     2761 tcagatctcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca
     2821 ccggtatggg agcccggtgc cgcagctttt ccgcgctcct gctcctgctg caggtctcct
     2881 catggctttg ccaggagctg gagcctgagt cctgcagtcc cggcttcagt tccgaggtct
     2941 acaccttccc ggtgccggag aggcacctgg agagaggcca tgtcctgggc agagtgagat
     3001 ttgaaggatg caccggccgg ccaaggacag ccttcttttc ggaagactcc cgattcaaag
     3061 tggcgacaga cggcaccatc acagtgaagc ggcatctaaa gctccacaag ctggagacca
     3121 gtttcctcgt ccgcgcccgg gactccagtc atagggagct gtctaccaaa gtgacgctga
     3181 agtccatggg gcaccaccat caccggcacc accaccgcga ccctgcctct gaatccaacc
     3241 cagagctgct catgtttccc agcgtgtacc caggtctcag aagacagaaa cgagactggg
     3301 tcatccctcc catcagctgc cccgaaaatg aaaagggtga attcccaaag aacctggttc
     3361 agatcaaatc caacagggac aaagaaacaa aggttttcta cagcatcacc ggccaaggag
     3421 ctgacaaacc ccccgttggc gttttcatca ttgagaggga gacaggctgg ctgaaagtga
     3481 cacagcctct ggatagagaa gccattgcca agtacatcct ctattctcat gccgtgtcat
     3541 caaatgggga agcggtggag gatcccatgg agatagtgat cacagtgaca gatcagaatg
     3601 acaacaggcc agagtttacc caggaggtgt ttgagggatc cgttgcagaa ggcgctgttc
     3661 caggaacctc cgtgatgaag gtctcagcca ccgatgcaga cgatgacgtc aacacctaca
     3721 acgctgccat cgcctacacc atcgtcagcc aggatcctga gctgcctcac aaaaacatgt
     3781 tcactgtcaa tagggacacc ggggtcatca gtgtgctcac ctctgggctg gaccgagaga
     3841 gttaccctac atacactctg gtggttcagg ctgctgacct tcaaggcgaa ggcttgagca
     3901 caacagccaa ggctgtgatc actgtcaagg atattaatga caacgctcct gtcttcaacc
     3961 cgagcacgta tcagggtcaa gtgcctgaga atgaggtcaa tgcccggatc gccacactca
     4021 aagtgaccga tgatgatgcc cccaacactc cggcgtggaa agctgtgtac accgtagtca
     4081 acgatcctga ccagcagttc gttgtcgtca cagaccccac gaccaatgat ggcattttga
     4141 aaacagccaa gggcttggat tttgaggcca agcagcaata catccttcat gtgagagtgg
     4201 agaacgagga accctttgag gggtctcttg tcccttccac agccactgtc actgtggacg
     4261 tggtagacgt gaatgaagcc cccatcttta tgcctgcgga gaggagagtc gaagtgcccg
     4321 aagactttgg tgtgggtcag gaaatcacat cttataccgc tcgagagccg gacacgttca
     4381 tggatcagaa gatcacgtat cggatttgga gggacactgc caactggctg gagattaacc
     4441 cagagactgg tgccattttc acgcgcgctg agatggacag agaagacgct gagcatgtga
     4501 agaacagcac atatgtagct ctcatcatcg ccacagatga tggttcaccc attgccactg
     4561 gcacgggcac tcttctcctg gtcctgttag acgtcaatga caacgctccc atcccagaac
     4621 ctcgaaacat gcagttctgc cagaggaacc cacagcctca tatcatcacc atcttggatc
     4681 cagaccttcc ccccaacacg tcccccttta ctgctgagct aacccatggg gccagcgtca
     4741 actggaccat tgagtataat gacgcagctc aagaatctct cattttgcaa ccaagaaagg
     4801 acttagagat tggcgaatac aaaatccatc tcaagctcgc ggataaccag aacaaagacc
     4861 aggtgaccac gttggacgtc catgtgtgtg actgtgaagg gacggtcaac aactgcatga
     4921 aggcgggaat cgtggcagca ggattgcaag ttcctgccat cctcggaatc cttggaggga
     4981 tcctcgccct gctgattctg atcctgctgc tcctactgtt tctacggagg agaacggtgg
     5041 tcaaagagcc cctgctgcca ccagatgatg atacccggga caatgtgtat tactatgatg
     5101 aagaaggagg tggagaagaa gaccaggact ttgatttgag ccagctgcac aggggcctgg
     5161 atgcccgacc ggaagtgact cgaaatgatg tggctcccac cctcatgagc gtgccccagt
     5221 atcgtccccg tcctgccaat cctgatgaaa ttggaaactt catcgatgaa aacctgaagg
     5281 cagccgacag cgaccccacg gcaccccctt acgactctct gttggtgttc gattacgagg
     5341 gcagtggttc tgaagccgct agcctgagct cactgaactc ctctgagtcg gatcaggacc
     5401 aggactacga ttatctgaac gagtggggca accgattcaa gaagctggcg gacatgtacg
     5461 gcggtggcga ggacgacaaa ccggtcgcca ccatggtgag caagggcgag gagctgttca
     5521 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     5581 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     5641 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     5701 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     5761 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     5821 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     5881 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     5941 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     6001 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     6061 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     6121 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     6181 tcactctcgg catggacgag ctgtacaagg ggtctacatc tggatctggg aagccgggtt
     6241 ctggtgaggg ttctatgagc tcgggccagc agcccccgcg gagggtcacc aacgtgggct
     6301 ccctgctgct caccccgcaa gagaacgagt ctctcttctc cttcctcggc aagaaatgtg
     6361 tgactatgtc ttcagcagtg gtgcagttgt atgcagcaga tcggaactgt atgtgggcaa
     6421 agaagtgcag tggtgtcgct tgtctggtta aggacaatcc tcagagatct tattttttaa
     6481 gaatatttga cattaaggat gggaaattac tgtgggaaca agagctatac aataactttg
     6541 tatataatag tcctagagga tattttcata cctttgctgg agatacttgt caagtagctc
     6601 ttaattttgc caatgaagaa gaagcaaaaa agttccgaaa agcagttaca gacctgttgg
     6661 gccgacgaca aaggaaatct gaaaaaagac gagatgctcc aaatggtccc aatctaccca
     6721 tggctacagt tgacataaaa aatccagaaa tcacaacaaa tagattttat ggttcacaag
     6781 tcaacaacat ctcccacacc aaagaaaaga agaaaggaaa agctaaaaag aagagattaa
     6841 ccaaggcaga tattggaaca ccaagtaatt tccagcacat tgggcatgtt ggatgggatc
     6901 caaatacagg ttttgatcta aataatttgg atccagaatt aaagaatctt tttgatatgt
     6961 gtgggatctc tgaggcccag cttaaagaca gagaaacatc aaaagttatt tatgacttta
     7021 ttgaaaaaac aggaggtgta gaagctgtta aaaacgaact ccgaagacaa gcaccaccac
     7081 cacctccacc ctcgagagga ggacctcccc ctcctcctcc ccctcctcat agctcaggtc
     7141 ctcctccccc tcctgcccga ggaagggggg ctcctcctcc cccaccttca agagctccta
     7201 ctgctgcacc tccacctcca cctccttcta ggcctggtgt tgttgttcct ccgccccctc
     7261 caaacaggat gtaccctcct ccaccaccag cgctgccctc ttcagcacct tcaggccctc
     7321 caccacctcc acctccatct atggcagggt ccacagcacc accacctcct cctccacctc
     7381 cgcccccacc agggccacca cctccccctg gcctgccttc tgatggtgac tgagcggccg
     7441 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     7501 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     7561 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     7621 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     7681 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     7741 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     7801 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     7861 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     7921 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     7981 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     8041 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     8101 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     8161 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     8221 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     8281 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     8341 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     8401 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     8461 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     8521 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     8581 ttccagaagt agtgaggagg cttttttgga ggc
//
