LOCUS       sequence-458277-        7411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of GFP-alpha-catenin-delta-beta-H in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7411)
  AUTHORS   Noordstra I, Hermoso MD, Schimmel L, Bonfim-Melo A, Currin-Ross D, 
            Duong CN, Kalappurakkal JM, Morris RG, Vestweber D, Mayor S, Gordon 
            E, Roca-Cusachs P, Yap AS
  TITLE     An E-cadherin-actin clutch translates the mechanical force of 
            cortical flow for cell-cell contact to inhibit epithelial cell 
            locomotion.
  JOURNAL   Dev Cell. 2023 Sep 25;58(18):1748-1763.e6. doi: 
            10.1016/j.devcel.2023.06.011. Epub 2023 Jul 21.
  PUBMED    37480844
REFERENCE   2  (bases 1 to 7411)
  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.devcel.2023.06.011"; journalName: "Dev Cell"; date: 
            "2023-09-25- 25"; volume: "58"; issue: "18"; pages: "1748-1763.e6"
COMMENT     Sequence Label: GFP-alpha-catenin-Delta-beta-H
FEATURES             Location/Qualifiers
     source          1..7411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4199..4320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4236..4255)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4290..4309
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4327..4782)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4464..4485)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4676..4695
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4809..4913
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4915..5272
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4915..5106
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4938..4958)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5123..5258
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5307..6101
                     /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             5307..6101
                     /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(5361..5380)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5971..5990
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6289..6308)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6333..6380
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6709..7297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7198..7217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct catatgactg ctgtccatgc aggcaacata
     1381 aacttcaagt gggatcctaa aagtctagag atcaggactc tggcagttga gagactgttg
     1441 gagcctcttg ttacacaggt tacaaccctt gtaaacacca atagtaaagg gccctctaat
     1501 aagaagagag gtcgttctaa gaaggcccat gttttggctg catctgttga acaagcaact
     1561 gagaatttct tggagaaggg ggataaaatt gcgaaggaga gccagtttct caaggaggag
     1621 cttgtggctg ctgtagaaga tgttcgaaaa caaggtgatt tgatgaaggc tgctgcagga
     1681 gagttcgcag atgatccctg ctcttctgtg aagcgaggca acatggttcg ggcagctcga
     1741 gctttgctct ctgctgttac ccggttgctg attttggctg acatggcaga tgtctacaaa
     1801 ttacttgttc agctgaaagt tgtggaagat ggtatcttga agttgaggaa tgctggcaat
     1861 gaacaagact taggaatcca gtataaagcc ctaaaacctg aagtggataa gctgaacatt
     1921 atggcagcca aaagacaaca ggaattgaaa gatgttggcc atcgtgatca gatggctgca
     1981 gctagaggaa tcctgcagaa gaacgttccg atcctctata ctgcatccca ggcatgccta
     2041 cagcaccctg atgtcgcagc ctataaggcc aacagggacc tgatatacaa gcagctgcag
     2101 caggcggtca caggcatttc caatgcagcc caggccactg cctcagacga tgcctcacag
     2161 caccagggtg gaggaggagg agaactggca tatgcactca ataactttga caaacaaatc
     2221 attgtggacc ccttgagctt cagcgaggag cgctttaggc cttccctgga ggagcgtctg
     2281 gaaagcatca ttagtggggc tgccttgatg gccgactcgt cctgcacgcg tgatgaccgt
     2341 cgtgagcgaa ttgtggcaga gtgtaatgct gtccgccagg ccctgcagga cctgctttcg
     2401 gagtacatgg gcaatgctgg acgtaaagaa agaagtgatg cactcaattc tgcaatagat
     2461 aaaatgacca agaagaccag ggacttgcgt agacagctcc gcaaagctgt catggaccac
     2521 gtttcagatt ctttcctgga aaccaatgtt ccacttttgg tattgattga agctgcaaag
     2581 aatggaaatg agaaagaagt taaggagtat gcccaagttt tccgtgaaca tgccaacaaa
     2641 ttgattgagg ttgccaactt ggcctgttcc atctcaaata atgaagaagg tgtaaagctt
     2701 gttcgaatgt ctgcaagcca gttagaagcc ctctgtcctc aggttattaa tgctgcactg
     2761 gctttagcag caaaaccaca gagtaaactg gcccaagaga acatggatct ttttaaagaa
     2821 caatgggaaa aacaagtccg tgttctcaca gatgctgtcg atgacattac ttccattgat
     2881 gacttcttgg ctgtctcaga gaatcacatt ttggaagatg tgaacaaatg tgtcattgct
     2941 ctccaagaga aggatgtgga tggcctggac cgcacagctg gtgcaattcg aggccgggca
     3001 gcccgggtca ttcacgtagt cacctcagag atggacaact atgagccagg agtctacaca
     3061 gagaaggttc tggaagccac taagctgctc tccaacacag tcatgccacg ttttactgag
     3121 caagtagaag cagccgtgga agccctcagc tcggaccctg cccagcccat ggatgagaat
     3181 gagtttatcg atgcttcccg cctggtatat gatggcatcc gggacatcag gaaagcagtg
     3241 ctgatgataa ggacccctga ggagttggat gactctgact ttgagacaga agattttgat
     3301 gtcagaagca ggacgagcgt ccagacagaa gacgatcagc tgatagctgg ccagagtgcc
     3361 cgggcgatca tggctcagct tccccaggag caaaaagcga agattgcgga acaggtggcc
     3421 agcttccagg aagaaaagag caagctggat gctgaagtgt ccaaatggga cgacagtggc
     3481 aatgacatca ttgtgctggc caagcagatg tgcatgatta tgatggagat gacagacttt
     3541 acccgaggta aaggaccact caaaaataca tcggatgtca tcagtgctgc caagaaaatt
     3601 gctgaggcag gatccaggat ggacaagctt ggccgcacca ttgcagacca ttgccccgac
     3661 tcggcttgca agcaggacct gctggcctac ctgcaacgca tcgccctcta ctgccaccag
     3721 ctgaacatct gcagcaaggt caaggccgaa ttcggggtgg acagcgccat gtccctgatc
     3781 caggcagcca agaacttgat gaatgctgtg gtgcagacag tgaaggcatc ctacgtcgcc
     3841 tctaccaaat accaaaagtc acagggtatg gcttccctca accttcctgc tgtgtcatgg
     3901 aagatgaagg caccagagaa aaagccattg gtgaagagag agaaacagga tgagacacag
     3961 accaagatta aacgggcatc tcagaagaag cacgtgaacc cggtgcaggc cctcagcgag
     4021 ttcaaagcta tggacagcat ctaatgcagt cgacggtacc gcgggcccgg gatccaccgg
     4081 atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4141 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4201 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4261 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4321 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4381 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4441 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4501 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4561 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4621 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4681 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4741 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4801 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4861 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4921 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4981 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     5041 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     5101 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5161 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5221 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     5281 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5341 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5401 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5461 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     5521 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5581 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5641 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5701 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5761 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5821 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5881 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5941 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6001 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6061 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     6121 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6181 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6241 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     6301 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     6361 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     6421 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     6481 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     6541 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6601 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6661 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6721 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6781 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6841 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6901 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6961 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7021 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7081 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7141 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7201 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7261 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7321 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7381 ttctgtggat aaccgtatta ccgccatgca t
//
