LOCUS       sequence-458273-        7441 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Transient expression of GFP-alpha-cateninWT in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7441)
  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 7441)
  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: 
            "10.1016/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-cateninWT
FEATURES             Location/Qualifiers
     source          1..7441
                     /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    4229..4350
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4266..4285)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4320..4339
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4357..4812)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4494..4515)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4706..4725
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4839..4943
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4945..5302
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4945..5136
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4968..4988)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5153..5288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5337..6131
                     /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             5337..6131
                     /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(5391..5410)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6001..6020
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6319..6338)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6363..6410
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6739..7327
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7228..7247
                     /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 caaggccgag gtgcagaatc tcggcgggga gcttgttgtc
     3781 tctggggtgg acagcgccat gtccctgatc caggcagcca agaacttgat gaatgctgtg
     3841 gtgcagacag tgaaggcatc ctacgtcgcc tctaccaaat accaaaagtc acagggtatg
     3901 gcttccctca accttcctgc tgtgtcatgg aagatgaagg caccagagaa aaagccattg
     3961 gtgaagagag agaaacagga tgagacacag accaagatta aacgggcatc tcagaagaag
     4021 cacgtgaacc cggtgcaggc cctcagcgag ttcaaagcta tggacagcat ctaatgcagt
     4081 cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     4141 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     4201 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     4261 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4321 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     4381 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4441 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4501 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4561 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4621 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4681 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4741 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4801 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4861 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4921 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4981 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5041 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5101 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5161 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5221 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5281 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     5341 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5401 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5461 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     5521 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5581 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5641 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5701 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5761 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5821 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     5881 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5941 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6001 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6061 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6121 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6181 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6241 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6301 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     6361 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     6421 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     6481 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     6541 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     6601 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6661 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6721 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6781 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6841 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6901 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6961 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7021 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7081 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7141 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7201 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7261 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7321 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7381 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     7441 t
//
