LOCUS       sequence-244395-        7503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of E-cadherin-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7503)
  AUTHORS   Truffi M, Dubreuil V, Liang X, Vacaresse N, Nigon F, Han SP, Yap AS,
            Gomez GA, Sap J
  TITLE     RPTPalpha controls epithelial adherens junctions, linking E-cadherin
            engagement to c-Src-mediated phosphorylation of cortactin.
  JOURNAL   J Cell Sci. 2014 Jun 1;127(Pt 11):2420-32. doi: 10.1242/jcs.134379. 
            Epub 2014 Mar 20.
  PUBMED    24652832
REFERENCE   2  (bases 1 to 7503)
  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"; journalName: "J Cell 
            Sci."; date: "2014-06-1"; volume: "127(Pt 11):2420-32. doi"; pages: 
            " 10.1242/jcs.134379. Epub 2014 Mar 20"
COMMENT     Sequence Label: mouse E-cadherin GFP (423)
FEATURES             Location/Qualifiers
     source          1..7503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        1391..1409
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1391..1408
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      4146..4155
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4152..4154,4155..4157,4158..4871)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4152..4154,4155..4157,4158..4868)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4197..4218)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4458..4477)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4805..4826
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4994..5115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5031..5050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5085..5104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5122..5577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5259..5280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5471..5490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5604..5708
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5710..6067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5710..5901
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5733..5753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5918..6053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6102..6896
                     /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             6102..6896
                     /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(6156..6175)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6766..6785
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7084..7103)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7128..7175
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga
     1321 gctcaagctt ggtaccgagc tcggatctgc cttacaatta atacataacc ttttgtatca
     1381 tacacatacg atttaggtga cactatagaa ttcgggccgg cctaacccgg ccctgcccga
     1441 ccgcacccga gctcagtgtt tgctcggcgt ctgccgggtc cgccatggga gcccggtgcc
     1501 gcagcttttc cgcgctcctg ctcctgctgc aggtctcctc atggctttgc caggagctgg
     1561 agcctgagtc ctgcagtccc ggcttcagtt ccgaggtcta caccttcccg gtgccggaga
     1621 ggcacctgga gagaggccat gtcctgggca gagtgagatt tgaaggatgc accggccggc
     1681 caaggacagc cttcttttcg gaagactccc gattcaaagt ggcgacagac ggcaccatca
     1741 cagtgaagcg gcatctaaag ctccacaagc tggagaccag tttcctcgtc cgcgcccggg
     1801 actccagtca tagggagctg tctaccaaag tgacgctgaa gtccatgggg caccaccatc
     1861 accggcacca ccaccgcgac cctgcctctg aatccaaccc agagctgctc atgtttccca
     1921 gcgtgtaccc aggtctcaga agacagaaac gagactgggt catccctccc atcagctgcc
     1981 ccgaaaatga aaagggtgaa ttcccaaaga acctggttca gatcaaatcc aacagggaca
     2041 aagaaacaaa ggttttctac agcatcaccg gccaaggagc tgacaaaccc cccgttggcg
     2101 ttttcatcat tgagagggag acaggctggc tgaaagtgac acagcctctg gatagagaag
     2161 ccattgccaa gtacatcctc tattctcatg ccgtgtcatc aaatggggaa gcggtggagg
     2221 atcccatgga gatagtgatc acagtgacag atcagaatga caacaggcca gagtttaccc
     2281 aggaggtgtt tgagggatcc gttgcagaag gcgctgttcc aggaacctcc gtgatgaagg
     2341 tctcagccac cgatgcagac gatgacgtca acacctacaa cgctgccatc gcctacacca
     2401 tcgtcagcca ggatcctgag ctgcctcaca aaaacatgtt cactgtcaat agggacaccg
     2461 gggtcatcag tgtgctcacc tctgggctgg accgagagag ttaccctaca tacactctgg
     2521 tggttcaggc tgctgacctt caaggcgaag gcttgagcac aacagccaag gctgtgatca
     2581 ctgtcaagga tattaatgac aacgctcctg tcttcaaccc gagcacgtat cagggtcaag
     2641 tgcctgagaa tgaggtcaat gcccggatcg ccacactcaa agtgaccgat gatgatgccc
     2701 ccaacactcc ggcgtggaaa gctgtgtaca ccgtagtcaa cgatcctgac cagcagttcg
     2761 ttgtcgtcac agaccccacg accaatgatg gcattttgaa aacagccaag ggcttggatt
     2821 ttgaggccaa gcagcaatac atccttcatg tgagagtgga gaacgaggaa ccctttgagg
     2881 ggtctcttgt cccttccaca gccactgtca ctgtggacgt ggtagacgtg aatgaagccc
     2941 ccatctttat gcctgcggag aggagagtcg aagtgcccga agactttggt gtgggtcagg
     3001 aaatcacatc ttataccgct cgagagccgg acacgttcat ggatcagaag atcacgtatc
     3061 ggatttggag ggacactgcc aactggctgg agattaaccc agagactggt gccattttca
     3121 cgcgcgctga gatggacaga gaagacgctg agcatgtgaa gaacagcaca tatgtagctc
     3181 tcatcatcgc cacagatgat ggttcaccca ttgccactgg cacgggcact cttctcctgg
     3241 tcctgttaga cgtcaatgac aacgctccca tcccagaacc tcgaaacatg cagttctgcc
     3301 agaggaaccc acagcctcat atcatcacca tcttggatcc agaccttccc cccaacacgt
     3361 ccccctttac tgctgagcta acccatgggg ccagcgtcaa ctggaccatt gagtataatg
     3421 acgcagctca agaatctctc attttgcaac caagaaagga cttagagatt ggcgaataca
     3481 aaatccatct caagctcgcg gataaccaga acaaagacca ggtgaccacg ttggacgtcc
     3541 atgtgtgtga ctgtgaaggg acggtcaaca actgcatgaa ggcgggaatc gtggcagcag
     3601 gattgcaagt tcctgccatc ctcggaatcc ttggagggat cctcgccctg ctgattctga
     3661 tcctgctgct cctactgttt ctacggagga gaacggtggt caaagagccc ctgctgccac
     3721 cagatgatga tacccgggac aatgtgtatt actatgatga agaaggaggt ggagaagaag
     3781 accaggactt tgatttgagc cagctgcaca ggggcctgga tgcccgaccg gaagtgactc
     3841 gaaatgatgt ggctcccacc ctcatgagcg tgccccagta tcgtccccgt cctgccaatc
     3901 ctgatgaaat tggaaacttc atcgatgaaa acctgaaggc agccgacagc gaccccacgg
     3961 caccccctta cgactctctg ttggtgttcg attacgaggg cagtggttct gaagccgcta
     4021 gcctgagctc actgaactcc tctgagtcgg atcaggacca ggactacgat tatctgaacg
     4081 agtggggcaa ccgattcaag aagctggcgg acatgtacgg cggtggcgag gacgacaaac
     4141 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     4201 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     4261 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     4321 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     4381 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     4441 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     4501 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     4561 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     4621 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     4681 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     4741 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     4801 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     4861 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     4921 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4981 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5041 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5101 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     5161 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     5221 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     5281 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     5341 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     5401 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     5461 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     5521 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     5581 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5641 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5701 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     5761 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5821 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5881 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5941 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6001 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6061 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     6121 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     6181 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     6241 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     6301 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     6361 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     6421 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     6481 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     6541 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     6601 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     6661 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6721 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6781 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6841 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6901 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6961 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     7021 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     7081 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     7141 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     7201 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     7261 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     7321 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     7381 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7441 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7501 ttc
//
