LOCUS       sequence-147999-        7494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of murine E-caderin with mCherry fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7494)
  AUTHORS   Han SP, Gambin Y, Gomez GA, Verma S, Giles N, Michael M, Wu SK, Guo 
            Z, Johnston W, Sierecki E, Parton RG, Alexandrov K, Yap AS
  TITLE     Cortactin scaffolds Arp2/3 and WAVE2 at the epithelial zonula 
            adherens.
  JOURNAL   J Biol Chem. 2014 Mar 14;289(11):7764-75. doi: 
            10.1074/jbc.M113.544478. Epub 2014 Jan 27.
  PUBMED    24469447
REFERENCE   2  (bases 1 to 7494)
  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: 
            "10.1074/jbc.M113.544478"; journalName: "J Biol Chem"; date: 
            "2014-03-14- 14"; volume: "289"; issue: "11"; pages: "7764-75"
COMMENT     Sequence Label: Murine E-cadherin mCherry
FEATURES             Location/Qualifiers
     source          1..7494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        695..713
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     695..712
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      3450..3459
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3456..4166
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3601..3619)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3869..3888
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4289..4410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4326..4345)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4380..4399
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4417..4872)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4554..4575)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4766..4785
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4899..5003
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5005..5362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5005..5196
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5028..5048)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5213..5348
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5397..6191
                     /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             5397..6191
                     /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(5451..5470)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6061..6080
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6379..6398)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6423..6470
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6799..7387
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7288..7307
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagctcaa gcttggtacc gagctcggat ctgccttaca
      661 attaatacat aaccttttgt atcatacaca tacgatttag gtgacactat agaattcggg
      721 ccggcctaac ccggccctgc ccgaccgcac ccgagctcag tgtttgctcg gcgtctgccg
      781 ggtccgccat gggagcccgg tgccgcagct tttccgcgct cctgctcctg ctgcaggtct
      841 cctcatggct ttgccaggag ctggagcctg agtcctgcag tcccggcttc agttccgagg
      901 tctacacctt cccggtgccg gagaggcacc tggagagagg ccatgtcctg ggcagagtga
      961 gatttgaagg atgcaccggc cggccaagga cagccttctt ttcggaagac tcccgattca
     1021 aagtggcgac agacggcacc atcacagtga agcggcatct aaagctccac aagctggaga
     1081 ccagtttcct cgtccgcgcc cgggactcca gtcataggga gctgtctacc aaagtgacgc
     1141 tgaagtccat ggggcaccac catcaccggc accaccaccg cgaccctgcc tctgaatcca
     1201 acccagagct gctcatgttt cccagcgtgt acccaggtct cagaagacag aaacgagact
     1261 gggtcatccc tcccatcagc tgccccgaaa atgaaaaggg tgaattccca aagaacctgg
     1321 ttcagatcaa atccaacagg gacaaagaaa caaaggtttt ctacagcatc accggccaag
     1381 gagctgacaa accccccgtt ggcgttttca tcattgagag ggagacaggc tggctgaaag
     1441 tgacacagcc tctggataga gaagccattg ccaagtacat cctctattct catgccgtgt
     1501 catcaaatgg ggaagcggtg gaggatccca tggagatagt gatcacagtg acagatcaga
     1561 atgacaacag gccagagttt acccaggagg tgtttgaggg atccgttgca gaaggcgctg
     1621 ttccaggaac ctccgtgatg aaggtctcag ccaccgatgc agacgatgac gtcaacacct
     1681 acaacgctgc catcgcctac accatcgtca gccaggatcc tgagctgcct cacaaaaaca
     1741 tgttcactgt caatagggac accggggtca tcagtgtgct cacctctggg ctggaccgag
     1801 agagttaccc tacatacact ctggtggttc aggctgctga ccttcaaggc gaaggcttga
     1861 gcacaacagc caaggctgtg atcactgtca aggatattaa tgacaacgct cctgtcttca
     1921 acccgagcac gtatcagggt caagtgcctg agaatgaggt caatgcccgg atcgccacac
     1981 tcaaagtgac cgatgatgat gcccccaaca ctccggcgtg gaaagctgtg tacaccgtag
     2041 tcaacgatcc tgaccagcag ttcgttgtcg tcacagaccc cacgaccaat gatggcattt
     2101 tgaaaacagc caagggcttg gattttgagg ccaagcagca atacatcctt catgtgagag
     2161 tggagaacga ggaacccttt gaggggtctc ttgtcccttc cacagccact gtcactgtgg
     2221 acgtggtaga cgtgaatgaa gcccccatct ttatgcctgc ggagaggaga gtcgaagtgc
     2281 ccgaagactt tggtgtgggt caggaaatca catcttatac cgctcgagag ccggacacgt
     2341 tcatggatca gaagatcacg tatcggattt ggagggacac tgccaactgg ctggagatta
     2401 acccagagac tggtgccatt ttcacgcgcg ctgagatgga cagagaagac gctgagcatg
     2461 tgaagaacag cacatatgta gctctcatca tcgccacaga tgatggttca cccattgcca
     2521 ctggcacggg cactcttctc ctggtcctgt tagacgtcaa tgacaacgct cccatcccag
     2581 aacctcgaaa catgcagttc tgccagagga acccacagcc tcatatcatc accatcttgg
     2641 atccagacct tccccccaac acgtccccct ttactgctga gctaacccat ggggccagcg
     2701 tcaactggac cattgagtat aatgacgcag ctcaagaatc tctcattttg caaccaagaa
     2761 aggacttaga gattggcgaa tacaaaatcc atctcaagct cgcggataac cagaacaaag
     2821 accaggtgac cacgttggac gtccatgtgt gtgactgtga agggacggtc aacaactgca
     2881 tgaaggcggg aatcgtggca gcaggattgc aagttcctgc catcctcgga atccttggag
     2941 ggatcctcgc cctgctgatt ctgatcctgc tgctcctact gtttctacgg aggagaacgg
     3001 tggtcaaaga gcccctgctg ccaccagatg atgatacccg ggacaatgtg tattactatg
     3061 atgaagaagg aggtggagaa gaagaccagg actttgattt gagccagctg cacaggggcc
     3121 tggatgcccg accggaagtg actcgaaatg atgtggctcc caccctcatg agcgtgcccc
     3181 agtatcgtcc ccgtcctgcc aatcctgatg aaattggaaa cttcatcgat gaaaacctga
     3241 aggcagccga cagcgacccc acggcacccc cttacgactc tctgttggtg ttcgattacg
     3301 agggcagtgg ttctgaagcc gctagcctga gctcactgaa ctcctctgag tcggatcagg
     3361 accaggacta cgattatctg aacgagtggg gcaaccgatt caagaagctg gcggacatgt
     3421 acggcggtgg cgaggacgac aaaccggtcg ccaccatggt gagcaagggc gaggaggata
     3481 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     3541 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     3601 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     3661 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     3721 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     3781 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     3841 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     3901 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     3961 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     4021 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     4081 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     4141 gcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac
     4201 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     4261 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     4321 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4381 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     4441 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4501 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4561 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4621 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4681 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4741 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4801 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4861 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4921 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4981 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     5041 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5101 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5161 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5221 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5281 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5341 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     5401 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5461 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5521 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     5581 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5641 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5701 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5761 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5821 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5881 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     5941 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6001 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6061 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6121 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6181 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6241 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6301 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6361 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     6421 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     6481 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     6541 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     6601 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     6661 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6721 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6781 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6841 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6901 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6961 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7021 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7081 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7141 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7201 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7261 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7321 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7381 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7441 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgc
//
