LOCUS       sequence-351548-        7420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express alpha-catenin without a vinculin binding site (VBS).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7420)
  AUTHORS   Bejar-Padilla V, Cabe JI, Lopez S, Narayanan V, Mezher M, 
            Maruthamuthu V, Conway DE
  TITLE     alpha-catenin Dependent Vinculin Recruitment to Adherens Junctions 
            is Antagonistic to Focal Adhesions.
  JOURNAL   Mol Biol Cell. 2022 Aug 3:mbcE22020071. doi: 
            10.1091/mbc.E22-02-0071.
  PUBMED    35921161
REFERENCE   2  (bases 1 to 7420)
  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: "Mol Biol 
            Cell. 2022 Aug 3:mbcE22020071. doi: 10.1091/mbc.E22-02-0071."
COMMENT     Sequence Label: pmcherry alpha-catenin Delta-VBS
FEATURES             Location/Qualifiers
     source          1..7420
                     /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             613..1320
                     /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(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4208..4329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4245..4264)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4299..4318
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4336..4791)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4473..4494)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4685..4704
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4818..4922
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4924..5281
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4924..5115
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4947..4967)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5132..5267
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5316..6110
                     /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             5316..6110
                     /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(5370..5389)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5980..5999
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6298..6317)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6342..6389
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6718..7306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7207..7226
                     /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 gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctatgac agccgtgcac gccggcaaca tcaacttcaa gtgggacccc
     1381 aagagcctgg aaatcagaac cctggccgtg gaaagactgc tggaacccct ggtcacccaa
     1441 gtgaccacac tggtcaacac caacagcaag ggccccagca acaagaagcg gggcagatcc
     1501 aaaaaggccc atgtgctggc tgccagcgtg gaacaggcca ccgagaactt tctggaaaag
     1561 ggcgacaaga tcgccaaaga gagccagttc ctgaaagagg aactggtcgc cgccgtggaa
     1621 gatgtgcgaa aacagggcga tctgatgaag gccgctgccg gcgaatttgc cgatgatcct
     1681 tgtagcagcg tgaagagggg caacatggtt cgagctgcta gagccctgct gagcgccgtg
     1741 actagactgc tgattctggc cgacatggcc gacgtgtaca agctgctggt gcagctgaag
     1801 gtggtcgagg acggaatcct gaagctgcgg aatgccggca acgagcagga tctgggcatc
     1861 cagtacaagg ccctgaagcc tgaggtggac aagctgaaca tcatggccgc caagcggcag
     1921 caagagctga aggatgtggg ccacagagat cagatggctg ccgccagagg catcctgcag
     1981 aaaaacgtgc ccatcctgta caccgccagc caggcttgtc tgcagcatcc tgatgtggcc
     2041 gcctacaagg ccaaccggga cctgatctat aagcagctgc agcaggccgt gaccggcatc
     2101 tctaatgctg ctcaggccac agcctccgac gatgcctctc aacatcaagg tggcggaggc
     2161 ggcgaactgg cctacgctct gaacaacttc gacaagcaga tcatcgtgga ccctctgagc
     2221 ttcagcgagg aacggttcag acccagcctg gaagagagac tggaatccat catctctggc
     2281 gccgagggac acagactggc caatgtgatg atgggcccct acagacagga cctgctggcc
     2341 aagtgcgaca gagtggatca gctgacagcc cagctggctg accttgctgc aagaggcgaa
     2401 ggcgaaagcc ctcaggctag agcacttgct agccagctgc aggacagcct gaaggacctg
     2461 aaggcccgga tgcaagaagc catgacacaa gaggtgtccg acgtgttcag cgacaccacc
     2521 acacctatca aactgctggc cgaggccgcc aaaaacggca atgagaaaga agtgaaagaa
     2581 tacgcccagg tgttccgcga gcacgccaac aagctgatcg aggtggccaa tctggcctgc
     2641 tccatcagca acaacgaaga gggcgtgaag ctcgtgcgga tgtctgcctc tcagctggaa
     2701 gccctgtgtc ctcaagtcat caatgccgct ctggccctgg ccgctaagcc tcagtctaaa
     2761 ctggcccaag agaacatgga cctgttcaaa gagcagtggg agaagcaagt gcgggtgctg
     2821 acagacgccg tggacgacat taccagcatc gacgactttc tggccgtgtc cgagaaccac
     2881 attctcgagg acgtgaacaa atgcgtgatc gccctccaag agaaggacgt ggacggcctg
     2941 gatagaacag ccggcgctat tagaggcaga gccgctagag tgatccacgt cgtgaccagc
     3001 gagatggaca actacgagcc tggcgtgtac accgagaagg tgctggaagc tacaaagctg
     3061 ctgagcaaca ccgtgatgcc ccggtttacc gaacaggtgg aagctgctgt ggaagctctg
     3121 agcagcgatc ccgctcagcc catggacgag aacgagttca tcgacgccag cagactggtg
     3181 tacgacggca tccgggatat cagaaaggcc gtgctgatga tcaggacccc tgaagaactg
     3241 gacgacagcg acttcgagac agaggacttc gacgtgcgga gcagaaccag cgtgcagacc
     3301 gaagatgacc agctgatcgc cggacagtct gccagagcta tcatggccca gctgcctcaa
     3361 gagcagaagg ccaagatcgc tgagcaggtc gccagcttcc aagaggaaaa gagcaagctg
     3421 gacgccgagg tgtccaagtg ggatgacagc ggcaacgaca tcatcgtgct ggctaagcag
     3481 atgtgtatga tcatgatgga aatgaccgac ttcaccagag gcaaggggcc tctgaagaac
     3541 accagcgacg tgatctctgc cgccaagaag attgccgaag ccggcagcag aatggataag
     3601 ctgggcagaa caatcgccga tcactgccct gatagcgcct gcaaacagga tctgctcgcc
     3661 tacctgcagc ggattgccct gtattgccac cagctgaata tctgcagcaa agtgaaggcc
     3721 gaggtgcaga atctcggcgg cgagcttgtt gtcagcggag tggattctgc catgagcctg
     3781 atccaggctg ccaagaacct gatgaacgct gtggtgcaga ctgtgaaggc cagctacgtg
     3841 gccagcacca agtaccagaa aagccaaggc atggccagcc tgaacctgcc tgccgtgtct
     3901 tggaaaatga aggcccctga gaagaagcct ctggtcaaga gagagaagca ggacgagaca
     3961 cagaccaaga tcaagcgggc cagccagaaa aagcacgtga accctgtgca ggccctgagc
     4021 gagttcaagg ccatggacag catctgagaa ttctgcagtc gacggtaccg cgggcccggg
     4081 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     4141 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4201 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4261 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4321 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4381 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4441 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4501 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4561 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4621 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4681 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4741 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     4801 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4861 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4921 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4981 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5041 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5101 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5161 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5221 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5281 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5341 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5401 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5461 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     5521 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5581 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5641 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5701 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5761 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     5821 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     5881 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     5941 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6001 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6061 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6121 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     6181 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6241 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     6301 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     6361 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     6421 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     6481 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     6541 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     6601 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6661 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6721 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6781 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6841 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     6901 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6961 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7021 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7081 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     7141 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7201 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7261 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7321 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7381 atcccctgat tctgtggata accgtattac cgccatgcat
//
