LOCUS       sequence-351550-        7429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express alpha-catenin with M319G/R326E  mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7429)
  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 7429)
  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 M319G/R326E
FEATURES             Location/Qualifiers
     source          1..7429
                     /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    4217..4338
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4254..4273)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4308..4327
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4345..4800)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4482..4503)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4694..4713
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4827..4931
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4933..5290
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4933..5124
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4956..4976)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5141..5276
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5325..6119
                     /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             5325..6119
                     /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(5379..5398)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5989..6008
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6307..6326)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6351..6398
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6727..7315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7216..7235
                     /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 tgctgtccat gcaggcaaca taaacttcaa gtgggatcct
     1381 aaaagtctag agatcaggac tctggcagtt gagagactgt tggagcctct tgttacacag
     1441 gttacaaccc ttgtaaacac caatagtaaa gggccctcta ataagaagag aggtcgttct
     1501 aagaaggccc atgttttggc tgcatctgtt gaacaagcaa ctgagaattt cttggagaag
     1561 ggggataaaa ttgcgaagga gagccagttt ctcaaggagg agcttgtggc tgctgtagaa
     1621 gatgttcgaa aacaaggtga tttgatgaag gctgctgcag gagagttcgc agatgatccc
     1681 tgctcttctg tgaagcgagg caacatggtt cgggcagctc gagctttgct ctctgctgtt
     1741 acccggttgc tgattttggc tgacatggca gatgtctaca aattacttgt tcagctgaaa
     1801 gttgtggaag atggtatctt gaagttgagg aatgctggca atgaacaaga cttaggaatc
     1861 cagtataaag ccctaaaacc tgaagtggat aagctgaaca ttatggcagc caaaagacaa
     1921 caggaattga aagatgttgg ccatcgtgat cagatggctg cagctagagg aatcctgcag
     1981 aagaacgttc cgatcctcta tactgcatcc caggcatgcc tacagcaccc tgatgtcgca
     2041 gcctataagg ccaacaggga cctgatatac aagcagctgc agcaggcggt cacaggcatt
     2101 tccaatgcag cccaggccac tgcctcagac gatgcctcac agcaccaggg tggaggagga
     2161 ggagaactgg catatgcact caataacttt gacaaacaaa tcattgtgga ccccttgagc
     2221 ttcagcgagg agcgctttag gccttccctg gaggagcgtc tggaaagcat cattagtggg
     2281 gctgccttgg gcgccgactc gtcctgcacg gaggatgacc gtcgtgagcg aattgtggca
     2341 gagtgtaatg ctgtccgcca ggccctgcag gacctgcttt cggagtacat gggcaatgct
     2401 ggacgtaaag aaagaagtga tgcactcaat tctgcaatag ataaaatgac caagaagacc
     2461 agggacttgc gtagacagct ccgcaaagct gtcatggacc acgtttcaga ttctttcctg
     2521 gaaaccaatg ttccactttt ggtattgatt gaagctgcaa agaatggaaa tgagaaagaa
     2581 gttaaggagt atgcccaagt tttccgtgaa catgccaaca aattgattga ggttgccaac
     2641 ttggcctgtt ccatctcaaa taatgaagaa ggtgtaaagc ttgttcgaat gtctgcaagc
     2701 cagttagaag ccctctgtcc tcaggttatt aatgctgcac tggctttagc agcaaaacca
     2761 cagagtaaac tggcccaaga gaacatggat ctttttaaag aacaatggga aaaacaagtc
     2821 cgtgttctca cagatgctgt cgatgacatt acttccattg atgacttctt ggctgtctca
     2881 gagaatcaca ttttggaaga tgtgaacaaa tgtgtcattg ctctccaaga gaaggatgtg
     2941 gatggcctgg accgcacagc tggtgcaatt cgaggccggg cagcccgggt cattcacgta
     3001 gtcacctcag agatggacaa ctatgagcca ggagtctaca cagagaaggt tctggaagcc
     3061 actaagctgc tctccaacac agtcatgcca cgttttactg agcaagtaga agcagccgtg
     3121 gaagccctca gctcggaccc tgcccagccc atggatgaga atgagtttat cgatgcttcc
     3181 cgcctggtat atgatggcat ccgggacatc aggaaagcag tgctgatgat aaggacccct
     3241 gaggagttgg atgactctga ctttgagaca gaagattttg atgtcagaag caggacgagc
     3301 gtccagacag aagacgatca gctgatagct ggccagagtg cccgggcgat catggctcag
     3361 cttccccagg agcaaaaagc gaagattgcg gaacaggtgg ccagcttcca ggaagaaaag
     3421 agcaagctgg atgctgaagt gtccaaatgg gacgacagtg gcaatgacat cattgtgctg
     3481 gccaagcaga tgtgcatgat tatgatggag atgacagact ttacccgagg taaaggacca
     3541 ctcaaaaata catcggatgt catcagtgct gccaagaaaa ttgctgaggc aggatccagg
     3601 atggacaagc ttggccgcac cattgcagac cattgccccg actcggcttg caagcaggac
     3661 ctgctggcct acctgcaacg catcgccctc tactgccacc agctgaacat ctgcagcaag
     3721 gtcaaggccg aggtgcagaa tctcggcggg gagcttgttg tctctggggt ggacagcgcc
     3781 atgtccctga tccaggcagc caagaacttg atgaatgctg tggtgcagac agtgaaggca
     3841 tcctacgtcg cctctaccaa ataccaaaag tcacagggta tggcttccct caaccttcct
     3901 gctgtgtcat ggaagatgaa ggcaccagag aaaaagccat tggtgaagag agagaaacag
     3961 gatgagacac agaccaagat taaacgggca tctcagaaga agcacgtgaa cccggtgcag
     4021 gccctcagcg agttcaaagc tatggacagc atctaagaat tctgcagtcg acggtaccgc
     4081 gggcccggga tccaccggat ctagataact gatcataatc agccatacca catttgtaga
     4141 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4201 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4261 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4321 actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     4381 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4441 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4501 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4561 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4621 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4681 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4741 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4801 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4861 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4921 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     4981 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5041 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5101 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5161 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5221 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5281 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5341 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5401 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5461 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     5521 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5581 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5641 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5701 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5761 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5821 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     5881 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5941 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6001 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6061 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6121 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6181 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6241 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     6301 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     6361 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     6421 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     6481 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     6541 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     6601 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6661 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6721 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6781 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6841 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6901 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6961 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7021 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7081 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7141 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7201 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7261 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7321 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7381 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
