LOCUS       sequence-351549-        8198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express alpha-catenin with L344P mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8198)
  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 8198)
  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 L344P
FEATURES             Location/Qualifiers
     source          1..8198
                     /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    4986..5107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5023..5042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5077..5096
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5114..5569)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5251..5272)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5463..5482
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5596..5700
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5702..6059
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5702..5893
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5725..5745)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5910..6045
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6094..6888
                     /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             6094..6888
                     /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(6148..6167)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6758..6777
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7076..7095)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7120..7167
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7496..8084
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7985..8004
                     /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 ccggactcag atctcgagct caagcttcga attccggaaa tgactgctgt ccatgcaggc
     1381 aacataaact tcaagtggga tcctaaaagt ctagagatca ggactctggc agttgagaga
     1441 ctgttggagc ctcttgttac acaggttaca acccttgtaa acaccaatag taaagggccc
     1501 tctaataaga agagaggtcg ttctaagaag gcccatgttt tggctgcatc tgttgaacaa
     1561 gcaactgaga atttcttgga gaagggggat aaaattgcga aggagagcca gtttctcaag
     1621 gaggagcttg tggctgctgt agaagatgtt cgaaaacaag gtgatttgat gaaggctgct
     1681 gcaggagagt tcgcagatga tccctgctct tctgtgaagc gaggcaacat ggttcgggca
     1741 gctcgagctt tgctctctgc tgttacccgg ttgctgattt tggctgacat ggcagatgtc
     1801 tacaaattac ttgttcagct gaaagttgtg gaagatggta tcttgaagtt gaggaatgct
     1861 ggcaatgaac aagacttagg aatccagtat aaagccctaa aacctgaagt ggataagctg
     1921 aacattatgg cagccaaaag acaacaggaa ttgaaagatg ttggccatcg tgatcagatg
     1981 gctgcagcta gaggaatcct gcagaagaac gttccgatcc tctatactgc atcccaggca
     2041 tgcctacagc accctgatgt cgcagcctat aaggccaaca gggacctgat atacaagcag
     2101 ctgcagcagg cggtcacagg catttccaat gcagcccagg ccactgcctc agacgatgcc
     2161 tcacagcacc agggtggagg aggaggagaa ctggcatatg cactcaataa ctttgacaaa
     2221 caaatcattg tggacccctt gagcttcagc gaggagcgct ttaggccttc cctggaggag
     2281 cgtctggaaa gcatcattag tggggctgcc ttgatggccg actcgtcctg cacgcgtgat
     2341 gaccgtcgtg agcgaattgt ggcagagtgt aatgctgtcc gccaggcccc acaggacctg
     2401 ctttcggagt acatgggcaa tgctggacgt aaagaaagaa gtgatgcact caattctgca
     2461 atagataaaa tgaccaagaa gaccagggac ttgcgtagac agctccgcaa agctgtcatg
     2521 gaccacgttt cagattcttt cctggaaacc aatgttccac ttttggtatt gattgaagct
     2581 gcaaagaatg gaaatgagaa agaagttaag gagtatgccc aagttttccg tgaacatgcc
     2641 aacaaattga ttgaggttgc caacttggcc tgttccatct caaataatga agaaggtgta
     2701 aagcttgttc gaatgtctgc aagccagtta gaagccctct gtcctcaggt tattaatgct
     2761 gcactggctt tagcagcaaa accacagagt aaactggccc aagagaacat ggatcttttt
     2821 aaagaacaat gggaaaaaca agtccgtgtt ctcacagatg ctgtcgatga cattacttcc
     2881 attgatgact tcttggctgt ctcagagaat cacattttgg aagatgtgaa caaatgtgtc
     2941 attgctctcc aagagaagga tgtggatggc ctggaccgca cagctggtgc aattcgaggc
     3001 cgggcagccc gggtcattca cgtagtcacc tcagagatgg acaactatga gccaggagtc
     3061 tacacagaga aggttctgga agccactaag ctgctctcca acacagtcat gccacgtttt
     3121 actgagcaag tagaagcagc cgtggaagcc ctcagctcgg accctgccca gcccatggat
     3181 gagaatgagt ttatcgatgc ttcccgcctg gtatatgatg gcatccggga catcaggaaa
     3241 gcagtgctga tgataaggac ccctgaggag ttggatgact ctgactttga gacagaagat
     3301 tttgatgtca gaagcaggac gagcgtccag acagaagacg atcagctgat agctggccag
     3361 agtgcccggg cgatcatggc tcagcttccc caggagcaaa aagcgaagat tgcggaacag
     3421 gtggccagct tccaggaaga aaagagcaag ctggatgctg aagtgtccaa atgggacgac
     3481 agtggcaatg acatcattgt gctggccaag cagatgtgca tgattatgat ggagatgaca
     3541 gactttaccc gaggtaaagg accactcaaa aatacatcgg atgtcatcag tgctgccaag
     3601 aaaattgctg aggcaggatc caggatggac aagcttggcc gcaccattgc agaccattgc
     3661 cccgactcgg cttgcaagca ggacctgctg gcctacctgc aacgcatcgc cctctactgc
     3721 caccagctga acatctgcag caaggtcaag gccgaggtgc agaatctcgg cggggagctt
     3781 gttgtctctg gggtggacag cgccatgtcc ctgatccagg cagccaagaa cttgatgaat
     3841 gctgtggtgc agacagtgaa ggcatcctac gtcgcctcta ccaaatacca aaagtcacag
     3901 ggtatggctt ccctcaacct tcctgctgtg tcatggaaga tgaaggcacc agagaaaaag
     3961 ccattggtga agagagagaa acaggatgag acacagacca agattaaacg ggcatctcag
     4021 aagaagcacg tgaacccggt gcaggccctc agcgagttca aagctatgga cagcatctaa
     4081 gtctgcccag gccggccgcc cccacccctc tgggctcctg aatatcagtc actgttcgtc
     4141 actcaaatga atttgctaaa tacaacactg atactagatt ccacagggaa atgggcagac
     4201 tgaaccagtc caggtggtga attttccaag aacatagttt aagttgatta aaaatgcttt
     4261 tagaatgcag gagcctactt ctagctgtat tttttgtatg cttaaataaa aataaaaatt
     4321 cataaccaaa gagaatccca cattagcttg ttagtaatgc tctgaccaag ccgagatgcc
     4381 cattctctta gtgatggcgg cgttagggtt tgagagaagg gaatttggct caacttcagt
     4441 tgagagggtg cagtccagac agcttgactg cttttaaatg accaaagatg acctgtggta
     4501 agcaacctgg gcatcttagg aagcagtccc tggagaaggc atgttcccag aaaggtctct
     4561 ggagggacaa actcactcag taaaacataa tgtatcatga agaaaactga ttctctatga
     4621 catgaaatga aaattttaat gcattgttat aattactaat gtacgctgct gcaggacatt
     4681 aataaagttg cttttttagg ctacagtgtc tcgatgccat aatcagaaca cacttttttt
     4741 cctctttctc ccagcttcaa atgcaaattc atcattgggc tcacttctaa taactgcagt
     4801 gtttcccgcc ttgggcttgc agccggaatt ctgcagtcga cggtaccgcg ggcccgggat
     4861 ccaccggatc tagataactg atcataatca gccataccac atttgtagag gttttacttg
     4921 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     4981 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5041 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5101 tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5161 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5221 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5281 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5341 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5401 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5461 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5521 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     5581 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5641 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5701 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5761 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5821 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5881 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5941 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6001 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     6061 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     6121 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     6181 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     6241 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     6301 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6361 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6421 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6481 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6541 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6601 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     6661 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6721 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6781 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6841 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6901 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6961 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     7021 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     7081 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     7141 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     7201 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     7261 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     7321 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     7381 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7441 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7501 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7561 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7621 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7681 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7741 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7801 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7861 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7921 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7981 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8041 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8101 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     8161 cccctgattc tgtggataac cgtattaccg ccatgcat
//
