LOCUS       sequence-440941-        7943 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mouse vinculin with GFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7943)
  AUTHORS   Leerberg JM, Gomez GA, Verma S, Moussa EJ, Wu SK, Priya R, Hoffman 
            BD, Grashoff C, Schwartz MA, Yap AS
  TITLE     Tension-sensitive actin assembly supports contractility at the 
            epithelial zonula adherens.
  JOURNAL   Curr Biol. 2014 Aug 4;24(15):1689-99. doi: 
            10.1016/j.cub.2014.06.028. Epub 2014 Jul 24.
  PUBMED    25065757
REFERENCE   2  (bases 1 to 7943)
  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: 
            "10.1016/j.cub.2014.06.028"; journalName: "Curr Biol"; date: 
            "2014-08-4- 4"; volume: "24"; issue: "15"; pages: "1689-99"
COMMENT     Sequence Label: GFP-mouse vinculin full length (889)
FEATURES             Location/Qualifiers
     source          1..7943
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4671..4792
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4708..4727)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4762..4781
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4799..5254)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4936..4957)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5148..5167
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5281..5385
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5387..5744
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5387..5578
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5410..5430)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5595..5730
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5779..6573
                     /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             5779..6573
                     /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(5833..5852)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6443..6462
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6761..6780)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6805..6852
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7181..7769
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7670..7689
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attccgccac catgccggtg
     1321 tttcatacgc gtacgatcga gagcatcctg gagccggtgg cgcagcagat ctcgcacctg
     1381 gtgattatgc acgaggaggg cgaggtggac ggcaaagcca ttcctgacct caccgcgccc
     1441 gtagccgccg tgcaggcggc cgtcagcaac ctcgtccggg ttggaaaaga gactgttcag
     1501 accactgagg atcagattct gaagagagat atgccaccag cctttattaa ggttgaaaat
     1561 gcttgcacca agcttgttca ggcagcccag atgcttcagt cagacccata ctcggttcct
     1621 gcgcgggatt acctcattga cggctctagg ggaatccttt ctggcacatc tgacctactg
     1681 cttaccttcg atgaggctga ggttcgtaaa attattaggg tttgcaaagg aattttggaa
     1741 tatcttacag tggcagaggt agtggaaact atggaagact tggtcactta cacaaagaat
     1801 cttgggccag gaatgactaa gatggccaaa atgattgatg agagacagca ggagttgact
     1861 caccaggaac accgtgtgat gttggtgaac tcaatgaaca ctgtcaaaga gctgcttcct
     1921 gttctcattt cagctatgaa gatttttgtt acaaccaaaa actcaaaaaa ccaaggaata
     1981 gaagaagctt tgaaaaatcg aaattttact gtagaaaaga tgagtgctga aattaacgag
     2041 atcattcgtg tgttacaact cacttcctgg gatgaagatg cctgggccag caaggacact
     2101 gaagccatga agagagcgct ggcgtccata gactccaaat tgaaccaggc caaaggttgg
     2161 ctccgtgacc ccaatgcctc cccaggggat gctggagagc aggccatcag gcagatctta
     2221 gatgaagctg gaaaagttgg tgaactttgt gcaggcaagg aacgcaggga gatcctagga
     2281 acctgcaaaa tgctagggca gatgactgac caagtggctg acctccgagc cagaggacaa
     2341 ggagcttccc cagtggccat gcagaaggcc cagcaagtgt ctcaggggct cgacgtgctt
     2401 accgccaaag tggagaatgc agctcggaag ctggaagcca tgacgaactc aaagcagagc
     2461 attgcaaaga agattgatgc tgcccagaat tggctggcgg atccaaatgg tggacctgag
     2521 ggagaagaac agattcgagg ggctttggct gaagctcgga agattgcaga attatgtgat
     2581 gatcctaagg agagagatga catcctccgc tcccttggag agatagctgc tctgacctct
     2641 aaactaggag acttgcgaag acaggggaaa ggagactcgc cagaggctcg agccttggct
     2701 aaacaagtgg cgacggcact acagaacctg cagaccaaaa ccaacagggc cgtggccaac
     2761 agcagacctg ccaaagcagc tgtccacctc gagggcaaga ttgaacaggc gcagcggtgg
     2821 attgataacc ccacagtaga tgaccgtgga gtcggtcagg ctgccatccg tggacttgtg
     2881 gctgaggggc atcggctggc caatgtcatg atgggacctt atcgccaaga tcttcttgcc
     2941 aaatgtgacc gtgtagacca gctaacagct cagctggctg acctggctgc ccgaggggag
     3001 ggggagagtc ctcaggcgag agcacttgca tcccagcttc aggactcctt aaaggatctt
     3061 aaagcccaga tgcaggaagc tatgactcaa gaggtatccg atgttttcag cgatactaca
     3121 actcctatca agctgttggc agtagccgcc actgctcctc ctgatgcacc caatagggaa
     3181 gaggtatttg atgaaagggc agccaattct gaaaaccatt caggaaggct tggagccaca
     3241 gcagagaagg cggctgctgt tggtactgcc aataaatcga cagtggaagg cattcaggca
     3301 tctgtgaaga cagcccgaga actcactccc caggtcatct ccgctgctcg gatcttactg
     3361 aggaaccctg gtaaccaggc tgcttatgaa cattttgaga ccatgaagaa ccagtggatt
     3421 gataatgttg aaaaaatgac agggctggtg gacgaggcta ttgataccaa gtctctgttg
     3481 gatgcttctg aagaagcaat taaaaaagac ctggacaagt gtaaggtagc catggccaat
     3541 attcagcctc agatgctggt cgctggagca accagtattg ctcgtcgggc caaccggatt
     3601 ctgctggttg ctaagaggga ggtagagaac tctgaggacc cgaagttccg agaggctgtg
     3661 aaagctgcct ctgatgaact gagcaaaaca atctccccca tggtgatgga tgccaaggct
     3721 gtggctggaa acatctctga ccctggcctg caaaagagct tcctggactc aggatatcgg
     3781 atcctgggag ctgaggccaa ggtcagagaa gccttccaac ctcaggagcc tgacttcccg
     3841 cctcctccac cagaccttga acagctacga ctaactgatg agctggctcc tcctaagcca
     3901 cctctgcctg agggtgaagt ccctccaccc aggcccccac caccagaaga gaaggatgaa
     3961 gagttccctg agcagaaagc tggtgaggtg attaaccagc caatgatgat ggccgccagg
     4021 cagctccacg atgaagctcg gaaatggtct agcaagggca atgacatcat tgcagcagcc
     4081 aagcgcatgg ctctgctgat ggcagagatg tctcggccgg taagaggggg cagtggtacc
     4141 aagcgggcac ttattcagtg tgccaaggat atcgccaagg cctctgatga ggtgacgagg
     4201 ttggccaagg aggttgccaa gcagtgcaca gataagcgga ttagaaccaa tctcttacag
     4261 gtatgcgagc gaatcccaac tataagcacc cagctcaaaa tcctatccac agtgaaggcc
     4321 actatgctgg gccggaccaa catcagtgat gaggagtctg agcaggccac agagatgctg
     4381 gttcataatg cccagaacct catgcagtct gtgaaggaga ctgtgcgaga ggctgaagct
     4441 gcttcaatca aaatccgaac agatgctggc tttactctgc gctgggtcag aaagactccc
     4501 tggtaccagc gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat
     4561 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     4621 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     4681 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4741 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     4801 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     4861 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4921 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4981 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     5041 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     5101 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     5161 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     5221 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     5281 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5341 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     5401 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     5461 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     5521 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     5581 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     5641 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     5701 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     5761 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     5821 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     5881 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5941 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     6001 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     6061 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     6121 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     6181 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     6241 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     6301 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     6361 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     6421 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     6481 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     6541 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     6601 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     6661 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6721 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     6781 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     6841 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     6901 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     6961 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     7021 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     7081 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7141 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7201 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7261 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7321 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7381 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7441 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7501 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7561 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7621 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7681 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7741 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7801 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7861 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     7921 catagcccat atatggagtt ccg
//
