LOCUS       sequence-118095-        7979 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a fusion of mVenus with MKL2 (MRTF-B) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7979)
  AUTHORS   van Unen J, Reinhard NR, Yin T, Wu YI, Postma M, Gadella TW, 
            Goedhart J
  TITLE     Plasma membrane restricted RhoGEF activity is sufficient for 
            RhoA-mediated actin polymerization.
  JOURNAL   Sci Rep. 2015 Oct 5;5:14693. doi: 10.1038/srep14693.
  PUBMED    26435194
REFERENCE   2  (bases 1 to 7979)
  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: "Sci Rep."; 
            date: "2015-10-5"; volume: "5:14693. doi"; pages: " 
            10.1038/srep14693"
COMMENT     Sequence Label: pmVenus-C2-MmMKL2
FEATURES             Location/Qualifiers
     source          1..7979
                     /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             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4591..4643
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    4767..4888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4804..4823)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4858..4877
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4895..5350)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5032..5053)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5244..5263
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5377..5481
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5483..5840
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5483..5674
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5506..5526)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5691..5826
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5875..6669
                     /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             5875..6669
                     /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(5929..5948)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6539..6558
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6857..6876)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6901..6948
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7277..7865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7766..7785
                     /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 gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag ctaccagtcc aagctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggccggac tcagatccag atgatcgata gctccaagaa gcagccacaa
     1381 ggcttcccag aaattttaac tgctgaggat tttgaacctt tcaaagaaaa ggaatgcctt
     1441 gagggaagca accagaaaag tctcaaggaa gtgctccagc tgaggctgca acaaaggagg
     1501 acaagagaac aactagtgga ccagggcatc atgccacctt tgaagagtcc agcagcattc
     1561 catgagcaga taaaaagcct ggagcgagcc agaactgaga attttctgaa gcacaagatc
     1621 cggagccgcc cagaccgctc tgagcttgtc aggatgcata ttttagaaga aacgtttgca
     1681 gagccatctc ttcaggccac acagatgaag ctgaagagag ctcggctggc agatgacctg
     1741 aacgagaaga ttgctcaaag gcctggtccc atggagctgg tggagaagaa catccttcct
     1801 gtggattcga gtgtcaagga agcgattata ggtgtggtga aggaggacta tcctcacact
     1861 catggcgagt tctcatttga tgaagatagc agtgatgctt tgtctccaga ccagccagcg
     1921 agccaggagt ctcagggctc agctgcatcc cccagtgagc caaaagtcag tgcctcgcca
     1981 cctcctgtga ctgcaagcac tccagcccag tttacctcag tgtccccagc agttcctgaa
     2041 ttcttgaaaa cccctctaac tgcagaccag cctcccacaa ggtctacagc tcccgtgctc
     2101 cctacaaaca ctgtgtcctc agcgaagtct ggcccaatgc tggtgaagca aagccatccc
     2161 aagaatccaa acgacaaaca ccgtagcaaa aaatgcaaag atccgaaacc gcgagtgaag
     2221 aagcttaagt accaccagta cattccaccc aaccagaagg gggagaagag cgagccacag
     2281 atggactcca actatgcccg cctgctccag cagcagcagc tcttcctaca gctgcagatc
     2341 ctgagccagc agcagcagca acagcagcag cagcactaca actaccagac catcctacct
     2401 gcacccatca aaactgacaa gaacagcagc agcggcagta acagcggcag tagcagcagc
     2461 atgccggcga ggaggccagg accgctgccc tccagcttgg atgacttaaa ggtgtcagag
     2521 ctgaagacag aactcaaact gaggggtctg ccagtgtcag gaactaaacc agacctcatt
     2581 gaacgcctga aaccctacca ggaagtgacc agtagtaacc ttgccactgg cagtattgtg
     2641 gcagtgtcat cagcaaccat tgtcaccagt aacccagagg tcacagtggc actgccagtc
     2701 acaacactgc ataatgctgt aacaagctct gtgtccactt tcaaggcaga cttggcactt
     2761 cctgcaacca gcagtgtgcc ccatgttgaa aatgcccact ctcctctgcc catctcccca
     2821 tcaccctctg agcagtccag tctcagtacc gatgacacaa acatgacaga caccttcaca
     2881 gagatcatga ctatgatgtc accttcccag ttgttgtgtt catctcccct gagggtggtg
     2941 agccatgatg acagcctgag cccttccagt agcaccctgt ctactctaga actggatgct
     3001 gctgagaagg accgcaaact tcaggagaag gagaaacaga ttgaagaact gaagaggaaa
     3061 ctggaacaag aacagaagct tgtggaggtc ctcaagatgc agcttgaggt ggaaaaacgt
     3121 gggcagcagc gaccaccaga ccctcagccc agtgatcccc cacatccctt taatacatca
     3181 gatcctaaac atggcagtgt tgggtcctcc atcaaggatg aggcctcact ccctgactgc
     3241 tccagccccc agcagcccat cacagtgcct ggccactctg taggccagcc catctccacg
     3301 ggtagccaga cccttgttgc caaaaagact gtggtcgtca agcaagaggt ccccatggcc
     3361 caagcagagc agcagaacgt tgtctcgcag ttctacctta gttcccaggg acagccacct
     3421 gcccttgttg ctcagcctca ggccttactg accacacaga caactcagct gctgctgcca
     3481 gtgtccatcc agggctcaaa tgttacttca gtgcaactcc cagtaggcag ccttcaactc
     3541 cagaccccag cacaaggaag agtccaggct cagcctcatg tagctgctgc cacacaggtt
     3601 ccagctgcag ccctgccctc agccttgacc tcagcactgc ctcagaagca ggaagccttc
     3661 ccacagcacg tgttgggcca gcctcagcca gtcagaaagg tcttcacaaa ctcggcacca
     3721 aacacagttc ttcagtatca gaggcagcct ggcccaacga accagcagcc ctttgtcagt
     3781 aagacctcta accctgctct tcaatccaga actgccccac ttgcacccct gcaaaatgga
     3841 cccagcctag ccagcaagcc tagctcacca cccccacccc agcagtttgt tgttcagcac
     3901 tctttgtttg caaccccaat cacgaagaca aaagatccac ctcgatatga ggaagccatc
     3961 aagcaggcac gtagcacaca gccagccctt ccagaggttt ccagtgtgca cagtcagcag
     4021 atggacgacc tcttcgacat cctcataaag agtggagaga tttccttccc aataaaagaa
     4081 gagccttctc caatttccaa gatgaaacca gtgacagcca gcatcaccac aatgccagtc
     4141 aataccgtgg tctctcgacc accgccccag gtccaaatag caccacctgt atccttagaa
     4201 cctgtgaaca gtttatctgc cagcttagag aaccagctag aagccttctt ggatggcact
     4261 ttgccctcgg ccactgacac tggcccactg cagaacagca gtgaagatag agagtctttc
     4321 tctctgatcg aggatctgca gaacgacttg ctgagtcatt ccagcatgct gtaccagtct
     4381 cactctccca tggagacctc tgaggcccag ttggtgtcag ggaccccctg tctatctctt
     4441 gacctgtcag actccaacct ggacaacatg gagtggctag acattaccat gcccaccaca
     4501 tcctccgggc tcactccact gagtaccact gcgccaagca tgttctctgc tgattttctg
     4561 gacccacagg acctgccgct gccatgggac ctcgagctca agcttcgaat tctgcagtcg
     4621 acggtaccgc gggcccggga tccaccggat ctagataact gatcataatc agccatacca
     4681 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4741 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4801 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4861 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     4921 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4981 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     5041 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     5101 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     5161 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     5221 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5281 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5341 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5401 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5461 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     5521 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5581 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5641 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5701 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5761 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5821 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5881 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5941 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     6001 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     6061 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     6121 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     6181 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6241 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6301 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6361 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     6421 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6481 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6541 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6601 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6661 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6721 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6781 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6841 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6901 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6961 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     7021 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     7081 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     7141 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7201 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7261 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7321 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7381 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     7441 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7501 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7561 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7621 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7681 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7741 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7801 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7861 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7921 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
