LOCUS       sequence-176218-        6197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Actin-targeted Venus (YFP)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6197)
  AUTHORS   Hertel F, Mo GC, Duwe S, Dedecker P, Zhang J
  TITLE     RefSOFI for Mapping Nanoscale Organization of Protein-Protein 
            Interactions in Living Cells.
  JOURNAL   Cell Rep. 2016 Jan 12;14(2):390-400. doi: 
            10.1016/j.celrep.2015.12.036. Epub 2015 Dec 31.
  PUBMED    26748717
REFERENCE   2  (bases 1 to 6197)
  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.celrep.2015.12.036"; journalName: "Cell Rep"; date: 
            "2016-01-12- 12"; volume: "14"; issue: "2"; pages: "390-400"
COMMENT     Sequence Label: pcDNA3-LifeAct-Venus
FEATURES             Location/Qualifiers
     source          1..6197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..51
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     regulatory      61..70
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(67..69,70..72,73..786)
                     /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(112..133)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(373..392)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     720..741
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(847..865)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(848..865)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(885..902)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    891..1115
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1161..1589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1248..1267)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1458..1479
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1598..1618)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1603..1932
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1783..1918
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1845..1864
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1999..2793
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2053..2072)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2663..2682
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2967..3088
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3004..3023)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3058..3077
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3137..3153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3137..3153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3150..3172)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3161..3177
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3185..3191,3192..3209,3210..3215))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3230..3251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3368..3385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3539..4127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3619..3638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4298..5089,5090..5158))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4921..4940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5159..5263)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5338..5357)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5529..5908
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5909..6112
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6063..6083
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6157..6176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6157..6175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      join(6192..6197,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atgggcgtgg ccgacctgat caagaagttc gagagcatca gcaaggagga gggggatccc
       61 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      121 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      181 acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg
      241 cccaccctcg tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac
      301 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      361 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      421 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
      481 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag
      541 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
      601 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      661 aaccactacc tgagctacca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
      721 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
      781 aagtaagaat tctgcagata tccatcacac tggcggccgc tcgagcatgc atctagaggg
      841 ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga ctgtgccttc
      901 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
      961 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1021 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     1081 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     1141 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1201 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1261 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1321 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1381 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1441 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1501 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1561 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     1621 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1681 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1741 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     1801 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     1861 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     1921 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     1981 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2041 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2101 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2161 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     2221 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2281 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2341 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     2401 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     2461 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     2521 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     2581 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     2641 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     2701 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     2761 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     2821 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     2881 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     2941 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     3001 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3061 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     3121 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3181 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3241 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3301 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3361 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3421 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3481 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3541 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3601 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3661 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3721 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3781 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3841 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3901 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3961 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4021 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4081 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4141 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4201 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4261 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4321 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4381 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4441 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4501 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4561 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4621 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4681 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4741 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4801 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4861 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4921 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4981 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5041 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5101 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5161 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5221 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5281 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     5341 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     5401 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     5461 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
     5521 tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     5581 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     5641 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     5701 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     5761 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     5821 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     5881 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     5941 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     6001 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     6061 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct
     6121 aactagagaa cccactgctt actggcttat cgaaattaat acgactcact atagggagac
     6181 ccaagcttgc cgccacc
//
