LOCUS       sequence-176133-        6242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasma membrane-targeted Venus (YFP)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6242)
  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 6242)
  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-Venus-CAAX
FEATURES             Location/Qualifiers
     source          1..6242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..18
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             22..54
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(58..66,67..81)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     regulatory      88..97
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(94..96,97..99,100..810)
                     /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(139..160)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(400..419)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     747..768
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(874..892)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(875..892)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(912..929)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    918..1142
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1188..1616
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1275..1294)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1485..1506
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1625..1645)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1630..1959
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1810..1945
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1872..1891
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2026..2820
                     /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(2080..2099)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2690..2709
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2994..3115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3031..3050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3085..3104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3164..3180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3164..3180)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3177..3199)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3188..3204
                     /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(3212..3218,3219..3236,3237..3242))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3257..3278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3395..3412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3566..4154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3646..3665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4325..5116,5117..5185))
                     /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     4948..4967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5186..5290)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5365..5384)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5556..5935
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5936..6139
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6090..6110
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6184..6203
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6184..6202
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      6222..6231
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 catcatcatc atcatcatgg tatggctagc atgactggtg gacagcaaat gggtcgggat
       61 ctgtacgacg atgacgataa ggatcccgcc accatggtga gcaagggcga ggagctgttc
      121 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      181 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
      241 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg
      301 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      361 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
      421 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
      481 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
      541 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
      601 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
      661 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc caagctgagc
      721 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
      781 atcactctcg gcatggacga gctgtacaag gaattcaaga agaagaagaa gagcaagacc
      841 aagtgcgtga tcatgtaatc tagagggccc tattctatag tgtcacctaa atgctagagc
      901 tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
      961 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1021 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1081 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1141 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1201 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1261 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1321 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1381 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1441 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1501 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1561 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     1621 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1681 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     1741 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     1801 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     1861 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     1921 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     1981 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2041 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2101 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2161 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2221 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2281 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2341 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2401 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2461 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     2521 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     2581 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     2641 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     2701 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     2761 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     2821 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     2881 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     2941 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3001 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3061 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3121 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3181 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3241 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3301 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3361 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3421 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3481 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3541 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3601 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3661 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3721 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3781 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3841 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3901 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3961 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4021 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4081 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4141 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4201 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4261 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4321 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4381 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4441 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4501 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4561 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4621 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4681 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4741 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4801 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4861 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4921 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4981 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5041 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5101 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5161 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5221 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5281 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     5341 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     5401 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     5461 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     5521 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     5581 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     5641 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     5701 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     5761 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     5821 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     5881 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     5941 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     6001 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     6061 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     6121 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
     6181 aattaatacg actcactata gggagaccca agcttgcggc cgccaccatg gtgcggggtt
     6241 ct
//
