LOCUS       sequence-127487-        6384 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Entry clone containing three repeats of Venus. Can be fused with 
            linker to the C-terminal end of protein of interest. For use in 
            plants and compatible with the MultiSite Gateway system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6384)
  AUTHORS   Siligato R, Wang X, Yadav SR, Lehesranta S, Ma G, Ursache R, Sevilem
            I, Zhang J, Gorte M, Prasad K, Wrzaczek M, Heidstra R, Murphy A, 
            Scheres B, Mahonen AP
  TITLE     MultiSite Gateway compatible cell type-specific gene inducible 
            system for plants.
  JOURNAL   Plant Physiol. 2015 Dec 7. pii: pp.01246.2015.
  PUBMED    26644504
REFERENCE   2  (bases 1 to 6384)
  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: "Plant 
            Physiol. 2015 Dec 7. pii: pp.01246.2015."
COMMENT     Sequence Label: p2R3a-3xVenusYFP-OcsT
FEATURES             Location/Qualifiers
     source          1..6384
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(126..144)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(149..165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(246..264)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(282..298)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(295..317)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    306..322
                     /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(330..336,337..354,355..360))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    375..396
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(513..530)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(684..1272)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(764..783)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1443..2234,2235..2303))
                     /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     2066..2085
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2304..2408)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2486..2941)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2623..2644)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2835..2854
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3067..3089
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3082..3098
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3105..3123
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3154..3163
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3158..3167
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3183..3199
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3270..3393)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(3420..3422,3423..3425,3426..4136)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4143..4145,4146..4148,4149..4859)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4866..4868,4869..4871,4872..5582)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      5601..6307
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(6348..6366)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    complement(join(6377..6384,1..88))
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ttatacaaag ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
       61 actatcagtc aaaataaaat cattatttgg agctccatgg tagcgttaac gcggccgcga
      121 tatcccctat agtgagtcgt attacatggt catagctgtt tcctgtatca ctagtgaatt
      181 cgcggccgcc tgcaggtcga ccatatggga gagctcccaa cgcgttggat gcatagcttg
      241 agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg
      301 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      361 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      421 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      481 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1081 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1741 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2401 gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg
     2461 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg
     2521 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2581 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2641 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2701 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2761 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2821 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2881 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2941 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     3001 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     3061 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat
     3121 agggcgaatt gggcccgacg tcgcatgctc ccggccgcca tggcggccgc gggaattcga
     3181 ttgtaaaacg acggccagtc ttaagctcgg gccctgcagc tctagagctc gaattctaca
     3241 ggtcactaat accatctaag tagttgattc atagtgactg catatgttgt gttttacagt
     3301 attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt
     3361 acgtttctcg ttcaactttc ttgtacaaag tggggatcca agcttcgcag gctggatcca
     3421 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     3481 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     3541 gcaagctgac cctgaagctg atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3601 tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc
     3661 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3721 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3781 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3841 agctggagta caactacaac agccacaacg tctatatcac cgccgacaag cagaagaacg
     3901 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg
     3961 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     4021 acctgagcta ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     4081 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagagat
     4141 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     4201 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     4261 acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca
     4321 ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
     4381 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     4441 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     4501 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     4561 acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga
     4621 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     4681 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     4741 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     4801 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaaga
     4861 gatccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     4921 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     4981 cctacggcaa gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc
     5041 ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt cgcccgctac cccgaccaca
     5101 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     5161 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     5221 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     5281 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga
     5341 agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     5401 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     5461 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     5521 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     5581 agcatgatta actcgaggtc ctgctttaat gagatatgcg agacgcctat gatcgcatga
     5641 tatttgcttt caattctgtt gtgcacgttg taaaaaacct gagcatgtgt agctcagatc
     5701 cttaccgccg gtttcggttc attctaatga atatatcacc cgttactatc gtatttttat
     5761 gaataatatt ctccgttcaa tttactgatt gtaccctact acttatatgt acaatattaa
     5821 aatgaaaaca atatattgtg ctgaataggt ttatagcgac atctatgata gagcgccaca
     5881 ataacaaaca attgcgtttt attattacaa atccaatttt aaaaaaagcg gcagaaccgg
     5941 tcaaacctaa aagactgatt acataaatct tattcaaatt tcaaaaggcc ccaggggcta
     6001 gtatctacga cacaccgagc ggcgaactaa taacgttcac tgaagggaac tccggttccc
     6061 cgccggcgcg catgggtgag attccttgaa gttgagtatt ggccgtccgc tctaccgaaa
     6121 gttacgggca ccattcaacc cggtccagca cggcggccgg gtaaccgact tgctgccccg
     6181 agaattatgc agcatttttt tggtgtatgt gggccccaaa tgaagtgcag gtcaaacctt
     6241 gacagtgacg acaaatcgtt gggcgggtcc agggcgaatt ttgcgacaac atgtcgaggc
     6301 tcagcaggac ctgcaggcat gcaagctagc ttactagtga tgcatattct atagtgtcac
     6361 ctaaatctgc ggccgccaac ttta
//
