LOCUS       sequence-246311-        4474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for making C-terminal TurboID-mVenus fusion 
            with non-nuclear proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4474)
  AUTHORS   Mair A, Xu SL, Branon TC, Ting AY, Bergmann DC
  TITLE     Proximity labeling of protein complexes and cell type-specific 
            organellar proteomes in Arabidopsis enabled by TurboID.
  JOURNAL   Elife. 2019 Sep 19;8. pii: 47864. doi: 10.7554/eLife.47864.
  PUBMED    31535972
REFERENCE   2  (bases 1 to 4474)
  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: "Elife."; 
            date: "2019-09-19"; volume: "8. pii: 47864. doi"; pages: " 
            10.7554/eLife.47864"
COMMENT     Sequence Label: pDONR_P2R-P3_R2-Turbo-NES-mVenus-STOP-L3
FEATURES             Location/Qualifiers
     source          1..4474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             38..79
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             86..1042
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLG
                     ADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWI
                     TLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGDK
                     EIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             1049..1075
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             join(1115..1117,1118..1120,1121..1834)
                     /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"
     primer_bind     complement(1421..1440)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1768..1789
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1835..1930)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1967..1986)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1968..1986)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1991..2007)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1991..2007)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2109..2128)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2120..2929
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2187..2206)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3076..3664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3565..3584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3818..3835
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3994..4021)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4113..4199)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4248..4270
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4262..4279
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4263..4279
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4350..4473)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tgggcgcgcc ggtggaggcg gttcaggagg tggcatgggc aagcccatcc ccaaccccct
       61 gctgggcctg gacagcaccg ctagcaaaga caatactgtg cctctgaagc tgatcgctct
      121 cctggctaat ggcgagttcc atagtggcga acagctggga gaaaccctgg gcatgtccag
      181 ggccgctatc aacaagcaca ttcagactct gcgcgactgg ggcgtggacg tgttcaccgt
      241 gcccggaaag ggctactctc tgcccgagcc tatcccgctg ctgaacgcta aacagattct
      301 gggacagctg gacggcggga gcgtggcagt cctgcctgtg gtcgactcca ccaatcagta
      361 cctgctggat cgaatcggcg agctgaagag tggggatgct tgcattgcag aatatcagca
      421 ggcagggaga ggaagcagag ggaggaaatg gttctctcct tttggagcta acctgtacct
      481 gagtatgttt tggcgcctga agcggggacc agcagcaatc ggcctgggcc cggtcatcgg
      541 aattgtcatg gcagaagcgc tgcgaaagct gggagcagac aaggtgcgag tcaaatggcc
      601 caatgacctg tatctgcagg atagaaagct ggcaggcatc ctggtggagc tggccggaat
      661 aacaggcgat gctgcacaga tcgtcattgg cgccgggatt aacgtggcta tgaggcgcgt
      721 ggaggaaagc gtggtcaatc agggctggat cacactgcag gaagcaggga ttaacctgga
      781 caggaatact ctggccgcta cgctgatccg agagctgcgg gcagccctgg aactgttcga
      841 gcaggaaggc ctggctccat atctgccacg gtgggagaag ctggataact tcatcaatag
      901 acccgtgaag ctgatcattg gggacaaaga gattttcggg attagccggg ggattgataa
      961 acagggagcc ctgctgctgg aacaggacgg agttatcaaa ccctggatgg gcggagaaat
     1021 cagtctgcgg tctgccgaaa agctgcagct gcctcccctg gagcgcctga ccctggacgg
     1081 aggcggtgga agcggcggag gtgggcgcgc ctccatggtg agcaagggcg aggagctgtt
     1141 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1201 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg
     1261 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg gctacggcct
     1321 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1381 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1441 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1501 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1561 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcaagatccg
     1621 ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga acacccccat
     1681 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag
     1741 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1801 gatcactctc ggcatggacg agctgtacaa gtgacaactt tattatacaa agttggcatt
     1861 ataaaaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa
     1921 atcattattt ggagctccat ggtagcgtta acgcggccgc gatatcccct atagtgagtc
     1981 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2041 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     2101 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2161 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2221 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2281 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2341 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2401 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2461 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     2521 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     2581 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     2641 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     2701 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     2761 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     2821 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     2881 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     2941 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3001 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3061 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3121 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3181 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3241 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3301 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3361 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3421 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3481 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3541 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3601 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3661 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3721 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3781 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3841 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3901 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     3961 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     4021 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     4081 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     4141 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     4201 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
     4261 ttgtaaaacg acggccagtc ttaagctcgg gccctgcagc tctagagctc gaattctaca
     4321 ggtcactaat accatctaag tagttggttc atagtgactg catatgttgt gttttacagt
     4381 attatgtagt ctgtttttta tgcaaaatct aatttaatat attgatattt atatcatttt
     4441 acgtttctcg ttcaactttc ttgtacaaag tggc
//
