LOCUS       sequence-246338-        4438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for making C-terminal TurboID-mVenus fusion 
            with nuclear and non-nuclear proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4438)
  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 4438)
  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-mVenus-STOP-L3
FEATURES             Location/Qualifiers
     source          1..4438
                     /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             join(1079..1081,1082..1084,1085..1798)
                     /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(1385..1404)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1732..1753
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1799..1894)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1931..1950)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1932..1950)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1955..1971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1955..1971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2073..2092)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2084..2893
                     /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(2151..2170)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3040..3628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3529..3548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3782..3799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3958..3985)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4077..4163)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4212..4234
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4226..4243
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4227..4243
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4314..4437)
                     /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 agggaggcgg tggaagcggc ggaggtgggc gcgcctccat
     1081 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1141 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1201 caagctgacc ctgaagctga tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1261 cgtgaccacc ctgggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca
     1321 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1381 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1441 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1501 gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     1561 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccga
     1621 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1681 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1741 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtgaca
     1801 actttattat acaaagttgg cattataaaa aagcattgct tatcaatttg ttgcaacgaa
     1861 caggtcacta tcagtcaaaa taaaatcatt atttggagct ccatggtagc gttaacgcgg
     1921 ccgcgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     1981 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     2041 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     2101 ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     2161 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg ggaagcccga
     2221 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     2281 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     2341 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa cagcattcca
     2401 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     2461 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     2521 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     2581 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     2641 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     2701 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     2761 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     2821 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     2881 tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca ttacgctgac
     2941 ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3001 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3061 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3181 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3781 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     3841 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     3901 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     3961 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     4021 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     4081 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     4141 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     4201 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggccctg
     4261 cagctctaga gctcgaattc tacaggtcac taataccatc taagtagttg gttcatagtg
     4321 actgcatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
     4381 atatattgat atttatatca ttttacgttt ctcgttcaac tttcttgtac aaagtggc
//
