LOCUS       sequence-243654-        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"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             786..827
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             834..1790
                     /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             1797..1823
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             join(1863..1865,1866..1868,1869..2582)
                     /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(2169..2188)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2516..2537
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2583..2678)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2715..2734)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2716..2734)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2739..2755)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2739..2755)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2857..2876)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2868..3677
                     /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(2935..2954)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3824..4412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4313..4332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggctg ggcgcgccgg tggaggcggt tcaggaggtg
      781 gcatgggcaa gcccatcccc aaccccctgc tgggcctgga cagcaccgct agcaaagaca
      841 atactgtgcc tctgaagctg atcgctctcc tggctaatgg cgagttccat agtggcgaac
      901 agctgggaga aaccctgggc atgtccaggg ccgctatcaa caagcacatt cagactctgc
      961 gcgactgggg cgtggacgtg ttcaccgtgc ccggaaaggg ctactctctg cccgagccta
     1021 tcccgctgct gaacgctaaa cagattctgg gacagctgga cggcgggagc gtggcagtcc
     1081 tgcctgtggt cgactccacc aatcagtacc tgctggatcg aatcggcgag ctgaagagtg
     1141 gggatgcttg cattgcagaa tatcagcagg cagggagagg aagcagaggg aggaaatggt
     1201 tctctccttt tggagctaac ctgtacctga gtatgttttg gcgcctgaag cggggaccag
     1261 cagcaatcgg cctgggcccg gtcatcggaa ttgtcatggc agaagcgctg cgaaagctgg
     1321 gagcagacaa ggtgcgagtc aaatggccca atgacctgta tctgcaggat agaaagctgg
     1381 caggcatcct ggtggagctg gccggaataa caggcgatgc tgcacagatc gtcattggcg
     1441 ccgggattaa cgtggctatg aggcgcgtgg aggaaagcgt ggtcaatcag ggctggatca
     1501 cactgcagga agcagggatt aacctggaca ggaatactct ggccgctacg ctgatccgag
     1561 agctgcgggc agccctggaa ctgttcgagc aggaaggcct ggctccatat ctgccacggt
     1621 gggagaagct ggataacttc atcaatagac ccgtgaagct gatcattggg gacaaagaga
     1681 ttttcgggat tagccggggg attgataaac agggagccct gctgctggaa caggacggag
     1741 ttatcaaacc ctggatgggc ggagaaatca gtctgcggtc tgccgaaaag ctgcagctgc
     1801 ctcccctgga gcgcctgacc ctggacggag gcggtggaag cggcggaggt gggcgcgcct
     1861 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1921 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1981 acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca
     2041 ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
     2101 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2161 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2221 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2281 acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga
     2341 acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg
     2401 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2461 actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     2521 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     2581 gacaacttta ttatacaaag ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa
     2641 cgaacaggtc actatcagtc aaaataaaat cattatttgg agctccatgg tagcgttaac
     2701 gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     2761 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     2821 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     2881 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     2941 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     3001 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     3061 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     3121 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     3181 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     3241 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     3301 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     3361 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     3421 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     3481 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     3541 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     3601 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     3661 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     3721 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     3781 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3841 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3901 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3961 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4021 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4081 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4141 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4201 acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4261 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4321 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4381 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4441 tcctggcctt ttgctggcct tttgctcaca tgtt
//
