LOCUS       sequence-243656-        4288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gateway entry vector for making C-terminal miniTurbo-mVenus fusion 
            with non-nuclear proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4288)
  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 4288)
  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-miniTurbo-NES-mVenus-STOP-L3
FEATURES             Location/Qualifiers
     source          1..4288
                     /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..1604
                     /codon_start=1
                     /product="N-terminally truncated mutant of the E. coli 
                     biotin ligase BioID engineered for improved catalytic 
                     efficiency (Branon et al., 2018)"
                     /label=miniTurbo
                     /translation="IPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSGDACI
                     AEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKLGADK
                     VRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGWITLQ
                     EAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGDKEIF
                     GISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     CDS             1611..1637
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             join(1677..1679,1680..1682,1683..2396)
                     /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(1983..2002)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2330..2351
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2397..2492)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2529..2548)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2530..2548)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2553..2569)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2553..2569)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2671..2690)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2682..3491
                     /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(2749..2768)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3638..4226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4127..4146
                     /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 agcatcccgc
      841 tgctgaacgc taaacagatt ctgggacagc tggacggcgg gagcgtggca gtcctgcctg
      901 tggtcgactc caccaatcag tacctgctgg atcgaatcgg cgagctgaag agtggggatg
      961 cttgcattgc agaatatcag caggcaggga gaggaagcag agggaggaaa tggttctctc
     1021 cttttggagc taacctgtac ctgagtatgt tttggcgcct gaagcgggga ccagcagcaa
     1081 tcggcctggg cccggtcatc ggaattgtca tggcagaagc gctgcgaaag ctgggagcag
     1141 acaaggtgcg agtcaaatgg cccaatgacc tgtatctgca ggatagaaag ctggcaggca
     1201 tcctggtgga gctggccgga ataacaggcg atgctgcaca gatcgtcatt ggcgccggga
     1261 ttaacgtggc tatgaggcgc gtggaggaaa gcgtggtcaa tcagggctgg atcacactgc
     1321 aggaagcagg gattaacctg gacaggaata ctctggccgc tatgctgatc cgagagctgc
     1381 gggcagccct ggaactgttc gagcaggaag gcctggctcc atatctgtca cggtgggaga
     1441 agctggataa cttcatcaat agacccgtga agctgatcat tggggacaaa gagattttcg
     1501 ggattagccg ggggattgat aaacagggag ccctgctgct ggaacaggac ggagttatca
     1561 aaccctggat gggcggagaa atcagtctgc ggtctgccga aaagctgcag ctgcctcccc
     1621 tggagcgcct gaccctggac ggaggcggtg gaagcggcgg aggtgggcgc gcctccatgg
     1681 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1741 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1801 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1861 tgaccaccct gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc
     1921 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1981 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     2041 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     2101 tggagtacaa ctacaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca
     2161 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
     2221 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2281 tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2341 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtgacaac
     2401 tttattatac aaagttggca ttataaaaaa gcattgctta tcaatttgtt gcaacgaaca
     2461 ggtcactatc agtcaaaata aaatcattat ttggagctcc atggtagcgt taacgcggcc
     2521 gcgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     2581 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa
     2641 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg
     2701 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
     2761 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg
     2821 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
     2881 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
     2941 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg
     3001 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
     3061 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
     3121 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
     3181 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct
     3241 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
     3301 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
     3361 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
     3421 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
     3481 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt
     3541 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     3601 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3661 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3721 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3781 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3841 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3901 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3961 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4021 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4081 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4141 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4201 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4261 ccttttgctg gccttttgct cacatgtt
//
