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pMK107

Expression clone for an auxin-based degron (AID) system, CENPH-aid.

Catalog number RDB08470
Resource name pMK107
Alternative name OsTIR1-9Myc::IRES::CENPH-aid (kan/neoR)
Clone info. Expression clone for an auxin-based degron (AID) system, CENPH-aid. Refer Supplementary figure 11 (CENPH-aid) of Nat. Methods, 6, 917-922, 2009; PMID: 19915560.
Although this clone is constructed on the pIRES-EGFP, it doesn't contain GFP. Accession numbers, AtIAA17 (GeneID 839568), CENPH (Gallus gallus). To make 0.5 M stock solution, dissolve indole-3-acetic acid (IAA) (Sigma-Aldrich, 45533) or 1-naphthaleneacetic acid (NAA) (Sigma-Aldrich, 35745) in 100 % DMSO. IAA solution can be made in 100 % ethanol. Aliquots can be kept for 6 months in the dark at -20oC (Devrekanli, A., Kanemaki, M.T. Methods in Molecular Biology, 1369, 257-278, 2016).
Vector backbone pIRES-EGFP (Plasmid)
Selectable markers Kan^r
Gene/insert name Gallus gallus CENPH cDNA Oryza sativa OsTIR1 Arabidopsis thaliana AtIAA17
Depositor|Developer Kanemaki, Masato |
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External Database
Gallus gallus CENPH Oryza sativa OsTIR1 Arabidopsis thaliana AtIAA17

      Reference sequence
        

      Distribution information

      Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
      Terms and conditions set forth by the DEPOSITOR In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature(s) designated by the DEPOSITOR is requested (Nishimura, K., Nat. Methods., 6, 917-922 (2009)). This DNA clone is used for academic research purpose only. This is not to be used on a commercial basis. For use of this DNA clone except for internal academic research, the RECIPIENT must contact a NIG license representative at chizai@nig.ac.jp.
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      RDB08470 pMK107 DNA solution

      Ordering Information [in Japanese] [in English]
      Please review the QC test results indicated by check icon below as well as clone information before placing your order.

      References and tips

      Electronic file

      Sequence RDB08470z.seq
      Remarks, protocol and/or map (pdf) RDB08470.pdf

      Original reference

      original Nishimura, K., An auxin-based degron system for the rapid depletion of proteins in nonplant cells. Nat Methods. 6(12), 917-22, (2009). PMID 19915560.

      Further references such as user reports and related articles (go to bottom)

      Featured content

      Featured content Auxin Inducible Degron (AID) System (English text)
      Featured content Auxin Inducible Degron (AID) System (Japanese text)

      Sequence information

      RIKEN BRC has sequenced portions of this material for quality test.
      Please review the QC test results indicated by check icon as well as clone information before placing your order.

      Test sheet RDB08470_A5C2p1.pdf check

      Nucleotide sequence of a portion of this resource (if available).

      Primer: CMV-Forward
      Sequence file: RDB08470_A5C2a.seq check
      >08470_08470_A5C2_CMV-Forward_A08_02.ab1
          1 CGTCACTCCG CCCATTGACG CAATGGGCGG TAGGCGTGTA CGGTGGGAGG TCTATATAAG
         61 CAGAGCTGGT TTAGTGAACC GTCAGATCCG CTAGCGCTAC CGGACTCAGA TCTCGAGATG
        121 ACGTACTTCC CGGAGGAGGT GGTGGAGCAC ATCTTCAGCT TCCTGCCGGC GCAGCGCGAC
        181 CGCAACACGG TCTCGCTCGT CTGCAAGGTG TGGTACGAGA TCGAGAGGCT GAGCCGCCGC
        241 GGCGTCTTCG TGGGCAACTG CTACGCCGTG CGCGCCGGCC GCGTCGCCGC GCGGTTCCCC
        301 AACGTGCGGG CGCTCACGGT GAAGGGGAAG CCCCACTTCG CCGACTTCAA CCTCGTGCCC
        361 CCCGACTGGG GCGGCTACGC GGGGCCGTGG ATCGAGGCGG CCGCGAGGGG ATGCCACGGC
        421 CTGGAGGAGC TCAGGATGAA GCGGATGGTG GTGTCCGACG AGAGCCTCGA GCTGCTGGCT
        481 CGCTCGTTCC CGCGGTTCAG GGCTCTTGTT CTTATCAGCT GCGAGGGGTT CAGCACTGAC
        541 GGGCTAGCCG CCGTCGCGAG CCATTGCAAG CTTCTGAGGG AGTTGGATTT GCAGGAAAAT
        601 GAAGTGGAGG ATCGAGGGCC TAGGTGGCTT TCCTGCTTCC CTGATTCCTG CACATCACTT
        661 GTCTCATTGA ATTTTGCCTG CATCAAAGGG GAGGTTAATG CTGGTTCACT GGAGAGACTT
        721 GTTAGCAGGT CCCCAAACCT GCGGAGTTTG AGGCTGAATC GATCTGTATC GGTAGATACA
        781 CTTGCAAAGA TACTACTGCG TACCCCTAAC TTGGAGGATT TGGGGACAGG GAATTTGACA
        841 GATGACTTCC AAACTGAGTC CTACTTTAAG CTTACCAGTG CTCTGGAGAA ATGCAAGATG
        901 TTGAGGAGTT TGTCTGGATT CTGGGATGCT TCTCCTGTTT GCCTGTCATT TATCTACCCC
        961 CCTGTGTGCT CAACTGACAG GATTGAACTT GAGCTATGCA CCCACACTTG GATGCTTTCT
       1021 GACCCTTACA AAAATGATTT AGCCGCTGTG TGAAGCTCCT ACCGACGTTT GGGTACTGCA
       1081 TTGTAATCTC GACCAACGCC TGCCAGTTGT TGACCTCCAG CTTGCAAAGA ACCTTGGCCA
      1141 G
      //
      Primer: ME-1290Rv
      Sequence file: RDB08470_A5C2b.seq check
      >08470_08470_A5C2_ME-1290Rv_B08_05.ab1
          1 AACGCATTTC ATTTTTATGT TTCAGGTTCA GGGGGAGGTG TGGGAGGTTT TTTAAAGCAA
         61 GTAAAACCTC TACAAATGTG GTATGGCTGA TTATGATCTA GAGTCGCGGC CGCTCAAGCT
        121 CTGCTCTTGC ACTTCTCCAT CGCCCTCGGA GCGAGACCAA TGGCATCCGA TCCTTTCATG
        181 AGACGTAAAC GCTTGCATGT ATCGACGAAC ATTGGCCAAG GAACGTCGCC GACGAGCATC
        241 CAATCACCGT CTTTGTCTTC ATAAGAGGGA ACATAGTCCC AGCTATTCAC CAAATCCATC
        301 AATTTCCTCT CATTCATGAA GTCTATCATT CCTTCTTCTC CTCCATGTTT GCCCATGGTA
        361 AAAGAGCTGA ACATGTTGGA CAAAGCATTA GAAAGCTCAT CGTAGCTTTT ATACATCCTC
        421 AAATCGATTT TCCTCAAGTA CGGTGCTCCG TCCATTGATA CCTTCACGAA CGCCGCCGCC
        481 TCCGGGCCAC CGCTTGATTT TTGGCAGGAA ACCATCACGT TCTTCCGGTA TGATCTCACC
        541 GGTGGCCATC CCACAACTTG TGCCTTGGCC GGAGGTTTGG CTGGATCTTT AGGACAAGCA
        601 CTCTTCTCCT TGGAATCAAA AGTCACGACG TCATGAGTCG TAGATCCTTC CTTGTTTGCA
        661 GGCTCATTAT TCAGATTTAG CTTCAGATCA ACCGTCTCTG AGAACCCTCT CTTGTTTCCG
        721 GTTACCGGAG CCACTGTATC TCCACCGGGA AGACCAAGAC ACAGCTCAGT CTCCCTCAGA
        781 TTCAGCTCGA CACTGCCCAT GGTTGTGGCG GCACCCGCTC CAGCGCCTGC ACCAGCTCCA
        841 GTACTCAAGA GATTCTGAAT GGAGATATCT TTCTCGAGCT GTAGAATAAT TGCTCTTAAA
        901 GATGGATCTT TCGCCCAGTT GATTTGGCAT CCGACTATGA TATTCTGGAG TATATTTTGA
        961 ACTAACGTAG TCACCTTCCC TTTCATACTG TACATTCCTA TGGGATAAAC TTTCAACTTT
       1021 TCAGCAATTT TCATTCTGAC TACTTCTTTT TTTTTTTCTC ACTGTGTAAA AACTCGCAAG
       1081 CGCCTTCTTC TCCGTTCCGG ACTGTCAGAG TATCTTTCAT CTATGTCAAT TCCAAATTTC
      1141 CTCTCTCTCT A
      //
      Primer: IRES-R
      Sequence file: RDB08470_A5C2c.seq check
      >08470_08470_A5C2_IRES-R_C08_08.ab1
          1 GCATAGCGGC TTCGGCAGTA ACGTTAGGGG GGGGGGAGGG AGAGGGGCGG ATCCCGGGGA
         61 TCTCTTGAAT GATCGTTCCA CTTTTTAGCT AGTGGATCCG TTCAAGTCTT CTTCTGAGAT
        121 TAATTTTTGT TCACCGTTCA AGTCTTCCTC GGAGATTAGC TTTTGTTCAC CGTTCAAATC
        181 TTCTTCAGAA ATCAACTTTT GTTCACCTCT AGAGGATCCG TTCAAGTCTT CTTCTGAGAT
        241 TAATTTTTGT TCACCGTTCA AGTCTTCCTC GGAGATTAGC TTTTGTTCAC CGTTCAAATC
        301 TTCTTCAGAA ATCAACTTTT GTTCACCTCT AGAGGATCCG TTCAAGTCTT CTTCTGAGAT
        361 TAATTTTTGT TCACCGTTCA AGTCTTCCTC GGAGATTAGC TTTTGTTCAC CGTTCAAATC
        421 TTCTTCAGAA ATCAACTTTT GTTCACCACT AGCAGCAGAA CCGGAGTCGA CTAGGATTTT
        481 AACAAAATTT GGTGCATCAT CCCTCGCCCC AGCAGTTGTG CGGTACACAT ATAATTTCTC
        541 CACTTTAGGC AGATCTCCAT GGTTTTCCTC CATTTCATTG CTACCATCCC GCTCATTTAT
        601 GACCTCAACA TTGAGCATCG GCATCTTTGA CGCAAGGACT TGGCACCCCT TTAACGTGAC
        661 ATTGCAAGAT GACATCCAAA GGGATCGCAT TGTCTCGTAC CTAGCAAAAT TCCCCAAGAG
        721 TGCAGCATCA CCAAACGGGC TATCTCTTAT CTCCAGTTTC CTTAAATTCT TGCATCCATT
        781 CATAACATGC ATCATACCCT TATCACTGTC ACCAGCAAAT GCTATAGAAA GCATCTCAAG
        841 TTGTTTTGCA TATTTCCCAA TATACATGAA AACTTTGTCT GTGAGAAGAC CAGATATTGA
        901 CAAACGTTGT AATCCCTTGC ACTCACGAAC AATAGCTCCA AAGCCTTCAT CTAATGGTTG
        961 GCTTGTCACA ACATCAGGCT TCCCTGGCTC AAGAATACAA AGTCTGAATC GTGTGAAATT
       1021 TGGGACAGTT CTTGGCGACA GTAACTAGTG CAGCATTTAG TCATTTGGGT GACAGAGTAC
       1081 AGTAGTGAGT TCAGTTTTGC ACAGGCCAGA TACCTGCACA GTTCCCTTCC TCTGTCACTG
      1141 CAGAAAATTA ACCCAAG
      //
      Primer: IRES-F
      Sequence file: RDB08470_A5C2d.seq check
      >08470_08470_A5C2_IRES-F_D08_11.ab1
          1 CGGTCATTCT GTGTTTAGTC GAGGTTAAAA AACGTCTAGG CCCCCCGAAC CACGGGGACG
         61 TGGTTTTCCT TTGAAAAACA CGATGATAAT ATGGCCACAA CCATGTGCGA TATCGAATTC
        121 ACCATGGCGG GGCGGCTGTC GGAGTCCGTC GGGTCCGGGC CGGGCGCTGA GGCCGAGACG
        181 GCGGCCGATC CGGACGCGAA GCGGGACGTC CTGGAGCACC TCTGCGCGAG GACGCATCTG
        241 AAGCAGCTGG TGATGGAGTT CGACACCGCG TGCCCCCCCG ATGAAGGGTG CAACAGTGGC
        301 GCGGAAGTAA ACTTTATCGA AAGTGCCAAA GAAACTTTGG AAGAAGTGGG GAAAGTAAAA
        361 AGTGCCTTTG AAAGCAAAGC ATTAGTCATA AAAAGGATCC AGCTTATGGA TGCTCTGAGA
        421 AAGAGAGTGA AAGAAAATGA TGGTTGCGCA CGACTGATCG TGGAAACAAT GAGAGACATA
        481 ATCAAGCTCA ACTGGGAAAT AATTCAAGCT CATCAGCAAG CACGTGTGAT TAGAGAGAAT
        541 TTGAATGACA TTAGAAGGAA AAGATACTTT CTGAAACAGG CCGAAGGAGA GAAGGCGCTG
        601 CGAATTTTTA CCACAGTGAG GAAAAAAAAG GAAGTAGTCA GAATGAAAAT TGCTGAAAAG
        661 TTGAAGTTTA TCCATAGGAA TGTACAGTAT GAAAGGAAGG TGACTACGTT AGTTCAAAAT
        721 ATACTCCAGA ATATCATAGT CGGATGCCAA ATCAACTGGG CGAAAGATCC ATCTTTAAGA
        781 GCAATTATTC TACAGCTCGA GAAAGATATC TCCATTCAGA ATCTCTTGAG TACTGGAGCT
        841 GGTGCAGGCG CTGGAGCGGG TGCCGCCACA ACCATGGGCA GTGTCGAGCT GAATCTGAGG
        901 GAGACTGAGC TGTGTCTTGG TCTTCCCGGT GGAGATACAG TGGCTCCGGT AACCGGAAAC
        961 AAGAGAGGGT TCTCAGAGAC GGTTGATCTG AAGCTAAATC TGAATAAATG AGCCTGCAAA
       1021 CAAGGAAAGG ATCTACGACT CATGACGTCG TGACTTTTTG ATTTCGACGA GAGAGTGCTG
       1081 TCCTAAAGAT TCAGCGAACT CGGACAGCAC AGCTGTGGGA ATGGCACGTG GAATCATACT
      1141 GGCAGCAACC GAG
      //
      Primer: SV40pro_F_V2
      Sequence file: RDB08470_A5C2e.seq check
      >08470_08470_A5C2_SV40pro_F_V2_E08_14.ab1
          1 ACCATGGCAC TCTTTTGAGG CTAGGCTTTT GCAAGATCGA TCAAGAGCAC ACGGATGAGG
         61 ATCGTTTCGC ATGATTGAAC AAGATGGATT GCACGCAGGT TCTCCGGCCG CTTGGGTGGA
        121 GAGGCTATTC GGCTATGACT GGGCACAACA GACAATCGGC TGCTCTGATG CCGCCGTGTT
        181 CCGGCTGTCA GCGCAGGGGC GCCCGGTTCT TTTTGTCAAG ACCGACCTGT CCGGTGCCCT
        241 GAATGAACTG CAAGACGAGG CAGCGCGGCT ATCGTGGCTG GCCACGACGG GCGTTCCTTG
        301 CGCAGCTGTG CTCGACGTTG TCACTGAAGC GGGAAGGGAC TGGCTGCTAT TGGGCGAAGT
        361 GCCGGGGCAG GATCTCCTGT CATCTCACCT TGCTCCTGCC GAGAAAGTAT CCATCATGGC
        421 TGATGCAATG CGGCGGCTGC ATACGCTTGA TCCGGCTACC TGCCCATTCG ACCACCAAGC
        481 GAAACATCGC ATCGAGCGAG CACGTACTCG GATGGAAGCC GGTCTTGTCG ATCAGGATGA
        541 TCTGGACGAA GAGCATCAGG GGCTCGCGCC AGCCGAACTG TTCGCCAGGC TCAAGGCGAG
        601 CATGCCCGAC GGCGAGGATC TCGTCGTGAC CCATGGCGAT GCCTGCTTGC CGAATATCAT
        661 GGTGGAAAAT GGCCGCTTTT CTGGATTCAT CGACTGTGGC CGGCTGGGTG TGGCGGACCG
        721 CTATCAGGAC ATAGCGTTGG CTACCCGTGA TATTGCTGAA GAGCTTGGCG GCGAATGGGC
        781 TGACCGCTTC CTCGTGCTTT ACGGTATCGC CGCTCCCGAT TCGCAGCGCA TCGCCTTCTA
        841 TCGCCTTCTT GACGAGTTCT TCTGAGCGGG ACTCTGGGGT TCGAAATGAC CGACCAAGCG
        901 ACGCCCAACC TGCCATCACG AGATTTCGAT TCCACCGCCG CCTTCTATGA AAGGTTGGGC
        961 TTCGGAATCG TTTTCCGGGA CGCCGGCTGG ATGATCCTCC AGCGCGGGGA TCTCATGCTG
       1021 GAGTTCTTCG CCACCCTAGG GGGAGGCTAA CTGAACACGG AAGAGACATA CCGGAGCACC
       1081 CGCCGCTTAT GACGGCAATT AAAAGACCGG AATAAAACGC CACGGTGTTG GGATCGGATT
      1141 TGAATCTA
      //

      Please visit Sequencing and PCR primers for primer information.


      References

      Original, user report and related articles

      original Nishimura, K., An auxin-based degron system for the rapid depletion of proteins in nonplant cells. Nat Methods. 6(12), 917-22, (2009). PMID 19915560.
      reference Devrekanli, A., Conditional Budding Yeast Mutants with Temperature-Sensitive and Auxin-Inducible Degrons for Screening of Suppressor Genes. Methods in Molecular Biology 1369: 257-278 (2016). PMID 26519318.
      reference Nishimura, K., Rapid Depletion of Budding Yeast Proteins via the Fusion of an Auxin-Inducible Degron (AID). Curr. Protoc. Cell Biol. \64: 20.9.1-16 (2014). PMID 25181302.
      reference Holland, A.J., Inducible, reversible system for the rapid and complete degradation of proteins in mammalian cells. Proc. Natl. Acad. Sci. U.S.A. 109 (49): E3350-7 PMID 23150568.

      2021.11.02

      GNP_filter3_RDBDEP_html_211029.pl