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pMK106

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

Catalog number RDB08469
Resource name pMK106
Alternative name OsTIR1-9Myc::IRES::aid-CENPH (kan/neoR)
Clone info. Expression clone for an auxin-based degron (AID) system, aid-CENPH. Refer Supplementary figure 11 (aid-CENPH) 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, OsTIR1 (GeneID 4335696); 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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      RDB08469 pMK106 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 RDB08469z.seq
      Remarks, protocol and/or map (pdf) RDB08469.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 RDB18378_B0Hhp1-1.pdf check

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

      Primer: CMV_Forward
      Region: CMV pro,OsTIR1 5'
      Sequence file: RDB18378_B0Hha.seq check
      >08469_18378_B0Hh_3_CMV_Forward_A11_02_ABI24.ab1
          1 NNNNNNNNNN NNNNNNTNNN GCNNATGGGC GGTANGCGTG TACGGTGGGN NGTCTATATA
         61 AGCAGAGCTG GTTTAGTGAA CCGTCAGATC CGCTAGCGCT ACCGGACTCA GATCTCGAGA
        121 TGACGTACTT CCCGGAGGAG GTGGTGGAGC ACATCTTCAG CTTCCTGCCG GCGCAGCGCG
        181 ACCGCAACAC GGTCTCGCTC GTCTGCAAGG TGTGGTACGA GATCGAGAGG CTGAGCCGCC
        241 GCGGCGTCTT CGTGGGCAAC TGCTACGCCG TGCGCGCCGG CCGCGTCGCC GCGCGGTTCC
        301 CCAACGTGCG GGCGCTCACG GTGAAGGGGA AGCCCCACTT CGCCGACTTC AACCTCGTGC
        361 CCCCCGACTG GGGCGGCTAC GCGGGGCCGT GGATCGAGGC GGCCGCGAGG GGATGCCACG
        421 GCCTGGAGGA GCTCAGGATG AAGCGGATGG TGGTGTCCGA CGAGAGCCTC GAGCTGCTGG
        481 CTCGCTCGTT CCCGCGGTTC AGGGCTCTTG TTCTTATCAG CTGCGAGGGG TTCAGCACTG
        541 ACGGGCTAGC CGCCGTCGCG AGCCATTGCA AGCTTCTGAG GGAGTTGGAT TTGCAGGAAA
        601 ATGAAGTGGA GGATCGAGGG CCTAGGTGGC TTTCCTGCTT CCCTGATTCC TGCACATCAC
        661 TTGTCTCATT GAATTTTGCC TGCATCAAAG GGGANGTTAA TGCTGGTTCA CTGGAGAGAC
        721 TTGTTAGCAG GTCCNCAAAC CTGCGGANTT TGANGCTGAN TCGATCTGTA TCGGTAGATA
        781 CACTTGCNAA GATACTACTG CGTACNNCTA ACTTGNNNGA NTTGGGGANA NGGAATTTGN
        841 CAGATGANTT CCNAACTGAN TCCTANTTTN AGCTTACCAN NNCTCTGGAN AANTGNNANA
      901 NGTTGANGAN TTTGTCNGGN NTCTGGNNNN NNTCNNCNNG TTNNNNNNNN NN
      //
      Primer: IRES-R
      Region: 9XMyc,OaTIR1 3'
      Sequence file: RDB18378_B0Hhb.seq check
      >08469_18378_B0Hh_3_IRES-R_C11_08_ABI24.ab1
          1 NNNNNNNNNN NNNNNNNNNN NNNNNNGTTA NGGGGGGGGG NNGGNNNNGG GCGGNTCCCG
         61 GGGNNCTCTT GAATGATCGT TCCACTTTTT AGCTAGTGGA TCCGTTCAAG TCTTCTTCTG
        121 AGATTAATTT TTGTTCACCG TTCAAGTCTT CCTCGGAGAT TAGCTTTTGT TCACCGTTCA
        181 AATCTTCTTC AGAAATCAAC TTTTGTTCAC CTCTAGAGGA TCCGTTCAAG TCTTCTTCTG
        241 AGATTAATTT TTGTTCACCG TTCAAGTCTT CCTCGGAGAT TAGCTTTTGT TCACCGTTCA
        301 AATCTTCTTC AGAAATCAAC TTTTGTTCAC CTCTAGAGGA TCCGTTCAAG TCTTCTTCTG
        361 AGATTAATTT TTGTTCACCG TTCAAGTCTT CCTCGGAGAT TAGCTTTTGT TCACCGTTCA
        421 AATCTTCTTC AGAAATCAAC TTTTGTTCAC CACTAGCAGC AGAACCGGAG TCGACTAGGA
        481 TTTTAACAAA ATTTGGTGCA TCATCCCTCG CCCCAGCAGT TGTGCGGTAC ACATATAATT
        541 TCTCCACTTT AGGCAGATCT CCATGGTTTT CCTCCATTTC ATTGCTACCA TCCCGCTCAT
        601 TTATGACCTC AACATTGAGC ATCGGCATCT TTGACGCAAG GACTTGGCAC CCCTTTAACG
        661 TGACATTGCA AGATGACATC CAAAGGGATC GCATTGTCTC GTACCTAGCA AAATTCCCCA
        721 AGAGTGCAGC ATCACCAAAC GGGCTATCTC TTATCTCCNG TTTCCTTAAA TTCTTGCATC
        781 CATTCATAAC ATGCATCATA CCCTTATCAC TGTCACCAGC AAATGCTATA NAAAGCATCT
        841 CNAGTTGTTT TGCATATTTC CNAATATACA TGAAAACTTT GTCTGTGAGA NNANCAGATA
        901 TTGANAAACG TTNNNATCCC NNGCACNCNC NAACNNNANN NNCNAANNNT TNNTCNNATG
        961 GTNNNNTTGN CNCNNNNTCN GNNTNNCNNG GNNNNNNNAN NNNNANNNCN GNANNNNNNN
      1021 NNN
      //
      Primer: IRES-F
      Region: IRES2,AID 5'
      Sequence file: RDB18378_B0Hhc.seq check
      >08469_18378_B0Hh_3_IRES-F_D11_11_ABI24.ab1
          1 NNNNNNNNNN NNNNTNNNTC GNNGTTNNNA AACGTCTAGG CCCCCCGAAC CNCGGGGNCG
         61 TGGTTTTCCT TTGAAAAACA CGATGATAAT ATGGCCACAA CCATGGGCAG TGTCGAGCTG
        121 AATCTGAGGG AGACTGAGCT GTGTCTTGGT CTTCCCGGTG GAGATACAGT GGCTCCGGTA
        181 ACCGGAAACA AGAGAGGGTT CTCAGAGACG GTTGATCTGA AGCTAAATCT GAATAATGAG
        241 CCTGCAAACA AGGAAGGATC TACGACTCAT GACGTCGTGA CTTTTGATTC CAAGGAGAAG
        301 AGTGCTTGTC CTAAAGATCC AGCCAAACCT CCGGCCAAGG CACAAGTTGT GGGATGGCCA
        361 CCGGTGAGAT CATACCGGAA GAACGTGATG GTTTCCTGCC AAAAATCAAG CGGTGGCCCG
        421 GAGGCGGCGG CGTTCGTGAA GGTATCAATG GACGGAGCAC CGTACTTGAG GAAAATCGAT
        481 TTGAGGATGT ATAAAAGCTA CGATGAGCTT TCTAATGCTT TGTCCAACAT GTTCAGCTCT
        541 TTTACCATGG GCAAACATGG AGGAGAAGAA GGAATGATAG ACTTCATGAA TGAGAGGAAA
        601 TTGATGGATT TGGTGAATAG CTGGGACTAT GTTCCCTCTT ATGAAGACAA AGACGGTGAT
        661 TGGATGCTCG TCGGCGACGT TCCTTGGCCA ATGTTCGTCG ATACATGCAA GCGTTTACGT
        721 CTCATGAAAG GATCGGATGC CATTGGTCTC GCTCCGAGGG CGATGGAGAA GTGCAAGANC
        781 AGANCTGCGG CCGGANCTGG TGCANGCGCT GGANCGGGTG CCGATATCGA ATTCACCANG
        841 GCGGGGCGGC TGTCGGANNN CGTCNGGNNC NGGCCGGGCG CTNANGCCNA NACNGCGGCC
      901 NATNCNNNNG CNAANCGGGN NNNNNNNNN
      //
      Primer: SV40polyA_R2
      Region: bGHpA,chiken CENP-H 3'
      Sequence file: RDB18378_B0Hhd.seq check
      >08469_18378_B0Hh_3_SV40polyA_R2_B11_05_ABI24.ab1
          1 NNNNNNNNCN NNNNNNNNNN NGTTNNNAAC AACAATTGCA TTCNTTTTAT GTTTCAGGTT
         61 CAGGGGGAGG TGTGGGAGGT TTTTTAAAGC AAGTAAAACC TCTACAAATG TGGTATGGCT
        121 GATTATGATC TAGAGTCGCG GCCGCAGTGA TCACAGTACT CTACAAGAGA TTCTGAATGG
        181 AGATATCTTT CTCGAGCTGT AGAATAATTG CTCTTAAAGA TGGATCTTTC GCCCAGTTGA
        241 TTTGGCATCC GACTATGATA TTCTGGAGTA TATTTTGAAC TAACGTAGTC ACCTTCCTTT
        301 CATACTGTAC ATTCCTATGG ATAAACTTCA ACTTTTCAGC AATTTTCATT CTGACTACTT
        361 CCTTTTTTTT CCTCACTGTG GTAAAAATTC GCAGCGCCTT CTCTCCTTCG GCCTGTTTCA
        421 GAAAGTATCT TTTCCTTCTA ATGTCATTCA AATTCTCTCT AATCACACGT GCTTGCTGAT
        481 GAGCTTGAAT TATTTCCCAG TTGAGCTTGA TTATGTCTCT CATTGTTTCC ACGATCAGTC
        541 GTGCGCAACC ATCATTTTCT TTCACTCTCT TTCTCAGAGC ATCCATAAGC TGGATCCTTT
        601 TTATGACTAA TGCTTTGCTT TCAAAGGCAC TTTTTACTTT CCCCACTTCT TCCAAAGTTT
        661 CTTTGGCACT TTCGATAAAG TTTACTTCCG CGCCACTGTT GCACCCTTCA TCGGGGGGGC
        721 ACGCGGTGTC GAACTCCATC ACCAGCTGCT TCAGATGCGT CCTCGCGCAN ANGTGCTCCA
        781 GGACGTCCCG CTTCGCGTCC GGNTCGGCCG CCGTCTCGGC CTCAGCGCCC GGCCCGGANC
        841 CNANGGNNTC CGACAGCCGC CCCGCCANGG NGAANTNNAT ATCGGCACCN GCNCCAGCGC
        901 NNGNNCCANC TCCNGNCGCA NCNNNGCNNN NGCNNTTNNN CNTCNNCCTC NNNNCNNNNN
      961 NNNNGNNNNN NNNNN
      //
      Primer: CENP-M_R
      Region: chiken CENP-H 5',AID 3'
      Sequence file: RDB18378_B0Hhe.seq check
      >08469_18378_B0Hh_3_CENP-M_R_E11_14_ABI24.ab1
          1 NNNNNNNNNN NNNNNNNNNN NNNNTCNCNN GCTGCTTNNG ATGCGTCCTC GCGCAGANGT
         61 GCTCCAGGAC GTCCCGCTTC GCGTCCGGAT CGGCCGCCGT CTCGGCCTCA GCGCCCGGCC
        121 CGGACCCGAC GGACTCCGAC AGCCGCCCCG CCATGGTGAA TTCGATATCG GCACCCGCTC
        181 CAGCGCCTGC ACCAGCTCCG GCCGCAGCTC TGCTCTTGCA CTTCTCCATC GCCCTCGGAG
        241 CGAGACCAAT GGCATCCGAT CCTTTCATGA GACGTAAACG CTTGCATGTA TCGACGAACA
        301 TTGGCCAAGG AACGTCGCCG ACGAGCATCC AATCACCGTC TTTGTCTTCA TAAGAGGGAA
        361 CATAGTCCCA GCTATTCACC AAATCCATCA ATTTCCTCTC ATTCATGAAG TCTATCATTC
        421 CTTCTTCTCC TCCATGTTTG CCCATGGTAA AAGAGCTGAA CATGTTGGAC AAAGCATTAG
        481 AAAGCTCATC GTAGCTTTTA TACATCCTCA AATCGATTTT CCTCAAGTAC GGTGCTCCGT
        541 CCATTGATAC CTTCACGAAC GCCGCCGCCT CCGGGCCACC GCTTGATTTT TGGCAGGAAA
        601 CCATCACGTT CTTCCGGTAT GATCTCACCG GTGGCCATCC CACAACTTGT GCCTTGGCCG
        661 GAGGTTTGGC TGGATCTTTA GGACAAGCAC TCTTCTCCTT GGAATCAAAA GTCACGACGT
        721 CATGAGTCGT AGATCCTTCC TTGTTTGCAG GCTCATTATT CAGATTTAGC TTCAGATCAA
        781 CCGTCTCTGA NAACCCTCTC TTGTTTCCGG TTACCGGANC CACTGTATCT CCACCGGGAA
        841 NANCAAGANA CAGCTCANTC TCCNTCANAN TCAGCTCNAC ACNGCCCANG GNTGNGGNCN
        901 TATNATCATC NNGNTTTTTC NAANGNAAAN CACNNNCCCN NNNNNTNCNG GGGGGNCNNN
      961 NNNNNTTTTT TNNNNCNNNN NNNNANNNNN NNNNANNNNN NNNNN
      //
      Primer: SV40pro_ori_F
      Region: SV40 pro_ori,NeoR_KanR
      Sequence file: RDB18378_B0Hhf.seq check
      >08469_18378_B0Hh_3_SV40pro_ori_F_F11_17_ABI24.ab1
          1 NNNNNNNNNN NNNNGNNGCC TCNGCCTCTG NNCTATTCNN GAAGTAGTGA GGAGGCTTTT
         61 TTGGAGGCCT AGGCTTTTGC AAAGATCGAT CAAGAGACAG GATGAGGATC GTTTCGCATG
        121 ATTGAACAAG ATGGATTGCA CGCAGGTTCT CCGGCCGCTT GGGTGGAGAG GCTATTCGGC
        181 TATGACTGGG CACAACAGAC AATCGGCTGC TCTGATGCCG CCGTGTTCCG GCTGTCAGCG
        241 CAGGGGCGCC CGGTTCTTTT TGTCAAGACC GACCTGTCCG GTGCCCTGAA TGAACTGCAA
        301 GACGAGGCAG CGCGGCTATC GTGGCTGGCC ACGACGGGCG TTCCTTGCGC AGCTGTGCTC
        361 GACGTTGTCA CTGAAGCGGG AAGGGACTGG CTGCTATTGG GCGAAGTGCC GGGGCAGGAT
        421 CTCCTGTCAT CTCACCTTGC TCCTGCCGAG AAAGTATCCA TCATGGCTGA TGCAATGCGG
        481 CGGCTGCATA CGCTTGATCC GGCTACCTGC CCATTCGACC ACCAAGCGAA ACATCGCATC
        541 GAGCGAGCAC GTACTCGGAT GGAAGCCGGT CTTGTCGATC AGGATGATCT GGACGAAGAG
        601 CATCAGGGGC TCGCGCCAGC CGAACTGTTC GCCAGGCTCA AGGCGAGCAT GCCCGACGGC
        661 GAGGATCTCG TCGTGACCCA TGGCGATGCC TGCTTGCCGA ATATCATGGT GGAAAATGGC
        721 CGCTTTTCTG GATTCATCGA CTGTGGCCGG CTGGGTGTGG CGGANCGCTA TCANGACATA
        781 GCGTTGGCTA CNNGTGATAT TGCTGAAGAN CTTGGCGGCG AATGGGCTGA CCGCTTCCTC
        841 GNGCTTTACN GNATCGCCGC TCCCNANTCN CANCGCATCG CCTTCNATCG CNNTCNNGAN
      901 NANNNNNTCN NNNCGGNNNN NNNGGNNNNC NAAANNNNNN NNNN
      //

      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.
      user report Miura, K., Application of auxin-inducible degron technology to mouse oocyte activation with PLCzeta. J. Reprod. Dev. 64 (4): 319-326 (2018). PMID 29731504.

      2021.11.02

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