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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 Kanemaki, Masato |
 
Other clones in our bank Gallus gallus CENPH | Oryza sativa Os04g0395600 | Arabidopsis thaliana AXR3 (KEGG orthology K14484) |

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

cDNA Request [in Japanese] [in English]

References and tips

Electronic file

Electronic file Sequence RDB08470z.seq from Depositor
Electronic file PDF RDB08470.pdf from Depositor

Featured content

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

References

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.

User reports and related articles (go to bottom)


Sequence information

RIKEN BRC has sequenced portions of this material for quality test. Primers and the results are shown below.

Data are summerized on test sheet below.

Test sheet RDB08470_A5C2p1.pdf

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

Primer: CMV-Forward RDB08470_A5C2a.seq
>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 RDB08470_A5C2b.seq
>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 RDB08470_A5C2c.seq
>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 RDB08470_A5C2d.seq
>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 RDB08470_A5C2e.seq
>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

References

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.

2018.12.11

GNP_filter3_RDBDEP_html_181012.pl