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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 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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RDB08469 pMK106 DNA solution

cDNA Request [in Japanese] [in English]

References and tips

Electronic file

Electronic file Sequence RDB08469z.seq from Depositor
Electronic file PDF RDB08469.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 RDB08469_A5Cpp1-2.pdf

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

Primer: CMV-Forward RDB08469_A5Cpa.seq
>08469_08469_A5Cp_CMV-Forward_E11_14.ab1
    1 TTTTACAAAT CCGCCCATTG ACGCAAATGG GCGGTAGGCG TGTACGGTGG GAGGTCTATA
   61 TAAGCAGAGC TGGTTTAGTG AACCGTCAGA TCCGCTAGCG CTACCGGACT CAGATCTCGA
  121 GATGACGTAC TTCCCGGAGG AGGTGGTGGA GCACATCTTC AGCTTCCTGC CGGCGCAGCG
  181 CGACCGCAAC ACGGTCTCGC TCGTCTGCAA GGTGTGGTAC GAGATCGAGA GGCTGAGCCG
  241 CCGCGGCGTC TTCGTGGGCA ACTGCTACGC CGTGCGCGCC GGCCGCGTCG CCGCGCGGTT
  301 CCCCAACGTG CGGGCGCTCA CGGTGAAGGG GAAGCCCCAC TTCGCCGACT TCAACCTCGT
  361 GCCCCCCGAC TGGGGCGGCT ACGCGGGGCC GTGGATCGAG GCGGCCGCGA GGGGATGCCA
  421 CGGCCTGGAG GAGCTCAGGA TGAAGCGGAT GGTGGTGTCC GACGAGAGCC TCGAGCTGCT
  481 GGCTCGCTCG TTCCCGCGGT TCAGGGCTCT TGTTCTTATC AGCTGCGAGG GGTTCAGCAC
  541 TGACGGGCTA GCCGCCGTCG CGAGCCATTG CAAGCTTCTG AGGGAGTTGG ATTTGCAGGA
  601 AAATGAAGTG GAGGATCGAG GGCCTAGGTG GCTTTCCTGC TTCCCTGATT CCTGCACATC
  661 ACTTGTCTCA TTGAATTTTG CCTGCATCAA AGGGGAGGTT AATGCTGGTT CACTGGAGAG
  721 ACTTGTTAGC AGGTCCCCAA ACCTGCGGAG TTTGAGGCTG AATCGATCTG TATCGGTAGA
  781 TACACTTGCA AAGATACTAC TGCGTACCCC TAACTTGGAG GATTTGGGGA CAGGGAATTT
  841 GACAGATGAC TTCCAAACTG AGTCCTACTT TAAGCTTACC AGTGCTCTGG AGAAATGCAA
  901 GATGTTGAGG AGTTTGTCTG GATTCTGGGA TGCTTCTCCT GTTTGCCTGT CATTTATCTA
  961 CCCCCTGTGT GCTCAACTGA CAGGATTGAA CTTGAGCTAT GCACCCACAC TTGATGCTTC
 1021 TGACCTTTAC AAAAATGATT AGCCGCTGTT GTGAAGCTCC CAACGCCCTT TGGGTTACTT
 1081 GATGTATCTC GGACAGGGCT GGCCAGTGTG ACCTCCAAGT TGCAAAGACT TTGCCCAGGA
1141 AACCCTCCCA GGG
//
Primer: ME-1290Rv RDB08469_A5Cpb.seq
>08469_08469_A5Cp_ME-1290Rv_F11_17.ab1
    1 ATGTCAAGTC ATTTTATGTT TCAGGTTCAG GGGGAGGTGT GGGAGGTTTT TTAAAGCAAG
   61 TAAAACCTCT ACAAATGTGG TATGGCTGAT TATGATCTAG AGTCGCGGCC GCAGTGATCA
  121 CAGTACTCTA CAAGAGATTC TGAATGGAGA TATCTTTCTC GAGCTGTAGA ATAATTGCTC
  181 TTAAAGATGG ATCTTTCGCC CAGTTGATTT GGCATCCGAC TATGATATTC TGGAGTATAT
  241 TTTGAACTAA CGTAGTCACC TTCCTTTCAT ACTGTACATT CCTATGGATA AACTTCAACT
  301 TTTCAGCAAT TTTCATTCTG ACTACTTCCT TTTTTTTCCT CACTGTGGTA AAAATTCGCA
  361 GCGCCTTCTC TCCTTCGGCC TGTTTCAAAA AGTATCTTTT CCTTCTAATG TCATTCAAAT
  421 TCTCTCTAAT CACACGTGCT TGCTGATGAG CTTGAATTAT TTCCCAGTTG AGCTTGATTA
  481 TGTCTCTCAT TGTTTCCACG ATCAGTCGTG CGCAACCATC ATTTTCTTTC ACTCTCTTTC
  541 TCAGAGCATC CATAAGCTGG ATCCTTTTTA TGACTAATGC TTTGCTTTCA AAGGCACTTT
  601 TTACTTTCCC CACTTCTTCC AAAGTTTCTT TGGCACTTTC GATAAAGTTT ACTTCCGCGC
  661 CACTGTTGCA CCCTTCATCG GGGGGGCACG CGGTGTCGAA CTCCATCACC AGCTGCTTCA
  721 GATGCGTCCT CGCGCAGAGG TGCTCCAGGA CGTCCCGCTT CGCGTCCGGA TCGGCCGCCG
  781 TCTCGGCCTC AGCGCCCGGC CCGGACCCGA CGGACTCCGA CAGCCGCCCC GCCATGGTGA
  841 ATTCGATATC GGCACCCGCT CCAGCGCCTG CACCAGCTCC GGGCCGCAGC TCTGCTCTTG
  901 CACTTCTCCA TCGCCCTCGG AGCGAGACCA ATGGCATCCG ATCCTTTCAT GAGACGTAAA
  961 CGCTTGCATG TATCGACGAA CATTGGCCAA GGAACGTCGC CGACGAGCAT CCAATCACCG
 1021 TCTTTGTCTT CAATAAGAGG GAACAATAGT TCCCAGCCTA TTTCACCAAA ATCATCATTT
 1081 CTTCTCATTC AATGAAGTTC TATTCATTCC TTCATCTCCT CCATGTTTGC CCATGGTTAA
1141 AGAGCTGACA TGGTTGGAAC AAAGCATTAG AAAGCCTTCA ATCCGTAGCC TTTTTTAT
//
Primer: IRES-R RDB08469_A5Cpc.seq
>08469_08469_A5Cp_IRES-R_H11_23.ab1
    1 GCATAGCGGC TTCGGCAGTA CGTTAGGGGG GGGGGAGGGA GAGGGGCGGA TCCCGGGGAT
   61 CTCTTGAATG ATCGTTCCAC TTTTTAGCTA GTGGATCCGT TCAAGTCTTC TTCTGAGATT
  121 AATTTTTGTT CACCGTTCAA GTCTTCCTCG GAGATTAGCT TTTGTTCACC GTTCAAATCT
  181 TCTTCAGAAA TCAACTTTTG TTCACCTCTA GAGGATCCGT TCAAGTCTTC TTCTGAGATT
  241 AATTTTTGTT CACCGTTCAA GTCTTCCTCG GAGATTAGCT TTTGTTCACC GTTCAAATCT
  301 TCTTCAGAAA TCAACTTTTG TTCACCTCTA GAGGATCCGT TCAAGTCTTC TTCTGAGATT
  361 AATTTTTGTT CACCGTTCAA GTCTTCCTCG GAGATTAGCT TTTGTTCACC GTTCAAATCT
  421 TCTTCAGAAA TCAACTTTTG TTCACCACTA GCAGCAGAAC CGGAGTCGAC TAGGATTTTA
  481 ACAAAATTTG GTGCATCATC CCTCGCCCCA GCAGTTGTGC GGTACACATA TAATTTCTCC
  541 ACTTTAGGCA GATCTCCATG GTTTTCCTCC ATTTCATTGC TACCATCCCG CTCATTTATG
  601 ACCTCAACAT TGAGCATCGG CATCTTTGAC GCAAGGACTT GGCACCCCTT TAACGTGACA
  661 TTGCAAGATG ACATCCAAAG GGATCGCATT GTCTCGTACC TAGCAAAATT CCCCAAGAGT
  721 GCAGCATCAC CAAACGGGCT ATCTCTTATC TCCAGTTTCC TTAAATTCTT GCATCCATTC
  781 ATAACATGCA TCATACCCTT ATCACTGTCA CCAGCAAATG CTATAGAAAG CATCTCAAGT
  841 TGTTTTGCAT ATTTCCCAAT ATACATGAAA ACTTTGTCTG TGAGAAGACC AGATATTGAC
  901 AAACGTTGTA ATCCCTTGCA CTCACGAACA ATAGCTCCAA AGCCTTCATC TAATGGTTTG
  961 GCTTGTCACA ACATCAGGCT TCCCTGGCTC AAGAATACAA AGTCTGAATC GTGTGAAATT
 1021 TGGGACAGTT CTTGGCGACA GTAACTAGTG CAGCATTTAG TCATTTGGTG ACAGAAGTAC
 1081 AGTAGTGAGT TCAGTTTTGG ACAGCCAAGG ATACTGCACA GTCCTCCTCT GGTCACTGCC
1141 AGAATACAGC CTACGTAGAA TCTTGGAATG GAAAAAAATA TCA
//
Primer: IRES-R RDB08469_A5Cpd.seq
>08469_08469_A5Cp_IRES-F_G11_20.ab1
    1 GCATCTTTCT GTGTTTAGTC GAGGTTAAAA AAACGTCTAG GCCCCCCGAA CCACGGGGAC
   61 GTGGTTTTCC TTTGAAAAAC ACGATGATAA TATGGCCACA ACCATGGGCA GTGTCGAGCT
  121 GAATCTGAGG GAGACTGAGC TGTGTCTTGG TCTTCCCGGT GGAGATACAG TGGCTCCGGT
  181 AACCGGAAAC AAGAGAGGGT TCTCAGAGAC GGTTGATCTG AAGCTAAATC TGAATAATGA
  241 GCCTGCAAAC AAGGAAGGAT CTACGACTCA TGACGTCGTG ACTTTTGATT CCAAGGAGAA
  301 GAGTGCTTGT CCTAAAGATC CAGCCAAACC TCCGGCCAAG GCACAAGTTG TGGGATGGCC
  361 ACCGGTGAGA TCATACCGGA AGAACGTGAT GGTTTCCTGC CAAAAATCAA GCGGTGGCCC
  421 GGAGGCGGCG GCGTTCGTGA AGGTATCAAT GGACGGAGCA CCGTACTTGA GGAAAATCGA
  481 TTTGAGGATG TATAAAAGCT ACGATGAGCT TTCTAATGCT TTGTCCAACA TGTTCAGCTC
  541 TTTTACCATG GGCAAACATG GAGGAGAAGA AGGAATGATA GACTTCATGA ATGAGAGGAA
  601 ATTGATGGAT TTGGTGAATA GCTGGGACTA TGTTCCCTCT TATGAAGACA AAGACGGTGA
  661 TTGGATGCTC GTCGGCGACG TTCCTTGGCC AATGTTCGTC GATACATGCA AGCGTTTACG
  721 TCTCATGAAA GGATCGGATG CCATTGGTCT CGCTCCGAGG GCGATGGAGA AGTGCAAGAG
  781 CAGAGCTGCG GCCGGAGCTG GTGCAGGCGC TGGAGCGGGT GCCGATATCG AATTCACCAT
  841 GGCGGGGCGG CTGTCGGAGT CCGTCGGGTC CGGGCCGGGC GCTGAGGCCG AGACGGCGGC
  901 CGATCCGGAC GCGAAGCGGG ACGTCCTGTA GCACCTCTGC GCGAGGACGC ATCTGAAGCA
  961 GCTGGTGATG GAGTTCGACA CCGCGTGCCC CCCCGATGAA GGGTGCAACA GTGGGCGCGG
 1021 AAGTAAACTT TATCGAAAGT GCCCAAAGAA ACTTTGGAAG AAGTGGGGGA AAAGTAAAAA
 1081 GTGCTTTTGA AGCAAGCATA GTCAATAAAG GATCCAGCTA ATGATGCTCT GAGAAGAGAG
1141 TGAAAGCAAA TGATGGGTTG CGC
//
Primer: SV40proF_V2 RDB08469_A5Cpe.seq
>08469_08469_A5Cp_SV40proF_V2_A12_03.ab1
    1 GTCGAGCTTT TTTGGAGCTA GGCTTTTGCA AGATCGATCA AGAGACAGGA TGAGGATCGT
   61 TTCGCATGAT TGAACAAGAT GGATTGCACG CAGGTTCTCC GGCCGCTTGG GTGGAGAGGC
  121 TATTCGGCTA TGACTGGGCA CAACAGACAA TCGGCTGCTC TGATGCCGCC GTGTTCCGGC
  181 TGTCAGCGCA GGGGCGCCCG GTTCTTTTTG TCAAGACCGA CCTGTCCGGT GCCCTGAATG
  241 AACTGCAAGA CGAGGCAGCG CGGCTATCGT GGCTGGCCAC GACGGGCGTT CCTTGCGCAG
  301 CTGTGCTCGA CGTTGTCACT GAAGCGGGAA GGGACTGGCT GCTATTGGGC GAAGTGCCGG
  361 GGCAGGATCT CCTGTCATCT CACCTTGCTC CTGCCGAGAA AGTATCCATC ATGGCTGATG
  421 CAATGCGGCG GCTGCATACG CTTGATCCGG CTACCTGCCC ATTCGACCAC CAAGCGAAAC
  481 ATCGCATCGA GCGAGCACGT ACTCGGATGG AAGCCGGTCT TGTCGATCAG GATGATCTGG
  541 ACGAAGAGCA TCAGGGGCTC GCGCCAGCCG AACTGTTCGC CAGGCTCAAG GCGAGCATGC
  601 CCGACGGCGA GGATCTCGTC GTGACCCATG GCGATGCCTG CTTGCCGAAT ATCATGGTGG
  661 AAAATGGCCG CTTTTCTGGA TTCATCGACT GTGGCCGGCT GGGTGTGGCG GACCGCTATC
  721 AGGACATAGC GTTGGCTACC CGTGATATTG CTGAAGAGCT TGGCGGCGAA TGGGCTGACC
  781 GCTTCCTCGT GCTTTACGGT ATCGCCGCTC CCGATTCGCA GCGCATCGCC TTCTATCGCC
  841 TTCTTGACGA GTTCTTCTGA GCGGGACTCT GGGGTTCGAA ATGACCGACC AAGCGACGCC
  901 CAACCTGCCA TCACGAGATT TCGATTCCAC CGCCGCCTTC TATGAAAGGT TGGGCTTCGG
  961 AATCGTTTTC CCGGGACGCC GGCTGGATGA TCCTCCAGCG CGGGGATCTC ATGCTGGGAG
 1021 TTCTTCGCCC ACCCTAGGGG GAGGCTAACT GAAACACGGA GGAGACATAC CCGGAAGGGA
 1081 ACCCGCGCCT ATGACGGGCA TAAAAAGACG ATTAACGCAC GTGATGGGTT CGGTTGATCA
1141 TTAACGCGGG GATCGGTTCC CAAGGGGCCT GTGG
//

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.
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.

2019.04.02

GNP_filter3_RDBDEP_html_190327.pl