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pMK290 (mAID-mClover-Hygro)

A plasmid for construction of C-terminal mAID-mClover tagging donors.

Catalog number RDB13933
Resource name pMK290 (mAID-mClover-Hygro)
Alternative name mAID-mClover-Hygro, 1025
Clone info. A plasmid for construction of C-terminal mAID-mClover tagging donors. Please refer Fig. 4 of Natsume, T., Cell Reports 15, 210-218 (2016) for an instruction about how to use this clone.
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 pBluescriptII KS- (Plasmid)
Size of vector backbone 3.0 kb
Selectable markers Am^r
Gene/insert name Arabidopsis thaliana AtIAA17 cDNA Aequorea victoria GFP cDNA
Depositor Kanemaki, Masato |
 
Other clones in our bank Arabidopsis thaliana AtIAA17 (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 designated by the DEPOSITOR is requested (Natsume, T., Cell Reports, 15: 210-218, 2016). 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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Schedule A (GFP-A) [Link]
Remarks
Please use Schedule A (GFP-A) additionally.
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RDB13933 pMK290 (mAID-mClover-Hygro) DNA solution

cDNA Request [in Japanese] [in English]

References and tips

Electronic file

Featured content

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

References

original Natsume, T., Rapid protein depletion in human cells by auxin-inducible degron tagging with short homology donors. Cell Rep. 15 (1): 210-218 (2016). PMID 27052166.

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 RDB13933_A6B8p1_v2.pdf

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

Primer: M13_-40 RDB13933_A6B8a_2.seq
>D03070A17_A6B8_1_M13(-40)_B05_05.ab1
    1 CCGCCGCTAA CGCGCGTATA CGACTCACTA TAGGGCGAAT TGGAGCTCCC CGGGATCCGG
   61 TGCAGGCGCC AAGGAGAAGA GTGCTTGTCC TAAAGATCCA GCCAAACCTC CGGCCAAGGC
  121 ACAAGTTGTG GGATGGCCAC CGGTGAGATC ATACCGGAAG AACGTGATGG TTTCCTGCCA
  181 AAAATCAAGC GGTGGCCCGG AGGCGGCGGC GTTCGTGAAG GTATCAATGG ACGGAGCACC
  241 GTACTTGAGG AAAATCGATT TGAGGATGTA TAAAGCTAGC ATGGTGAGCA AGGGCGAGGA
  301 GCTGTTCACC GGGGTGGTGC CCATCCTGGT CGAGCTGGAC GGCGACGTAA ACGGCCACAA
  361 GTTCAGCGTC CGCGGCGAGG GCGAGGGCGA TGCCACCAAC GGCAAGCTGA CCCTGAAGTT
  421 CATCTGCACC ACCGGCAAGC TGCCCGTGCC CTGGCCCACC CTCGTGACCA CCTTCGGCTA
  481 CGGCGTGGCC TGCTTCAGCC GCTACCCCGA CCACATGAAG CAGCACGACT TCTTCAAGTC
  541 CGCCATGCCC GAAGGCTACG TCCAGGAGCG CACCATCTCT TTCAAGGACG ACGGTACCTA
  601 CAAGACCCGC GCCGAGGTGA AGTTCGAGGG CGACACCCTG GTGAACCGCA TCGAGCTGAA
  661 GGGCATCGAC TTCAAGGAGG ACGGCAACAT CCTGGGGCAC AAGCTGGAGT ACAACTTCAA
  721 CAGCCACAAC GTCTATATCA CGGCCGACAA GCAGAAGAAC GGCATCAAGG CTAACTTCAA
  781 GATCCGCCAC AACGTTGAGG ACGGCAGCGT GCAGCTCGGC CGACCACTAC CAGCAGAACA
  841 CCCCCATCGG CGACGGCCCC GTGCTGCTGC CCGACAACCA CTACCTGAGC CATCAGTCCA
  901 AGCTGAGCAA AGACCCCAAC GAGAAGCGCG AATCACATGG GTCCCTGCTG GGAGTTCGTG
  961 ACCGCCCGCC CGGGATTACC ACTTGGGCAA TGGGAACGAG CTTGTACCAG GTTAATCCTA
 1021 GATAAACTGA TCATAAATCC AGCGCATAAC CAACAATTTG GTAGAAGGTT TTACTTGGCT
 1081 TAAAAAAACC CTCCCACAAC CTTCCACCCT GAACCTGAAA CACATAAAAA TGAATTGCCA
1141 ACTGTGGTTG TTAAACCTTG GATTATATTG GCCAGCCTTT AATATATGCG AGTA
//
Primer: PGK_N_rev RDB13933_A6B8b_2.seq
>D03070A17_A6B8_1_PGK-N-rev_E06_15.ab1
    1 CATGGGGAAC GCCTCCCTAC CGGTAGAATT TCGAGGTCGA GATCCTAAGC TTGGCTGGAC
   61 GTAAACTCCT CTTCAGACCT AATAACTTCG TATAGCATAC ATTATACGAA GTTATATTAA
  121 GGGTTATTGA ATATGATCGA CGCGTTAAGA TACATTGATG AGTTTGGACA AACCACAACT
  181 AGAATGCAGT GAAAAAAATG CTTTATTTGT GAAATTTGTG ATGCTATTGC TTTATTTGTA
  241 ACCATTATAA GCTGCAATAA ACAAGTTAAC AACAACAATT GCATTCATTT TATGTTTCAG
  301 GTTCAGGGGG AGGTGTGGGA GGTTTTTTAA AGCAAGTAAA ACCTCTACAA ATGTGGTATG
  361 GCTGATTATG ATCAGTTATC TAGATTACTT GTACAGCTCG TCCATGCCAT GTGTAATCCC
  421 GGCGGCGGTC ACGAACTCCA GCAGGACCAT GTGATCGCGC TTCTCGTTGG GGTCTTTGCT
  481 CAGCTTGGAC TGATGGCTCA GGTAGTGGTT GTCGGGCAGC AGCACGGGGC CGTCGCCGAT
  541 GGGGGTGTTC TGCTGGTAGT GGTCGGCGAG CTGCACGCTG CCGTCCTCAA CGTTGTGGCG
  601 GATCTTGAAG TTAGCCTTGA TGCCGTTCTT CTGCTTGTCG GCCGTGATAT AGACGTTGTG
  661 GCTGTTGAAG TTGTACTCCA GCTTGTGCCC CAGGATGTTG CCGTCCTCCT TGAAAGTCGA
721 TGCCCTTCAG CTCGATGCGG TTCACCAGGG GTGTCGCCCT
//
Primer: PGKpro_F RDB13933_A6B8c_2.seq
>D03070A17_A6B8_1_PGK-proF_H07_22.ab1
    1 ATCGGCGTAT CTCTCTTCTC ATCTCCGGGC CTTTCGACCT GCATCCATCT AGATCTCGAG
   61 CAGCTGAAGC TTACCGCTAG CATGGATAGA TCCGGAAAGC CTGAACTCAC CGCGACGTCT
  121 GTCGAGAAGT TTCTGATCGA AAAGTTCGAC AGCGTCTCCG ACCTGATGCA GCTCTCGGAG
  181 GGCGAAGAAT CTCGTGCTTT CAGCTTCGAT GTAGGAGGGC GTGGATATGT CCTGCGGGTA
  241 AATAGCTGCG CCGATGGTTT CTACAAAGAT CGTTATGTTT ATCGGCACTT TGCATCGGCC
  301 GCGCTCCCGA TTCCGGAAGT GCTTGACATT GGGGAATTCA GCGAGAGCCT GACCTATTGC
  361 ATCTCCCGCC GTGCACAGGG TGTCACGTTG CAAGACCTGC CTGAAACCGA ACTGCCCGCT
  421 GTTCTGCAGC CGGTCGCGGA GGCCATGGAT GCGATCGCTG CGGCCGATCT TAGCCAGACG
  481 AGCGGGTTCG GCCCATTCGG ACCGCAAGGA ATCGGTCAAT ACACTACATG GCGTGATTTC
  541 ATATGCGCGA TTGCTGATCC CCATGTGTAT CACTGGCAAA CTGTGATGGA CGACACCGTC
  601 AGTGCGTCCG TCGCGCAGGC TCTCGATGAG CTGATGCTTT GGGCCGAGGA CTGCCCCGAA
  661 GTCCGGCACC TCGTGCACGC GGATTTCGGC TCCAACAATG TCCTGACGGA CAATGGCCGC
  721 ATAACAGCGG TCATTGACTG GAGCGAGGCG ATGTTCGGGG ATTCCCAATA CGAGGTCGCC
  781 AACATCTTCT TCTGGAGGCC GTGGTTGGCT TGTATGGAGC AGCAGACGCG CTACTTCGAG
  841 CGGAGGCATC CGGAGCTTGC AGATCGCCGC GGCTCCGGGC GTATATGCTC CGCATTGGTC
  901 TTGACCAACT CTATCAGAGC TTGGTTTGAC GGCAATTTCG ATGATGCAGC TTGGGCGCAG
  961 GGTCGATGCG ACGCAATCGT TCCGATCCGG AGCCCGGGAC TGTCCGGGCG TACACAAATT
 1021 CGCCGCCAGA AGCGCGGCCC GTCTGGACCG ATGCTGGTGT AGAAGTTACT CCGCCGGAAT
 1081 AGTGGAATCG GACGTCCCCA GCCACTCGTT CGAAGGTCTA GATAGCCTAG CCGCCCGCTC
1141 CACGAACCCG CTAGCGCGTC
//
Primer: M13R RDB13933_A6B8d_2.seq
>D03070A17_A6B8_1_M13R_B04_04.ab1
    1 CAGGCCGGCA TTACCCTCAC TAAAGGGAAC AAAAGCTGGG TACCGGATCC TAGTGAACCT
   61 CTTCGAGGGA CCTAATAACT TCGTATAGCA TACATTATAC GAAGTTATAT TAAGGGTTCC
  121 GGATCTCGAC CAGCTTCTGA TGGAATTAGA ACTTGGCAAA ACAATACTGA GAATGAAGTG
  181 TATGTGGAAC AGAGGCTGCT GATCTCGTTC TTCAGGCTAT GAAACTGACA CATTTGGAAA
  241 CCACAGTACT TAGAACCACA AAGTGGGAAT CAAGAGAAAA ACAATGATCC CACGAGAGAT
  301 CTATAGATCT ATAGATCATG AGTGGGAGGA ATGAGCTGGC CCTTAATTTG GTTTTGCTTG
  361 TTTAAATTAT GATATCCAAC TATGAAACAT TATCATAAAG CAATAGTAAA GAGCCTTCAG
  421 TAAAGAGCAG GCATTTATCT AATCCCACCC CACCCCCACC CCCGTAGCTC CAATCCTTCC
  481 ATTCAAAATG TAGGTACTCT GTTCTCACCC TTCTTAACAA AGTATGACAG GAAAAACTTC
  541 CATTTTAGTG GACATCTTTA TTGTTTAATA GATCATCAAT TTCTGCATCC CGGGGATCTG
  601 ATATCATCGA TGCATGGGGT CGTGCGCTCC TTTCGGTCGG GCGCTGCGGG TCGTGGGGCG
  661 GGCGGCTAGC CTATTCCTTT GCCCTCGGAC GAGTGCTGGG GCGTCGGTTT CCACTATCGG
  721 CGAGTACTTC TACACAGCCA TCGGTCCAGA CGGCCGCGCT TCTGCGGGCG ATTTGTGTAC
  781 GCCCGACAGT CCCGGCTCCG GATCGGACGA TTGCGTCGCA TCGACCCTGC GCCCAAGCTG
  841 CATCATCGAA ATTGCCGTCA ACCAAGCTCT GATAGAGTTG GTCAAGACCA ATGCGGAGCA
  901 TATACGCCCG GAGCCGCGGC GATCCTGCAA GCTCCGGATG CCTCCGCTCG AAGTAGCGCG
  961 TCTGCTGCTC CATACAAGCC AACCCACGGC CTCCAGAAGA AGATGTTGGG CGGACCTCGT
 1021 ATTGGGGAAA TCCCCCGAAC ATCGCCTCGC CTCCAGTCAA ATGACCCGCT GGTATGGCCG
 1081 GCCATGGTCG TCAGGGACAT TGGTGGAGTG AATCCGCGGT GCACGAAGGG TGGCCGGAAC
1141 CTTCCGGGGG GC
//

Please visit Sequencing and PCR primers for primer information.


References

References

original Natsume, T., Rapid protein depletion in human cells by auxin-inducible degron tagging with short homology donors. Cell Rep. 15 (1): 210-218 (2016). PMID 27052166.
reference Goto, H., Chk1-mediated Cdc25A degradation as a critical mechanism for normal cell cycle progression. J Cell Sci. 2019 Jan 25;132(2). pii: jcs223123 (2019). PMID 30635443.
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 Takagi, M., Perichromosomal protein Ki67 supports mitotic chromosome architecture. Genes Cells 21 (10): 1113-1124 (2016). PMID 27610954.
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 .

2019.03.08

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