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pMK289 (mAID-mClover-NeoR)

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

Catalog number RDB13932
Resource name pMK289 (mAID-mClover-NeoR)
Alternative name mAID-mClover-NeoR, 1002
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. Please note that this plasmid does not contain the loxP sites, which are intended for removal of the marker.
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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RDB13932 pMK289 (mAID-mClover-NeoR) 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 RDB13932_A6B8p1_v2.pdf

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

Primer: M13_-40 RDB13932_A6B8a_2.seq
>D03070A16_A6B8_1_M13(-40)_A05_02.ab1
    1 AAGGCAGCTA 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 GCAGCTCGCC GACCACTACC AGCAGAACAC
  841 CCCCATCGGC GACGGCCCCG TGCTGCTGCC CGACAACCAC TACCTGAGCC ATCAGTCCAA
  901 GCTGAGCAAA GACCCCAACG AGAAGCGCGA TCACATGGTC CTGCTGGAGT TCGTGACCGC
  961 CGCCCGGGAT TACACATGGC ATGGAACGAG CTGTACAAGT AATCTAGATA ACTGATCATA
 1021 TCAGCCCATA CCACATTTGT AGAGGTTTTA CTGCTTAAAA CCTCCCACAC TCCCCTTGAA
 1081 CTTGAAACAT AAAATGAATG CCATGATGGT GCTACCTGTC TATGCGACTT AAAATGGTCA
1141 TAGCCATAGC CATTCACGAT
//
Primer: SV40polyA-R RDB13932_A6B8b_2.seq
>D03070A16_A6B8_1_SV40polyA-R_H04_22.ab1
    1 TCCCCCCCCC CGGTATTTAG CAGTAACCTC TACAATGTTG GTATGGGCTG ATTATGATCA
   61 GTTATCTAGA TTACTTGTAC AGCTCGTCCA TGCCATGTGT AATCCCGGCG GCGGTCACGA
  121 ACTCCAGCAG GACCATGTGA TCGCGCTTCT CGTTGGGGTC TTTGCTCAGC TTGGACTGAT
  181 GGCTCAGGTA GTGGTTGTCG GGCAGCAGCA CGGGGCCGTC GCCGATGGGG GTGTTCTGCT
  241 GGTAGTGGTC GGCGAGCTGC ACGCTGCCGT CCTCAACGTT GTGGCGGATC TTGAAGTTAG
  301 CCTTGATGCC GTTCTTCTGC TTGTCGGCCG TGATATAGAC GTTGTGGCTG TTGAAGTTGT
  361 ACTCCAGCTT GTGCCCCAGG ATGTTGCCGT CCTCCTTGAA GTCGATGCCC TTCAGCTCGA
  421 TGCGGTTCAC CAGGGTGTCG CCCTCGAACT TCACCTCGGC GCGGGTCTTG TAGGTACCGT
  481 CGTCCTTGAA AGAGATGGTG CGCTCCTGGA CGTAGCCTTC GGGCATGGCG GACTTGAAGA
  541 AGTCGTGCTG CTTCATGTGG TCGGGGTACC GGCTGAAGCA GGCCACGCCG TAGCCGAAGG
  601 TGGTCACGAG GTGGGCCAGG GCACGGGCAG CTTGCCGGTG GTGCAGATGA ACTTCAGGGT
661 CAGCTTGCCG TTG
//
Primer: SV40pro_F_V2 RDB13932_A6B8c_2.seq
>D03070A16_A6B8_1_SV40pro_F_V2_E10_13.ab1
    1 GGGGCAGCTT TTGGAGGCCT AGGCTTTTGC AAGATCGATC AAGAGACAGG ATGAGGATCG
   61 TTTCGCATGA TTGAACAAGA TGGATTGCAC GCAGGTTCTC CGGCCGCTTG GGTGGAGAGG
  121 CTATTCGGCT ATGACTGGGC ACAACAGACA ATCGGCTGCT CTGATGCCGC CGTGTTCCGG
  181 CTGTCAGCGC AGGGGCGCCC GGTTCTTTTT GTCAAGACCG ACCTGTCCGG TGCCCTGAAT
  241 GAACTGCAAG ACGAGGCAGC GCGGCTATCG TGGCTGGCCA CGACGGGCGT TCCTTGCGCA
  301 GCTGTGCTCG ACGTTGTCAC TGAAGCGGGA AGGGACTGGC TGCTATTGGG CGAAGTGCCG
  361 GGGCAGGATC TCCTGTCATC TCACCTTGCT CCTGCCGAGA AAGTATCCAT CATGGCTGAT
  421 GCAATGCGGC GGCTGCATAC GCTTGATCCG GCTACCTGCC CATTCGACCA CCAAGCGAAA
  481 CATCGCATCG AGCGAGCACG TACTCGGATG GAAGCCGGTC TTGTCGATCA GGATGATCTG
  541 GACGAAGAGC ATCAGGGGCT CGCGCCAGCC GAACTGTTCG CCAGGCTCAA GGCGAGCATG
  601 CCCGACGGCG AGGATCTCGT CGTGACCCAT GGCGATGCCT GCTTGCCGAA TATCATGGTG
  661 GAAAATGGCC GCTTTTCTGG ATTCATCGAC TGTGGCCGGC TGGGTGTGGC GGACCGCTAT
  721 CAGGACATAG CGTTGGCTAC CCGTGATATT GCTGAAGAGC TTGGCGGCGA ATGGGCTGAC
  781 CGCTTCCTCG TGCTTTACGG TATCGCCGCT CCCGATTCGC AGCGCATCGC CTTCTATCGC
  841 CTTCTTGACG AGTTCTTCTG AGCGGGACTC TGGGGTTCGA AATGACCGAC CAAGCGACGC
  901 CCAACCTGCC ATCACGAGAT TTCGATTCCA CCGCCGCCTT CTATGAAAGG TTGGGCTTCG
  961 GAATCGTTTT CCGGGACGCC CGGCTGGATG ATCCCTCCAG CGCGGGGATC TCATGCTGGG
 1021 AGTTCTTTCG CCCACCCTAG GGGAGGCTAA CTGAAACACG GAAGAGACAA TACCGGAAAG
1081 ACCGCGCTAT GACGGCCAAT AAAAGGACAG GAAATAAAAC CGCCCACCG
//
Primer: M13R RDB13932_A6B8d_2.seq
>D03070A16_A6B8_1_M13R_A04_01.ab1
    1 CTGGGCGCAA TTAACCTCAC TAAAGGGAAC AAAAGCTGGG TACCGGATCC AACGACCCAA
   61 CACCGTGCGT TTTATTCTGT CTTTTTATTG CCGTCATAGC GCGGGTTCCT TCCGGTATTG
  121 TCTCCTTCCG TGTTTCAGTT AGCCTCCCCC TAGGGTGGGC GAAGAACTCC AGCATGAGAT
  181 CCCCGCGCTG GAGGATCATC CAGCCGGCGT CCCGGAAAAC GATTCCGAAG CCCAACCTTT
  241 CATAGAAGGC GGCGGTGGAA TCGAAATCTC GTGATGGCAG GTTGGGCGTC GCTTGGTCGG
  301 TCATTTCGAA CCCCAGAGTC CCGCTCAGAA GAACTCGTCA AGAAGGCGAT AGAAGGCGAT
  361 GCGCTGCGAA TCGGGAGCGG CGATACCGTA AAGCACGAGG AAGCGGTCAG CCCATTCGCC
  421 GCCAAGCTCT TCAGCAATAT CACGGGTAGC CAACGCTATG TCCTGATAGC GGTCCGCCAC
  481 ACCCAGCCGG CCACAGTCGA TGAATCCAGA AAAGCGGCCA TTTTCCACCA TGATATTCGG
  541 CAAGCAGGCA TCGCCATGGG TCACGACGAG ATCCTCGCCG TCGGGCATGC TCGCCTTGAG
  601 CCTGGCGAAC AGTTCGGCTG GCGCGAGCCC CTGATGCTCT TCGTCCAGAT CATCCTGATC
  661 GACAAGACCG GCTTCCATCC GAGTACGTGC TCGCTCGATG CGATGTTTCG CTTGGTGGTC
  721 GAATGGGCAG GTAGCCGGAT CAAGCGTATG CAGCCGCCGC ATTGCATCAG CCATGATGGA
  781 TACTTTCTCG GCAGGAGCAA GGTGAGATGA CAGGAGATCC TGCCCCGGCA CTTCGCCCAA
  841 TAGCAGCCAG TCCCTTCCCG CTTCAGTGAC AACGTCGAGC ACAGCTGCGC AAGGAACGCC
  901 CCGTCGTGGC CCAGCCACGA TAGCCGCGCT GCCTCGTCTT GCAGTTCATT CAAGGCACCG
  961 GGACAGGGTC GGGTCTTTGA CAAAAAGAAT CCGGGGCGGC CCCTGCGCTT GACAGGCCGG
 1021 GAACCACGGG CGGGCAATCA GAAGCAGCCC GAATTGTCTG TTGGTGGCCC AGTCAATAGC
 1081 CGGAAATAGG CTCTCCACCC CAAGCCGGGC CGGGAGGAAC TGCGTGCCAA TTCCAATCCT
1141 TTGGCTCATT CAATGCCGGA AAACGGAATT CCCTTCACAT CACCTCTG
//

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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