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pMK291 (mAID-mClover-Bsr) (#RDB13934)

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

Alternative name mAID-mClover-Bsr, 1024
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.
Comment 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|Developer Kanemaki, Masato |
 

Distribution information

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Terms and conditions for distribution 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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手続きの概要は、「提供申込みについて[link]」をご覧ください。
MTAに書く使用条件 利用者は、研究成果の公表にあたって寄託者の指定する文献を引用する (Natsume, T., Cell Reports, 15: 210-218, 2016)。学術研究にのみ限定、商業営利目的利用を禁ずる。学術内部利用以外の利用の際は、国立遺伝学研究所 (chizai@nig.ac.jp) に相談すること。

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RDB13934 pMK291 (mAID-mClover-Bsr) DNA solution

Please review the QC test results indicated by check icon below as well as clone information before placing your order.

How to cite this biological resource

Materials & Methods section:

The pMK291 (mAID-mClover-Bsr) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13934).

Reference section:

Natsume, T., Kiyomitsu, T., Saga, Y., Kanemaki, M.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. [link to RRC of NBRP]

Further references such as user reports and related articles (go to bottom)


QC test results

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 RDB13934_A6B8p1-3.pdf check

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

Primer: M13_-40 (Pr0410)
Region: insert mid(Linker,mAID,mClover 5')
Sequence file: RDB13934_A6B8e.seq check
>D03070A18_A6B8_1_M13(-40)_C05_08.ab1
    1 NNNNNNNNNN NNCNCNNANN NNNNTNNCNA TAGGGCGAAT TGGAGCTCCC CGGGNTCCGG
   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 GCAGAAGANC GGCATCAAGG CTAACTTCAA
  781 GATCCGCCAC AACGTTGAGG ACGGCAGCGT GCAGCTCGCC NNACCACTAC CANCNNAACA
  841 CCCCCATCGG CGANNGNCCC GNNCTGCTGC CCGANANCCA CTACCTGAGC CATCAGTNCA
  901 AGCTGANCNA AGANCCCNNC NANAAGCNNG ATCACATGGT CCNGCTNGNA NNNNNNGNNC
961 NCNGCCNGGN NTTACNNNNG GNANNNNNNA NCTGNANNAN TNNNNNNNNN N
//
Primer: M13R (Pr0160)
Region: insert 3'(loxP,mPGK ter,bsr 3')
Sequence file: RDB13934_A6B8f.seq check
>D03070A18_A6B8_1_M13R_C04_07.ab1
    1 NNNNNNNNNN NNNNNNNNNN NAANGGGNNN NNAAGCTGGG TANCGGATCC 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 TTAATTTCGG GTATATTTGA GTGGAATGAG TTCTTCAATC GTAGTTTTGA
  721 CTAACTTGCC ATTCATTTCT ATTAACACAA AACAATCTGG TGCATAGTCT GAAATCAACT
  781 CCCTACACAT ACCACAAGGA CTTACCACTC GAATACTTCT ATCTACTTCG TCAGANTAAG
  841 GGTGTCTAAC AGCTACNATC GTGTCNAAAT CCTTTTGTCC ATTNNNANCT GCNCTACCNA
  901 TCGCAANNNG CNTNNNGCAC AAACAGTTAN NCGNNCCTAT ANNCNCNTNC NATATGTACT
  961 GNCNNAAAAT GANTTNCTCN NNNTTTCNNN CNNAATTNNC CNNNTCCCNN NNNTGNATGN
1021 NNNNNNATNN NNNNAANNNN NNNNNN
//
Primer: PGK_N_rev (Pr0243)
Region: insert mid(loxP,SV40 pA)
Sequence file: RDB13934_A6B8g.seq check
>D03070A18_A6B8_1_PGK-N-rev_F06_18.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNGGNANNNN NNCGAGGTCG AGNNNCCTAA GCTTGGCTGG
   61 ANGTAAACTC CTCTTCAGAC CTAATAACTT CGTATAGCAT ACATTATACG AAGTTATATT
  121 AAGGGTTATT GAATATGATC GACGCGTTAA GATACATTGA TGAGTTTGGA CAAACCACAA
  181 CTAGAATGCA GTGAAAAAAA TGCTTTATTT GTGAAATTTG TGATGCTNTT GCNNNNNNTG
  241 TAACCATTAT AAGCTGCAAT AAACAAGTTA ACAACAACAA TTGCATTCAT TTTATGTTTC
  301 AGGTTCAGGG GGAGGTGTGG GAGGTTTTTT AAAGCNNGTA AAACCTCTAC NAATGTGGTA
  361 TGGCTGATTA TGATNNNNTA TNNNNANTAC TTGTACAGCN NGNNNNTGCC ATGTGTNNNN
  421 NCGGCGGCNG NNNNGAANNN NNNNAGGACC ATGTNATNNC GCTTCNCNNN NGNNNCTTTG
  481 CTCNNCTTGN NNNNNNGGNN NNNGTAGTGG NTGTCNGGNN NNANCACNGG GCCNNNNNNN
541 NNN
//
Primer: PGKpro_F (Pr0249)
Region: insert mid(bsr,mPGK ter 5')
Sequence file: RDB13934_A6B8h.seq check
>D03070A18_A6B8_1_PGK-proF_A08_02.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNTCCNGGC CTTTCGACCT GCATCCNTCT AGATCTCGAG
   61 CAGCTGAAGC TTACCGCTAG CATGAAAACA TTTAACATTT CTCAACAAGA TCTAGAATTA
  121 GTAGAAGTAG CGACAGAGAA GATTACAATG CTTTATGAGG ATAATAAACA TCATGTGGGA
  181 GCGGCAATTC GTACGAAAAC AGGAGAAATC ATTTCGGCAG TACATATTGA AGCGTATATA
  241 GGACGAGTAA CTGTTTGTGC AGAAGCCATT GCGATTGGTA GTGCAGTTTC GAATGGACAA
  301 AAGGATTTTG ACACGATTGT AGCTGTTAGA CACCCTTATT CTGACGAAGT AGATAGAAGT
  361 ATTCGAGTGG TAAGTCCTTG TGGTATGTGT AGGGAGTTGA TTTCAGACTA TGCACCAGAT
  421 TGTTTTGTGT TAATAGAAAT GAATGGCAAG TTAGTCAAAA CTACGATTGA AGAACTCATT
  481 CCACTCAAAT ATACCCGAAA TTAAGCTAGC CGCCCGCCCC ACGACCCGCA GCGCCCGACC
  541 GAAAGGAGCG CACGACCCCA TGCATCGATG ATATCAGATC CCCGGGATGC AGAAATTGAT
  601 GATCTATTAA ACAATAAAGA TGTCCACTAA AATGGAAGTT TTTCCTGTCA TACTTTGTTA
  661 AGAAGGGTGA GAACAGAGTA CCTACATTTT GAATGGAAGG NNTGGAGCTA CGGGGGTGGG
  721 GGTGGGGTGG GATTAGATAA ATGCCTGCTC TTTACTGAAG GCTCTTTACT ATTGCTTTAT
  781 GATAATGTTT CATAGTTGGA TATCATAATT TAAACNAGCA AAACCAAATT NAGGGNCAGC
  841 TCATTNCCTC CCACTCATGA TCTANAGATC TATAGATCTC TCGNGGGATC ATTGTTTTTC
  901 TCTTGNNNNC NNNTTTGNGG NNCTNANTAC TGNGGNTTTN CANNGNGTCA GTTTNNNANC
961 CNGNANNAAC NAGNTNANNN NNCCTNNNNN NCNNNNNNNN NN
//
Primer: SV40polyA_R2 (Pr0623)
Region: insert mid(mClover 3')
Sequence file: RDB13934_A6B8i.seq check
>13934_13934_B3C8F_SV40polyA_R2_D01_04_ABI08.ab1
    1 NNNNNNNNNN NNNGNNNNNA AANNNGNNNN NNNCNACNAT TGCATTCATT TTATGTTTCA
   61 GGTTCAGGGG GAGGTGTGGG AGGTTTTTTA AAGCAAGTAA AACCTCTACA AATGTGGTAT
  121 GGCTGATTAT GATCAGTTAT CTAGATTACT TGTACAGCTC GTCCATGCCA TGTGTAATCC
  181 CGGCGGCGGT CACGAACTCC AGCAGGACCA TGTGATCGCG CTTCTCGTTG GGGTCTTTGC
  241 TCAGCTTGGA CTGATGGCTC AGGTAGTGGT TGTCGGGCAG CAGCACGGGG CCGTCGCCGA
  301 TGGGGGTGTT CTGCTGGTAG TGGTCGGCGA GCTGCACGCT GCCGTCCTCA ACGTTGTGGC
  361 GGATCTTGAA GTTAGCCTTG ATGCCGTTCT TCTGCTTGTC GGCCGTGATA TAGACGTTGT
  421 GGCTGTTGAA GTTGTACTCC AGCTTGTGCC CCAGGATGTT GCCGTCCTCC TTGAAGTCGA
  481 TGCCCTTCAG CTCGATGCGG TTCACCAGGG TGTCGCCCTC GAACTTCACC TCGGCGCGGG
  541 TCTTGTAGGT ACCGTCGTCC TTGAAAGAGA TGGTGCGCTC CTGGACGTAG CCTTCGGGCA
  601 TGGCGGACTT GAAGAAGTCG TGCTGCTTCA TGTGGTCGGG GTAGCGGCTG AAGCAGGCCA
  661 CGCCGTANCC GAAGGTGGTC ACGAGGGTGG GCCAGGGCAC GGGCAGCTTG CCGGTGGTGC
  721 AGATGAACTT CANGGTCAGC TTGCCGTTGG TGGCATCGCC CTCGCCCTCG CCGCGGANGC
  781 TGAACTTGTG GCCGTTTACG TCGCCNTCCA GCTCGACCAG NANGGGCACC ACCNCGGTGA
  841 ACAGCTCCTC GCCCTTGCTC ACCATGCTAG CTTTATACNT CCTNNNATCN ATTTTCCNNN
  901 AGNACNGNGC TNCNNCCATT GATANCTTCA CGANNGCCGC CNCNNCCGGN NNNNNGCTTG
961 NNTTTTGNCN NNAANNNNNN NCNNNCNTNN NNNNNTGATN NNNNNNNNN
//

Please visit Sequencing and PCR primers for primer information.


References

Original, user report and related articles

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. [link to RRC of NBRP]
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.