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

pMK294 (mAID-mCherry2-Bsr) (#RDB13937)

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

Alternative name mAID-mCherry2-Bsr, 976
Clone info. A plasmid for construction of C-terminal mAID-mCherry 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 Discosoma sp. RFP 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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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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RDB13937 pMK294 (mAID-mCherry2-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 pMK294 (mAID-mCherry2-Bsr) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13937).

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 RDB13937_A6B8p1-3.pdf check

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

Primer: M13_-40 (Pr0410)
Region: insert mid(Linker,mAID,mCherry2 5')
Sequence file: RDB13937_A6B8e.seq check
>D03070A21_A6B8_1_M13(-40)_F05_17.ab1
    1 NNNNNNNNNN NNNCNNNNNN GACTNNCNAT AGGGCGAATT GGAGCTCCCC GGGNTCCGGT
   61 GCAGGCGCCA AGGAGAAGAG TGCTTGTCCT AAAGATCCAG CCAAACCTCC GGCCAAGGCA
  121 CAAGTTGTGG GATGGCCACC GGTGAGATCA TACCGGAAGA ACGTGATGGT TTCCTGCCAA
  181 AAATCAAGCG GTGGCCCGGA GGCGGCGGCG TTCGTGAAGG TATCAATGGA CGGAGCACCG
  241 TACTTGAGGA AAATCGATTT GAGGATGTAT AAAGCTAGCA TGGTGAGCAA GGGCGAGGAG
  301 GATAACATGG CCATCATCAA GGAGTTCATG CGCTTCAAGG TGCACATGGA GGGCTCCGTG
  361 AACGGCCACG AGTTCGAGAT CGAGGGCGAG GGCGAGGGCC GCCCCTACGA GGGCACCCAG
  421 ACCGCCAAGC TGAAGGTGAC CAAGGGTGGC CCCCTGCCCT TCGCCTGGGA CATCCTGTCC
  481 CCTCAGTTCA TGTACGGCTC CAAGGCCTAC GTGAAGCACC CCGCCGACAT CCCCGACTAC
  541 TTGAAGCTGT CCTTCCCCGA GGGCTTCAAT TGGGAGCGCG TGATGAACTT CGAGGACGGC
  601 GGCGTGGTGA CCGTGACCCA GGACTCCTCC CTGCAGGACG GCGAGTTCAT CTACAAGGTG
  661 AAGCTGCGCG GCACNNAACT TCCCCTCCGA CGGCCCCGTA ATGNAGTGTC GTACCATGGG
  721 CTGGGANGNC TCCACTGANC GGATGTACCN CGANGANGGC GCCCTGAANG GNGAGNTCAA
  781 NCANANGCTG AAGCTGANNN CGGCGGCCNC TACNANGCTG ANGTCNAGAC CACCTACNAN
  841 GNNNNAGAAG CCCGTGCANC TGCCCNGNGC CNACAANNNC NANNTCANNT TGGANNNCCT
  901 TTNNCCANNN NNNNNACNNN NCCNNNGNNN AACANTNCNA ACGCNNCNNN NGGGNCNNCN
  961 NNTCCNNCNN NNNGNNNNGN NNNNNNCNGT NNNNANNTAN NNNNANANNN NNGNNNNNNN
1021 ANNNNNNNNN CNNNNNNNNN N
//
Primer: M13R (Pr0160)
Region: insert 3'(loxP,mPGK ter,bsr)
Sequence file: RDB13937_A6B8f.seq check
>D03070A21_A6B8_1_M13R_F04_16.ab1
    1 NNNNNNNNNN NNNNNNNNNN NAANGGGNNN NAAAGCTGGG 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 CTTACNNCTC GAATACTTCT ATCTACTTCG TCAGANNANN
  841 NTGTCNNACA GCTACNATCG TGTCNAANTC CTTTTGTNNN TTCNNAACTG CNCNANCNAT
901 CNCAANGGCN NCTGCNCAAA CNGTTANTCN NNCNNNNNNN NNN
//
Primer: PGK_N_rev (Pr0243)
Region: insert mid(loxP,SV40 pA,mCherry2 3')
Sequence file: RDB13937_A6B8g.seq check
>D03070A21_A6B8_1_PGK-N-rev_H06_24.ab1
    1 NNNNNNNNNN NNNNNNNGNN NANTTCGAGG TCGAGNTCCT AAGCTTGGCT GGACGTAAAC
   61 TCCTCTTCNG ACCTAATAAC TTCGTATAGC ATACATTATA CGAAGTTATA TTAAGGGTTA
  121 TTGAATATGA TCGACGCGTT AAGATACATT GATGAGTTTG GACAAACCAC AACTAGAATG
  181 CAGTGAAAAA AATGCTTTAT TTGTGAAATT TGTGATGCTA TTGCTTTATT TGTAACCATT
  241 ATAAGCTGCA ATAAACAAGT TAACAACAAC AATTGCATTC ATTTTATGTT TCAGGTTCAG
  301 GGGGAGGTGT GGGAGGTTTT TTAAAGCAAG TAAAACCTCT ACAAATGTGG TATGGCTGAT
  361 TATGATCAGT TATCTAGATT ACTTGTACAG CTCGTCCATG CCGCCGGTGG AGTGGCGGCC
  421 CTCGGCGCGT TCGTACTGTT CCACGATGGT GTAGTCCTCG TTGTGGGAAA GGATGTCCAA
  481 CTTGATGTCG ACGTTGTAGG CGCCGGGCAG CTGCACGGGC TTCTTGGCCT TGTAGGTGGT
  541 CTTGACCTCA GCGTCGTAGT GGCCGCCGTC CTTCAGCTTC AGCCTCTGCT TGATCTCGCC
  601 CTTCAGGGCG CCGTCCTCGG GGTACATCCG CTCAGTGGAG GCCTCCCAGC CCATGGTACG
  661 ACACTGCATT ACGGGGCCGT CGGANGGGAA GTTGGTGCCG CGCAGCTTCA CCTTGTAGAT
  721 GAACTCGCCG TCCTGCANGG ANGANTCCTG GGTCACGGTC ACCACGCCGC CNTCCTCGAA
  781 NTTCATCACG CGCTCCCAAT TGAAGCCCTC GGGGAANGNN AGCTTCAAGT ANTCNGGGNN
  841 GTCNGCGGGG TGCTTCACGT ANGCCTTNNA NCCNTACATG AACTGANGGG NNAGGANGTC
  901 CNAGGCGAAN GGNANGGGGN CACCCTNNGT NNNCNTCANC TTGGCGGNCN GGGNGCCCNC
961 NNNNGGCNGN CNNCGCCCTC NNNNNNNTNN NNNACNNNNN NNNNN
//
Primer: PGKpro_F (Pr0249)
Region: insert mid(bsr,mPGK ter)
Sequence file: RDB13937_A6B8h.seq check
>D03070A21_A6B8_1_PGK-proF_C08_08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NTCTCCNGGC 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 ATTGGAGCTA CGGGGGTGGG
  721 GGTGGGGTGG GATTAGATAA ATGCCTGCTC TTTACTGAAG GCTCTTTACT ATTGCTTTAT
  781 GATAATGNTT CATAGTTGGA TATCATAATT TAAACAAGCN AAACCAAATT NANGGGNCAG
  841 CTCANTCCTC CCACTCATGA TCNATAGATC TATAGATCTC TCGNGGGATC ATTGNTTTTC
  901 TCTTGANTCC CACTTTNNGG NNCTANGTNN TGNGGTTNNC NNNANGNGNN ANTTTCNNAG
961 CNNGNANNAN CNAGATNNNN NNNNCNNNNN NNCNNNNNNN NNN
//
Primer: mCherry-C_R2 (Pr0792)
Region: insert mid(mCherry2 5',mAID 3')
Sequence file: RDB13937_A6B8i.seq check
>13937_13937_B3C8F_mCherry-C_R2_E01_05_ABI08.ab1
    1 NNNNNNNNNN NNNNCNNNNT TNNGCTTNNG CCTCTGCTTG ATCTCGCCCT TCAGGGCGCC
   61 GTCCTCGGGG TACATCCGCT CAGTGGAGGC CTCCCAGCCC ATGGTACGAC ACTGCATTAC
  121 GGGGCCGTCG GAGGGGAAGT TGGTGCCGCG CAGCTTCACC TTGTAGATGA ACTCGCCGTC
  181 CTGCAGGGAG GAGTCCTGGG TCACGGTCAC CACGCCGCCG TCCTCGAAGT TCATCACGCG
  241 CTCCCAATTG AAGCCCTCGG GGAAGGACAG CTTCAAGTAG TCGGGGATGT CGGCGGGGTG
  301 CTTCACGTAG GCCTTGGAGC CGTACATGAA CTGAGGGGAC AGGATGTCCC AGGCGAAGGG
  361 CAGGGGGCCA CCCTTGGTCA CCTTCAGCTT GGCGGTCTGG GTGCCCTCGT AGGGGCGGCC
  421 CTCGCCCTCG CCCTCGATCT CGAACTCGTG GCCGTTCACG GAGCCCTCCA TGTGCACCTT
  481 GAAGCGCATG AACTCCTTGA TGATGGCCAT GTTATCCTCC TCGCCCTTGC TCACCATGCT
  541 AGCTTTATAC ATCCTCAAAT CGATTTTCCT CAAGTACGGT GCTCCGTCCA TTGATACCTT
  601 CACGAACGCC GCCGCCTCCG GGCCACCGCT TGATTTTTGG CAGGAAACCA TCACGTTCTT
  661 CCGGTATGAT CTCACCGGTG GCCATCCCAC AACTTGTGCC TTGGCCGGAG GTTTGGCTGG
  721 ATCTTTANGA CAAGCACTCT TCTCCTTGGC GCCTGCACCG GATCCCGGGG AGCTCCAATT
  781 CGCCCTATAG TGAGTCGTAT TACGCGCGCT CACTGGCCGT CGTTTTACAA CGTCNTGACT
  841 GNGAAAACCC TGGCGTTACC CAACNTAATC GCCTNNCAGC ACATCCCCCT TTCGCCAGCT
  901 NGCGTAATAN CNAANANGCC CGCACCGATN GCCNNTCCNN NNAGTNGCNC ANCCTGNNTG
961 GCNNNNNNNA ANTNGNNNGC GNNNNTATTT TNNNNAANNN NNNNNNN
//

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.