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

pMK292 (mAID-mCherry2-NeoR) (#RDB13935)

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

Alternative name mAID-mCherry2-NeoR, 971
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. Please note that this plasmid does not contain the loxP sites, which are intended for removal of the marker.
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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RDB13935 pMK292 (mAID-mCherry2-NeoR) 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 pMK292 (mAID-mCherry2-NeoR) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13935).

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 RDB13935_A6B8p1-4.pdf check

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

Primer: M13_-40 (Pr0410)
Region: insert 5'(Linker,mAID,mCherry2 5')
Sequence file: RDB13935_A6B8i.seq check
>D03070A19_A6B8_1_M13(-40)_D05_11.ab1
    1 NNNNNNNNNN NNNNNNNNCN NNNNNNGANT NNNNNTAGGG CGAANTGGAG CTCCCCGGGN
   61 TCCGGTGCAG GCGCCAAGGA GAAGAGTGCT TGTCCTAAAG ATCCAGCCAA ACCTCCGGCC
  121 AAGGCACAAG TTGTGGGATG GCCACCGGTG AGATCATACC GGAAGAACGT GATGGTTTCC
  181 TGCCAAAAAT CAAGCGGTGG CCCGGAGGCG GCGGCGTTCG TGAAGGTATC AATGGACGGA
  241 GCACCGTACT TGAGGAAAAT CGATTTGAGG ATGTATAAAG CTAGCATGGT GAGCAAGGGC
  301 GAGGAGGATA ACATGGCCAT CATCAAGGAG TTCATGCGCT TCAAGGTGCA CATGGAGGGC
  361 TCCGTGAACG GCCACGAGTT CGAGATCGAG GGCGAGGGCG AGGGCCGCCC CTACGAGGGC
  421 ACCCAGACCG CCAAGCTGAA GGTGACCAAG GGTGGCCCCC TGCCCTTCGC CTGGGACATC
  481 CTGTCCCCTC AGTTCATGTA CGGCTCCAAG GCCTACGTGA AGCACCCCGC CGACATCCCC
  541 GACTACTTGA AGCTGTCCTT CCCCGAGGGC TTCAATTGGG AGCGCGTGAT GAACTTCGAG
  601 GACGGCGGCG TGGTGACCGT GACNNNNNAC TCCTCCCTGC AGGACGGCGA GTTCATCTAC
  661 AAGGTGAAGC TGCGCGGCAC CNACTTCCCC TCCGACGGCC CCGTAATGCA GTGTCGTACC
  721 ATGGGCTGGG AGGCCTCCAC TGNNNNGATG TNNNCCCGAG GACGGCGCCC TGAANGGCGA
  781 GATCNAGCAG ANGCTGAANG CTGANNGACG GCGGCCACTA CNACGCTGAN GTCAAGACCA
  841 CCTANNNNGC CNAGAAGCCC GTGCAGCTGN CCNGCGCNNN NNACGTCNAN NNCNNGTTNG
  901 GANATCCTTN NNNNNNNCNN GGACTANNNC NTCGNGGAAN NANNNNNNAA NNNNNNNNNG
  961 GGNCGNCNNC TNNNNCNGNN GGCNNNGNNN NNNCNGTNCA NNNNNTCTNN NANNANCTGA
 1021 NNNNNNANNC NNNNNTNNCA CATTTNNNNN NANGGNTTTT ANNNNNNNTT TNNNNNNNNN
1081 N
//
Primer: M13R (Pr0160)
Region: insert 3'(NeoR_KanR 3')
Sequence file: RDB13935_A6B8j.seq check
>D03070A19_A6B8_1_M13R_D04_10.ab1
    1 NNNNNNNNNN NNNNNNNNNN ANGGGNNNNN AAGCTGGGTA NCGGATCCNA CGACCCAACA
   61 CCGTGCGTTT TATTCTGTCT TTTTATTGCC GTCATAGCGC GGGTTCCTTC CGGTATTGTC
  121 TCCTTCCGTG TTTCAGTTAG CCTCCCCCTA GGGTGGGCGA AGAACTCCAG CATGAGATCC
  181 CCGCGCTGGA GGATCATCCA GCCGGCGTCC CGGAAAACGA TTCCGAAGCC CAACCTTTCA
  241 TAGAAGGCGG CGGTGGAATC GAAATCTCGT GATGGCAGGT TGGGCGTCGC TTGGTCGGTC
  301 ATTTCGAACC CCAGAGTCCC GCTCAGAAGA ACTCGTCAAG AAGGCGATAG AAGGCGATGC
  361 GCTGCGAATC GGGAGCGGCG ATACCGTAAA GCACGAGGAA GCGGTCAGCC CATTCGCCGC
  421 CAAGCTCTTC AGCAATATCA CGGGTAGCCA ACGCTATGTC CTGATAGCGG TCCGCCACAC
  481 CCAGCCGGCC ACAGTCGATG AATCCAGAAA AGCGGCCATT TTCCACCATG ATATTCGGCA
  541 AGCAGGCATC GCCATGGGTC ACGACGAGAT CCTCGCCGTC GGGCATGCTC GCCTTGAGCC
  601 TGGCGAACAG TTCGGCTGGC GCGAGCCCCT GATGCTCTTC GTCCAGATCA TCCTGATCGA
  661 CAAGACCGGC TTCCATCCGA GTACGTGCTC GCTCGATGCG ATGTTTCGCT TGGTGGTCGA
  721 ATGGGCAGGT AGCCGGATCA AGCGTATGCA GCCGCCGCAT TGCATCAGCC ATGATGGATA
  781 CTTTCTCGGC AGGAGCAAGG TGAGATGACA GGAGATCCTG CCCCGGCACT TCGCCCAATA
  841 GCAGCCAGTC CCTTCCCGCT TCAGTGACAA CGTCNAGCAC AGCTGCNCAN NGAACGCCCG
  901 NCGNGGCCAG CCACGATAGC CGCGCNGCCN CGTCTNGCAG TTCANTCNNN GCANCNGNNN
961 GGTCNGTCTN GANNAAAANA NNCNGGCGCC CNNNCNNNNN NNNNCNNNNN NNNNN
//
Primer: SV40polyA_R2 (Pr0623)
Region: insert mid(SV40 ter,mCherry2 3')
Sequence file: RDB13935_A6B8k.seq check
>13935_13935_A9KsF_SV40polyA_R2_F09_18_ABI24.ab1
    1 NNNNNNNNGN ANNANNNNGT TAACAACAAC AATTGCATTC ATTTTATGTT TCNGGTTCAG
   61 GGGGAGGTGT GGGAGGTTTT TTAAAGCAAG TAAAACCTCT ACAAATGTGG TATGGCTGAT
  121 TATGATCAGT TATCTAGATT ACTTGTACAG CTCGTCCATG CCGCCGGTGG AGTGGCGGCC
  181 CTCGGCGCGT TCGTACTGTT CCACGATGGT GTAGTCCTCG TTGTGGGAAA GGATGTCCAA
  241 CTTGATGTCG ACGTTGTAGG CGCCGGGCAG CTGCACGGGC TTCTTGGCCT TGTAGGTGGT
  301 CTTGACCTCA GCGTCGTAGT GGCCGCCGTC CTTCAGCTTC AGCCTCTGCT TGATCTCGCC
  361 CTTCAGGGCG CCGTCCTCGG GGTACATCCG CTCAGTGGAG GCCTCCCAGC CCATGGTACG
  421 ACACTGCATT ACGGGGCCGT CGGAGGGGAA GTTGGTGCCG CGCAGCTTCA CCTTGTAGAT
  481 GAACTCGCCG TCCTGCAGGG AGGAGTCCTG GGTCACGGTC ACCACGCCGC CGTCCTCGAA
  541 GTTCATCACG CGCTCCCAAT TGAAGCCCTC GGGGAAGGAC AGCTTCAAGT AGTCGGGGAT
  601 GTCGGCGGGG TGCTTCACGT AGGCCTTGGA GCCGTACATG AACTGAGGGG ACAGGATGTC
  661 CCAGGCGAAG GGCAGGGGGC CACCCTTGGT CACCTTCAGC TTGNCGGTCT GGGTGCCCTC
  721 GTANGGGCGG NCCTCGCCCT CGCCCTCGAT CTCGAACTCG TGGCCGTTCA CGGAGCCCTC
  781 CATGTGCANC TTGAAGCGCA TGAACTCCTT GATGATGGCC ATGTTATCCT CCTCGCCCTT
  841 GCTCACCATG CTAGCTTTAT ACATCCTCAA NTCGATTTTC CTCNAGTACN GNGCNNCGNN
  901 NATTGATNNC TTCACNAACG CCGCCGCNNN CNGGNCACCG CTNGANTTTT GGNNNNAANN
961 NNTCNCGTTC NNNCGGTATG ATNNNNNNNN NN
//
Primer: SV40pro_F_V2 (Pr0154)
Region: insert mid(NeoR_KanR 5 ')
Sequence file: RDB13935_A6B8l.seq check
>D03070A19_A6B8_1_SV40pro_F_V2_F10_16.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNGCTTTTG CAAGNNCGAT CNAGAGANNG GATGAGGNTC
   61 GTTTCGCATG ATTGAACNNG ATGGATTGCA CGCAGGTTCT CCGGCCGCTT GGGTGGAGAG
  121 GCTATTCGGC TATGACTGGG CACAACAGAC AATCGGCTGC TCTGATGCCG CCGTGTTCCG
  181 GCTGTCAGCG CAGGGGCGCC CGGTTCTTTT TGTCAAGACC GACCTGTCCG GTGCCCTGAA
  241 TGAACTGCAA GACGAGGCAG CGCGGCTATC GTGGCTGGCC ACGACGGGCG TTCCTTGCGC
  301 AGCTGTGCTC GACGTTGTCA CTGAAGCGGG AAGGGACTGG CTGCTATTGG GCGAAGTGCC
  361 GGGGCAGGAT CTCCTGTCAT CTCACCTTGC TCCTGCCGAG AAAGTATCCA TCATGGCTGA
  421 TGCAATGCGG CGGCTGCATA CGCTTGATCC GGCTACCTGC CCATTCGACC ACCAAGCGAA
  481 ACATCGCATC GAGCGAGCAC GTACTCGGAT GGAAGCCGGT CTTGTCGATC AGGATGATCT
  541 GGACGAAGAG CATCAGGGGC TCGCGCCAGC CGAACTGTTC GCCAGGCTCA AGGCGAGCAT
  601 GCCCGACGGC GAGGATCTCG TCGTGACCCA TGGCGATGCC TGCTTGCCGA ATATCATGGT
  661 GGAAAATGGC CGCTTTTCTG GATTCATCGA CTGTGGCCGG CTGGGTGTGG CGGACCGCTA
  721 TCAGGACATA GCGTTGGCTA CCCGTGATAT TGCTGANNAG CTTGGCGGCG AATGGGCTGA
  781 CCGCTTCCTC GTGCTTTACG GTATCGCCGC TNCCGATTCG CAGCGCATCG CCTTCTATCG
  841 CCTTCTTGAC GANTTCTTCT GAGCGGGNNT CTGGGGTTCG AANTGACCGA NCAAGCGNNG
  901 CCCNANCTGN CNTCNCGANN NTNCNATTCC NCCGCCGCCT TCTATGAANN NNNGGGGCTT
961 NNNNNNNNNT TTNCNNGNNN NNNNGNTNNN NGNNNNNNNN NN
//

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 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 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.