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pMK281 (mCherry2-Hygro) (#RDB13924)

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

Alternative name mCherry2-Hygro, 975
Clone info. A plasmid for construction of C-terminal 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.
Vector backbone pBluescriptII KS- (Plasmid)
Size of vector backbone 3.0 kb
Selectable markers Am^r
Gene/insert name 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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RDB13924 pMK281 (mCherry2-Hygro) 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 pMK281 (mCherry2-Hygro) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13924).

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 RDB13924_A6B8p1-3.pdf check

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

Primer: M13_-40 (Pr0410)
Region: T7 pro,insert 5'(Linker,mCherry2 5')
Sequence file: RDB13924_A6B8e.seq check
>D03070A08_A6B8_1_M13(-40)_E02_14.ab1
    1 NNNNNNNNNN NNNCNNNNNN GNNTNNCNAT AGGGCGAATT GGANCTCGGA TCCGGTGCAG
   61 GCGCCGCTAG CATGGTGAGC AAGGGCGAGG AGGATAACAT GGCCATCATC AAGGAGTTCA
  121 TGCGCTTCAA GGTGCACATG GAGGGCTCCG TGAACGGCCA CGAGTTCGAG ATCGAGGGCG
  181 AGGGCGAGGG CCGCCCCTAC GAGGGCACCC AGACCGCCAA GCTGAAGGTG ACCAAGGGTG
  241 GCCCCCTGCC CTTCGCCTGG GACATCCTGT CCCCTCAGTT CATGTACGGC TCCAAGGCCT
  301 ACGTGAAGCA CCCCGCCGAC ATCCCCGACT ACTTGAAGCT GTCCTTCCCC GAGGGCTTCA
  361 ATTGGGAGCG CGTGATGAAC TTCGAGGACG GCGGCGTGGT GACCGTGACC CAGGACTCCT
  421 CCCTGCAGGA CGGCGAGTTC ATCTACAAGG TGAAGCTGCG CGGCACCAAC TTCCCCTCCG
  481 ACGGCCCCGT AATGCAGTGT CGTACCATGG GCTGGGAGGC CTCCACTGAG CGGATGTACC
  541 CCGAGGACGG CGCCCTGAAG GGCGAGATCA AGCAGAGGCT GAAGCTGAAG GACGGCGGCC
  601 ACTACGACGC TGAGGTCAAG ACCACCTACA AGGCCAAGAA GCCCGTGCAG CTGCCCGGCG
  661 CCTACAACGT CGACATCAAG TTGGACATCC TTTCCCACAA CGAGGNCTAC ACCATCGTGG
  721 AACAGTACGA ACGCGCCGAG GGCCGCCACT CCACCGGCGG CATGGACGAN CTGTACAAGT
  781 AATCTAGATA ACTGATCATA ATCAGCCATA CCACATTTGT AGAGGTTTTA CTTGCTTTNA
  841 AAAANCTCCC ACACCNCCCC CNGANNCTGA AACATAAAAN GAATGCNATT GTTGTTGNTA
  901 ACTTGTTNAT TGCAGCTTNN AATGNTTACN AATAAANCAA TAGCNTCNCN ANNTTCACAA
  961 ATAAAGCATT TTTTTCNCTG NNNTCTANNN GNGGNTNGNN CNAANNCATN NNNGTATNNN
1021 NNNNNNN
//
Primer: M13R (Pr0160)
Region: insert 3'(loxP,mPGK ter,HygR)
Sequence file: RDB13924_A6B8f.seq check
>D03070A08_A6B8_1_M13R_E01_13.ab1
    1 NNNNNNNNNN NNNNNNNNNN NANGNNNNNN AAGCTGGGTA NCGGATCCTA GTGAACCTCT
   61 TCGAGGGACC TAATAACTTC GTATAGCATA CATTATACGA AGTTATATTA AGGGTTCCGG
  121 ATCTCGACCA GCTTCTGATG GAATTAGAAC TTGGCAAAAC AATACTGAGA ATGAAGTGTA
  181 TGTGGAACAG AGGCTGCTGA TCTCGTTCTT CAGGCTATGA AACTGACACA TTTGGAAACC
  241 ACAGTACTTA GAACCACAAA GTGGGAATCA AGAGAAAAAC AATGATCCCA CGAGAGATCT
  301 ATAGATCTAT AGATCATGAG TGGGAGGAAT GAGCTGGCCC TTAATTTGGT TTTGCTTGTT
  361 TAAATTATGA TATCCAACTA TGAAACATTA TCATAAAGCA ATAGTAAAGA GCCTTCAGTA
  421 AAGAGCAGGC ATTTATCTAA TCCCACCCCA CCCCCACCCC CGTAGCTCCA ATCCTTCCAT
  481 TCAAAATGTA GGTACTCTGT TCTCACCCTT CTTAACAAAG TATGACAGGA AAAACTTCCA
  541 TTTTAGTGGA CATCTTTATT GTTTAATAGA TCATCAATTT CTGCATCCCG GGGATCTGAT
  601 ATCATCGATG CATGGGGTCG TGCGCTCCTT TCGGTCGGGC GCTGCGGGTC GTGGGGCGGG
  661 CGGCTAGCCT ATTCCTTTGC CCTCGGACGA GTGCTGGGGC GTCGGTTTCC ACTATCGGCG
  721 AGTACTTCTA CACAGCCATC GGTCCAGACN GCCGCGCTTC TGCGGGCGAT TTGTGTACGC
  781 CCGACAGTCC CGGCTCCNGA TCGGANGATT GCGTCGCATC GACCCTGCGC CCAAGCTGNN
  841 NTCATCGAAN NTGCCGTCNA CCAANNCTCT GANAGAGTTG GTCNNGANCA ATGCNGNANC
  901 ATATACGCCC NGNANNCNCN GNNATCCTGC NANCNNNNGA TGNCNNNCNC TCNNAANNNN
  961 NNCGTCTGNN GCNNCNNNNN ANNNCNNANC NNNNGNNCNN NCNNNNNNNA NNNNGNNNNN
1021 NNNNN
//
Primer: PGK_N_rev (Pr0243)
Region: insert mid(loxP,SV40 pA,mCherry2 3')
Sequence file: RDB13924_A6B8g.seq check
>D03070A08_A6B8_1_PGK-N-rev_C06_09.ab1
    1 NNNNNNNNNN NNNNNNNNNC NGGNANANTT CGAGGTCGAG ATCCTAAGCT TGGCTGGACG
   61 TAAACTCCTC TTCAGACCTA ATAACTTCGT ATAGCATACA TTATACGAAG TTATATTAAG
  121 GGTTATTGAA TATGATCGAC GCGTTAAGAT ACATTGATGA GTTTGGACAA ACCACAACTA
  181 GAATGCAGTG AAAAAAATGC TTTATTTGTG AAATTTGTGA TGCTATTGCT TTATTTGTAA
  241 CCATTATAAG CTGCAATAAA CAAGTTAACA ACAACAATTG CATTCATTTT ATGTTTCAGG
  301 TTCAGGGGGA GGTGTGGGAG GTTTTTTAAA GCAAGTAAAA CCTCTACAAA TGTGGTATGG
  361 CTGATTATGA TCAGTTATCT AGATTACTTG TACAGCTCGT CCATGCCGCC GGTGGAGTGG
  421 CGGCCCTCGG CGCGTTCGTA CTGTTCCACG ATGGTGTAGT CCTCGTTGTG GGAAAGGATG
  481 TCCAACTTGA TGTCGACGTT GTAGGCGCCG GGCAGCTGCA CGGGCTTCTT GGCCTTGTAG
  541 GTGGTCTTGA CCTCAGCGTC GTAGTGGCCG CCGTCCTTCA GCTTCAGCCT CTGCTTGATC
  601 TCGCCCTTCA GGGCGCCGTC CTCGGGNNNN NNCCGCTCAG TGGAGGCCTC CCAGCCCATG
  661 GTACGACACT GCATTACGGG GCCGTCGGAN GGGAAGTTGG TGCCNCGCAN CTTCACCTTG
  721 TAGATGAACT CNNCGTCCTG CANGGANGAN NNNNTGGGTC ACGGTCACCN NNNCNCCGTC
  781 CTCNAAGTTC NTCNCGCGCT CCCNNNNNNN NNCCCTCNGG GGAAGGACAG CTTCANNNNN
841 NNNNN
//
Primer: PGKpro_F (Pr0249)
Region: insert mid(HygR)
Sequence file: RDB13924_A6B8h.seq check
>D03070A08_A6B8_1_PGK-proF_C07_07.ab1
    1 NNNNNNNNNN NNNNNNNNNN NTCTCCNGGC CTTTCGACCT GCATCCNTCT 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 TCTGGANGCC GTGGTTGGCT TGTATGGAGC AGCAGANGCG CTACTTCGAG
  841 CGGANGCATC CGGANCTTGC NNGATCGCCG CGGCNNCNGG CGTATATGCT CCGCATTGGT
  901 CTTGACNNAC TCTATCANAN CTNNNNTGAN NGCNATTTCN ATGANGCAGC TNNNNNCNNG
  961 NTCGATGCNA CNNNNNCGTC CNATNNNGNA NNCNNGNNNN GNCNGGNNGT ACNNNANNNN
1021 NNNNNN
//

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 Natsume, T., Acute inactivation of the replicative helicase in human cells triggers MCM8-9-dependent DNA synthesis. Genes Dev. (2017). PMID 28487407.