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pMK277 (mClover-NeoR) (#RDB13920)

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

Alternative name mClover-NeoR,1001
Clone info. A plasmid for construction of C-terminal 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.
Vector backbone pBluescriptII KS- (Plasmid)
Size of vector backbone 3.0 kb
Selectable markers Am^r
Growth conditions LB+Amp, 37oC
Gene/insert name Aequorea victoria GFP cDNA
Depositor|Developer Kanemaki, Masato |
 

Distribution information

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Please visit Information of Request for Distribution.[link] 
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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必要書類
提供依頼書  [依頼書の記入例 ]
提供同意書 (MTA, 非営利学術目的用)[Word] [MTAの記入例 ]
提供同意書 (MTA, 営利目的用)[Word]
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。

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB13920 pMK277 (mClover-NeoR) DNA solution JPY 9,460 (not-for-profit academic purpose)
JPY 18,920 (for-profit-research purpose)
plus cost of shipping containers, dry ice (if required) and shipping charge

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 pMK277 (mClover-NeoR) was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13920).

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 RDB13920_A6B8p1_v2.pdf check

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

Primer: M13_-40
Sequence file: RDB13920_A6B8a_2.seq check
>D03070A04_A6B8_1_M13(-40)_A02_02.ab1
    1 AAGCAGGACG CGCGTATACG ACTCACTATA GGGCGAATTG GAGCTCGGAT CCGGTGCAGG
   61 CGCCGCTAGC ATGGTGAGCA AGGGCGAGGA GCTGTTCACC GGGGTGGTGC CCATCCTGGT
  121 CGAGCTGGAC GGCGACGTAA ACGGCCACAA GTTCAGCGTC CGCGGCGAGG GCGAGGGCGA
  181 TGCCACCAAC GGCAAGCTGA CCCTGAAGTT CATCTGCACC ACCGGCAAGC TGCCCGTGCC
  241 CTGGCCCACC CTCGTGACCA CCTTCGGCTA CGGCGTGGCC TGCTTCAGCC GCTACCCCGA
  301 CCACATGAAG CAGCACGACT TCTTCAAGTC CGCCATGCCC GAAGGCTACG TCCAGGAGCG
  361 CACCATCTCT TTCAAGGACG ACGGTACCTA CAAGACCCGC GCCGAGGTGA AGTTCGAGGG
  421 CGACACCCTG GTGAACCGCA TCGAGCTGAA GGGCATCGAC TTCAAGGAGG ACGGCAACAT
  481 CCTGGGGCAC AAGCTGGAGT ACAACTTCAA CAGCCACAAC GTCTATATCA CGGCCGACAA
  541 GCAGAAGAAC GGCATCAAGG CTAACTTCAA GATCCGCCAC AACGTTGAGG ACGGCAGCGT
  601 GCAGCTCGCC GACCACTACC AGCAGAACAC CCCCATCGGC GACGGCCCCG TGCTGCTGCC
  661 CGACAACCAC TACCTGAGCC ATCAGTCCAA GCTGAGCAAA GACCCCAACG AGAAGCGCGA
  721 TCACATGGTC CTGCTGGAGT TCGTGACCGC CGCCGGGATT ACACATGGCA TGGACGAGCT
  781 GTACAAGTAA TCTAGATAAC TGATCATAAT CAGCCATACC ACATTTGTAG AGGTTTTACT
  841 TGCTTTAAAA AACCTCCCAC ACCTCCCCCT GAACCTGAAA CATAAAATGA ATGCAATTGT
  901 TGTTGTTAAC TTGTTTATTG CAGCTTATAA TGGTTACAAA TAAAGCAATA GCATCACAAA
  961 TTTCACAAAT AAAGGCATTT TTTTTCACTG CATTCTAGTT GTGGTTTGTC CAACTCATCC
 1021 ATGTATCTAC GCGTAAATGT AAGCGTCATA TTTGTAAATC CCGTTAACTT GTAATTCAGC
 1081 TCATTTTTAC CAATAGCCGA ATCGCAATTC CCGTTATTAA ATCCAAAGAA CTATGGACAG
//
Primer: SV40pro_F_V2
Sequence file: RDB13920_A6B8b_2.seq check
>D03070A04_A6B8_1_SV40pro_F_V2_G09_21.ab1
    1 GTGGGCTTGT GGAGCTAGGC TTTTGCAAGA TCGATCAAGA GACAGGATGA GGATCGTTTC
   61 GCATGATTGA ACAAGATGGA TTGCACGCAG GTTCTCCGGC CGCTTGGGTG GAGAGGCTAT
  121 TCGGCTATGA CTGGGCACAA CAGACAATCG GCTGCTCTGA TGCCGCCGTG TTCCGGCTGT
  181 CAGCGCAGGG GCGCCCGGTT CTTTTTGTCA AGACCGACCT GTCCGGTGCC CTGAATGAAC
  241 TGCAAGACGA GGCAGCGCGG CTATCGTGGC TGGCCACGAC GGGCGTTCCT TGCGCAGCTG
  301 TGCTCGACGT TGTCACTGAA GCGGGAAGGG ACTGGCTGCT ATTGGGCGAA GTGCCGGGGC
  361 AGGATCTCCT GTCATCTCAC CTTGCTCCTG CCGAGAAAGT ATCCATCATG GCTGATGCAA
  421 TGCGGCGGCT GCATACGCTT GATCCGGCTA CCTGCCCATT CGACCACCAA GCGAAACATC
  481 GCATCGAGCG AGCACGTACT CGGATGGAAG CCGGTCTTGT CGATCAGGAT GATCTGGACG
  541 AAGAGCATCA GGGGCTCGCG CCAGCCGAAC TGTTCGCCAG GCTCAAGGCG AGCATGCCCG
  601 ACGGCGAGGA TCTCGTCGTG ACCCATGGCG ATGCCTGCTT GCCGAATATC ATGGTGGAAA
  661 ATGGCCGCTT TTCTGGATTC ATCGACTGTG GCCGGCTGGG TGTGGCGGAC CGCTATCAGG
  721 ACATAGCGTT GGCTACCCGT GATATTGCTG AAGAGCTTGG CGGCGAATGG GCTGACCGCT
  781 TCCTCGTGCT TTACGGTATC GCCGCTCCCG ATTCGCAGCG CATCGCCTTT CTATCGCCTT
  841 CTTGACGAGT TCTTCTGAGC GGGACTCTGG GGTTTCGAAA TGACCGACCA AGCGACGCCC
  901 AACCTGCCAT CACGAGATTT CGATTCACGC GCTTCTATGA AAGTGGGCTT CGATCGTTTT
  961 CGGACGCGCT GAATGATCTT CAGCGCGGGG ATCTCATGCT GGAAGTCCTC GCCAACCTAA
1021 GGAGATAACT GACACGGAGG AACAATACGT ATGCATCGGC TATGACGGCC ATT
//
Primer: M13R
Sequence file: RDB13920_A6B8c_2.seq check
>D03070A04_A6B8_1_M13R_A01_01.ab1
    1 GTCGGCATTA CCTCACTAAA GGGAACAAAA GCTGGGTACC GGGCCCCCCC TCGAGGGATC
   61 CAACGACCCA ACACCGTGCG TTTTATTCTG TCTTTTTATT GCCGTCATAG CGCGGGTTCC
  121 TTCCGGTATT GTCTCCTTCC GTGTTTCAGT TAGCCTCCCC CTAGGGTGGG CGAAGAACTC
  181 CAGCATGAGA TCCCCGCGCT GGAGGATCAT CCAGCCGGCG TCCCGGAAAA CGATTCCGAA
  241 GCCCAACCTT TCATAGAAGG CGGCGGTGGA ATCGAAATCT CGTGATGGCA GGTTGGGCGT
  301 CGCTTGGTCG GTCATTTCGA ACCCCAGAGT CCCGCTCAGA AGAACTCGTC AAGAAGGCGA
  361 TAGAAGGCGA TGCGCTGCGA ATCGGGAGCG GCGATACCGT AAAGCACGAG GAAGCGGTCA
  421 GCCCATTCGC CGCCAAGCTC TTCAGCAATA TCACGGGTAG CCAACGCTAT GTCCTGATAG
  481 CGGTCCGCCA CACCCAGCCG GCCACAGTCG ATGAATCCAG AAAAGCGGCC ATTTTCCACC
  541 ATGATATTCG GCAAGCAGGC ATCGCCATGG GTCACGACGA GATCCTCGCC GTCGGGCATG
  601 CTCGCCTTGA GCCTGGCGAA CAGTTCGGCT GGCGCGAGCC CCTGATGCTC TTCGTCCAGA
  661 TCATCCTGAT CGACAAGACC GGCTTCCATC CGAGTACGTG CTCGCTCGAT GCGATGTTTC
  721 GCTTGGTGGT CGAATGGGCA GGTAGCCGGA TCAAGCGTAT GCAGCCGCCG CATTGCATCA
  781 GCCATGATGG ATACTTTCTC GGCAGGAGCA AGGTGAGATG ACAGGAGATC CTGCCCCGGC
  841 ACTTCGCCCA ATAGCAGCCA GTCCCTTCCC GCTTCAGTGA CAACGTCGAG CACAGCTGCG
  901 CAAGGAACGC CCGTCGTGGC CAGCCACGAT AGCCGCGCTG CCTCGTCTTG CAGTTCATTC
  961 AGGGCACCGG GACAGGTCGG GTCTTGACAA AAAGAACCCG GGGCGCCCCT GCGCTGACAG
 1021 CCGGAACACG CGGGCATCAG AGCAGCGAAT TGTCTGTTTG TGCCCAGTTC ATAGCGGATA
 1081 GCCTCTCCAC CCAAGCGCCG AGACCTGGCG TGCATCAATC TTGTTCAATT CCAATGGCGA
1141 TAG
//

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 Tungadi, E.A., Human microcephaly ASPM protein is a spindle pole-focusing factor that functions redundantly with CDK5RAP2. J. Cell Sci. 130 (21): 3676-3684 (2017). PMID 28883092.