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pMK283 (Stag-3FLAG-Neo)

A plasmid for construction of C-terminal Stag-3FLAG tagging donors.

Catalog number RDB13926
Resource name pMK283 (Stag-3FLAG-Neo)
Alternative name Stag-3FLAG-Neo
Clone info. A plasmid for construction of C-terminal Stag-3FLAG 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.
Sequence analysis is not reliable around MluI site adjacent to SV40 poly(A) site probably because of tangled tertiary structure induced by unexpected palindrome sequences.
Vector backbone pBluescriptII KS- (Plasmid)
Size of vector backbone 3.0 kb
Selectable markers Am^r
Depositor Kanemaki, Masato |

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
Terms and conditions set forth by the DEPOSITOR 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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Catalog # Resource name Shipping form Fee (non-profit org.)
RDB13926 pMK283 (Stag-3FLAG-Neo) DNA solution

cDNA Request [in Japanese] [in English]

References and tips

Electronic file

Featured content

Featured content Auxin Inducible Degron (AID) System (English text)
Featured content Auxin Inducible Degron (AID) System (Japanese text)
Featured content Genome Editing (English text)
Featured content Genome Editing (Japanese text)
Featured content Empty Backbone (English text)
Featured content Empty Backbone (Japanese text)

References

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.

User reports and related articles (go to bottom)


Sequence information

RIKEN BRC has sequenced portions of this material for quality test. Primers and the results are shown below.

Data are summerized on test sheet below.

Test sheet RDB13926_A6B8p1_v2.pdf

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

Primer: M13_-40 RDB13926_A6B8a_2.seq
>D03070A10_A6B8_3_M13(-40)_B01_04.ab1
    1 CAGCCAGCGC GCGCGTATAC GACTCACTAT AGGGCGAATT GGAGCTCGGA TCCGGTGCAG
   61 GCGCCAAGGA AACCGCCGCT GCAAAATTCG AACGCCAGCA CATGGACAGC CGATCCCGGG
  121 CTGACTACAA AGACCATGAC GGTGATTATA AAGATCATGA CATCGACTAC AAGGATGACG
  181 ATGACAAGTA GTGTACAAGT AATCTAGATA ACTGATCATA ATCAGCCATA CCACATTTGT
  241 AGAGGTTTTA CTTGCTTTAA AAAACCTCCC ACACCTCCCC CTGAACCTGA AACATAAAAT
  301 GAATGCAATT GTTGTTGTTA ACTTGTTTAT TGCAGCTTAT AATGGTTACA AATAAAGCAA
  361 TAGCATCACA AATTTCACAA ATAAAGCATT TTTTTCACTG CATTCTAGTT GTGGTTTGTC
421 CAAACTCATC AATGTATCTT AACGCGTAAA TTGTAAGCG
//
Primer: SV40pro_F_V2 RDB13926_A6B8b_2.seq
>D03070A10_A6B8_3_SV40pro_F_V2_H09_24.ab1
    1 GTAGGCTTTT TGGAGGCTAG GCTTTTGCAA GATCGATCAA GAGACAGGAT GAGGATCGTT
   61 TCGCATGATT GAACAAGATG GATTGCACGC AGGTTCTCCG GCCGCTTGGG TGGAGAGGCT
  121 ATTCGGCTAT GACTGGGCAC AACAGACAAT CGGCTGCTCT GATGCCGCCG TGTTCCGGCT
  181 GTCAGCGCAG GGGCGCCCGG TTCTTTTTGT CAAGACCGAC CTGTCCGGTG CCCTGAATGA
  241 ACTGCAAGAC GAGGCAGCGC GGCTATCGTG GCTGGCCACG ACGGGCGTTC CTTGCGCAGC
  301 TGTGCTCGAC GTTGTCACTG AAGCGGGAAG GGACTGGCTG CTATTGGGCG AAGTGCCGGG
  361 GCAGGATCTC CTGTCATCTC ACCTTGCTCC TGCCGAGAAA GTATCCATCA TGGCTGATGC
  421 AATGCGGCGG CTGCATACGC TTGATCCGGC TACCTGCCCA TTCGACCACC AAGCGAAACA
  481 TCGCATCGAG CGAGCACGTA CTCGGATGGA AGCCGGTCTT GTCGATCAGG ATGATCTGGA
  541 CGAAGAGCAT CAGGGGCTCG CGCCAGCCGA ACTGTTCGCC AGGCTCAAGG CGAGCATGCC
  601 CGACGGCGAG GATCTCGTCG TGACCCATGG CGATGCCTGC TTGCCGAATA TCATGGTGGA
  661 AAATGGCCGC TTTTCTGGAT TCATCGACTG TGGCCGGCTG GGTGTGGCGG ACCGCTATCA
  721 GGACATAGCG TTGGCTACCC GTGATATTGC TGAAGAGCTT GGCGGCGAAT GGGCTGACCG
  781 CTTCCTCGTG CTTTACGGTA TCGCCGCTCC CGATTCGCAG CGCATCGCCT TCTATCGCCT
  841 TCTTGACGAG TTCTTCTGAG CGGGACTCTG GGGTTCGAAA TGACCGACCA AGCGACGCCC
  901 AACCTGCCAT CACGAGATTT CGATTCCACC GCCGCCTTCT ATGAAAGGTT GGGCTTCGGA
  961 ATCGTTTTCC CGGGACGCCG GCTGGATGAT CCCTCCAGCG CGGGGGATCT CATGCTGGGA
 1021 GGTTCTTTCG CCCACCCCTA GGGGGAGGCT ACTGAAACAC GGATGAGAAC ATACCGAGAA
1081 CCCGCGCTAT GACGCCAATA AAGACGAGTA AAACGGGACT AACT
//
Primer: M13R RDB13926_A6B8c_2.seq
>D03070A10_A6B8_3_M13R_A01_01.ab1
    1 CTGGGGCACT ACCCTCACTA AAGGGAACAA AAGCTGGGTA CCGGGCCCCC CCTCGAGGGA
   61 TCCAACGACC CAACACCGTG CGTTTTATTC TGTCTTTTTA TTGCCGTCAT AGCGCGGGTT
  121 CCTTCCGGTA TTGTCTCCTT CCGTGTTTCA GTTAGCCTCC CCCTAGGGTG GGCGAAGAAC
  181 TCCAGCATGA GATCCCCGCG CTGGAGGATC ATCCAGCCGG CGTCCCGGAA AACGATTCCG
  241 AAGCCCAACC TTTCATAGAA GGCGGCGGTG GAATCGAAAT CTCGTGATGG CAGGTTGGGC
  301 GTCGCTTGGT CGGTCATTTC GAACCCCAGA GTCCCGCTCA GAAGAACTCG TCAAGAAGGC
  361 GATAGAAGGC GATGCGCTGC GAATCGGGAG CGGCGATACC GTAAAGCACG AGGAAGCGGT
  421 CAGCCCATTC GCCGCCAAGC TCTTCAGCAA TATCACGGGT AGCCAACGCT ATGTCCTGAT
  481 AGCGGTCCGC CACACCCAGC CGGCCACAGT CGATGAATCC AGAAAAGCGG CCATTTTCCA
  541 CCATGATATT CGGCAAGCAG GCATCGCCAT GGGTCACGAC GAGATCCTCG CCGTCGGGCA
  601 TGCTCGCCTT GAGCCTGGCG AACAGTTCGG CTGGCGCGAG CCCCTGATGC TCTTCGTCCA
  661 GATCATCCTG ATCGACAAGA CCGGCTTCCA TCCGAGTACG TGCTCGCTCG ATGCGATGTT
  721 TCGCTTGGTG GTCGAATGGG CAGGTAGCCG GATCAAGCGT ATGCAGCCGC CGCATTGCAT
  781 CAGCCATGAT GGATACTTTC TCGGCAGGAG CAAGGTGAGA TGACAGGAGA TCCTGCCCCG
  841 GCACTTCGCC CAATAGCAGC CAGTCCCTTC CCGCTTCAGT GACAACGTCG AGCACAGCTG
  901 CGCAAGGAAC GCCCGTCGTG GCCAGCCACG ATAGCCGCGC TGCCCTCGTC TTGCAGTTCA
  961 TTCAGGGCAC CCGGACAGGT CGGTCTTGAC AAAAAGAACC CGGGCGCCCC CTGCGCTGAC
 1021 AGCCCGGGAA CGCGCGGCAT CAGAAGCAGC CCGATTTGTC TGTTGTGCCC AGTCATAAGC
1081 CGATAGCCTC TTCCACCCAG CGGCCGGGAG AAAACCCTTG GCGGTTGCAA CATTC
//

Please visit Sequencing and PCR primers for primer information.


References

References

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.

2018.09.03

GNP_filter3_RDBDEP_html_180823.pl