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

pMK322 (#RDB18361)

Donor plasmid to generate TOP2A-mAID degron cell lines with mClover as a marker, hygromycin resistance.

Alternative name TOP2A-mAC(Hygro)
Clone info. Donor plasmid to generate TOP2A-mAID degron cell lines with mClover as a marker, hygromycin resistance, see Fig. 5 and SFig. 8 of Yesbolatova, A. et al., Nat. Commun. 11: 5701, 2020.
Vector backbone pBluescriptII KS- (plasmid)
Selectable markers Ampicillin (E. coli), Hygromycin (mammalian cell).
Gene/insert name human TOP2A genomic Arabidopsis thaliana IAA17 cDNA Aequorea victoria GFP (mClover) cDNA
Depositor|Developer Kanemaki, Masato |
 
Sequence (full) RDB18361zzk01.seq provided by the depositor

Distribution information

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MTA, for use for not-for-profit academic purpose [Word] [Example of MTA ]
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Terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested (Yesbolatova, A. et al., Nat. Commun. 11: 5701 (2020)).
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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 ]
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 (Yesbolatova, A. et al., Nat. Commun. 11: 5701 (2020))。

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB18361 pMK322 DNA solution JPY 9,460 (not-for-profit academic 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 pMK322 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB18361).

Reference section:

Yesbolatova, A., Saito, Y., Kitamoto, N., Makino-Itou, H., Ajima, R., Nakano, R., Nakaoka, H., Fukui, K., Gamo, K., Tominari, Y., Takeuchi, H., Saga, Y., Hayashi, K., Kanemaki, M.T., The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. Nat. Commun. 11: 5701 (2020). PMID 33177522. [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 RDB18361_B1K1p1-1.pdf check

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

Primer: pUC-CAPbs_up_F2 (Pr0739)
Region: hTOP2A_1, linker, mAID 5'
Sequence file: RDB18361_B1K1a.seq check
>18361_18361_B1K1_3_pUC-CAPbs_up_F2_A01_01_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNTNNNNNNN TTATGCTTCC GGCTCGTATG TTGTGTGGAA
   61 TTGTGAGCGG ATAACAATTT CACACAGGAA ACAGCTATGA CCATGATTAC GCCAAGCTCG
  121 GCGCGCCATT GGGATGGAAC GAAAACTCCT TGAATATAGT GTCTTGATAC ATTTTTTTAC
  181 ATTACAATTA TGTTGTCAGA TTTACAATGT GCAAGTTACC TGGGCTTTTC TCTTTTAGAA
  241 ATCCAAGGGG GAGAGTGATG ACTTCCATAT GGACTTTGAC TCAGCTGTGG CTCCTCGGGC
  301 AAAATCTGTA CGGGCAAAGA AACCCATTAA ATACCTGGAA GAGTCAGATG AAGATGATCT
  361 GTTTGGATCC GGTGCAGGCG CCAAGGAGAA GAGTGCTTGT CCTAAAGATC CAGCCAAACC
  421 TCCGGCCAAG GCACAAGTTG TGGGATGGCC ACCGGTGAGA TCATACCGGA AGAACGTGAT
  481 GGTTTCCTGC CAAAAATCAA GCGGTGGCCC GGAGGCGGCG GCGTTCGTGA AGGTATCAAT
  541 GGACGGAGCA CCGTACTTGA NGAAAATCGA TTTGAGGATG TATAAAGCTA GCATGGTGAG
  601 CAAGGGCGAN GANCTGTTCN CCGGGGTGGT GNCCATCCNG NNCGAGCTGG ANGGCGACNT
  661 AAACNGNCNC NAGTTCNNCG TNCGCNGCNA GGGNNANGGC NANGCCNCNA NNNNNNNGCT
721 NANCCTGAAN TTNNNCNNNN NNNNNN
//
Primer: pUC-M13_-100 (Pr0310)
Region: hTOP2A_2, loxP-2, mPGK ter
Sequence file: RDB18361_B1K1b.seq check
>18361_18361_B1K1_3_pUC-M13_-100_B01_04_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNTTTCCNN GTCNCGACGT TGTAAAACGA CGGCCAGTGA
   61 ATTGACGCGT ATTGGGATGG AACGCCCATG AGATGGTCAC TATTTAGACA GTATTATAAA
  121 AAGCTAAAGA ACACTTGGGC TTTACTTCAC TTTGATGTCT TGTACTAAAA ACACCTTCCC
  181 CAAACTAAAT TCAGAGGGGA GGAAGTTAAG AGCTTCAGGT AACTTTAAAA CCAGTCTTGG
  241 GCTTGGTAAG ATAATTACTT AAAATAATCG CCTCACATTT TAAAACAGAT CATCTTCATC
  301 TGACTCTGGA TCCTAGTGAA CCTCTTCGAG GGACCTAATA ACTTCGTATA GCATACATTA
  361 TACGAAGTTA TATTAAGGGT TCCGGATCTC GACCAGCTTC TGATGGAATT AGAACTTGGC
  421 AAAACAATAC TGAGAATGAA GTGTATGTGG AACAGAGGCT GCTGATCTCG TTCTTCAGGC
  481 TATGAAACTG ACACATTTGG AAACCNNNNN NCTTAGAACC ACAAAGTGGG AATCAAGAGA
  541 AAAACAATGA TCCCACGAGA GATCTATAGA TCTATAGATC ATGAGTGGGA GGAANGAGCT
  601 GGNCCTTAAT TNGGNTTNGN NTGNTTAANT NANNATATCN NANTATGAAA NNNTATCNTA
  661 AAGNANNNNN NNAAGAGCCT TCNNTAAAGA NNNNNNNTTN NTCTNATCCN NNCCCNNNCC
721 CCNNNCCCNN NNNNNNNCNN NNNNCTTNNN NNNNNNN
//
Primer: PGKpro_R (Pr0793)
Region: PGK pro, loxP-1. SV40 pA, mClover(A206K) 3'
Sequence file: RDB18361_B1K1c.seq check
>18361_18361_B1K1_3_PGKpro_R_C11_08_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNG NNGNGTGCGA NGCCNGANGC CNNTTGTGTA GCGCCNAGTG
   61 CCCAGCGGGG CTGCTAAAGC GCATGCTCCA GACTGCCTTG GGAAAAGCGC CTCCCCTACC
  121 CGGTAGAATT TCGAGGTCGA GATCCTAAGC TTGGCTGGAC GTAAACTCCT CTTCAGACCT
  181 AATAACTTCG TATAGCATAC ATTATACGAA GTTATATTAA GGGTTATTGA ATATGATCGA
  241 CGCGTTAAGA TACATTGATG AGTTTGGACA AACCACAACT AGAATGCAGT GAAAAAAATG
  301 CTTTATTTGT GAAATTTGTG ATGCTATTGC TTTATTTGTA ACCATTATAA GCTGCAATAA
  361 ACAAGTTAAC AACAACAATT GCATTCATTT TATGTTTCAG GTTCAGGGGG AGGTGTGGGA
  421 GGTTTTTTAA AGCAAGTAAA ACCTCTACAA ATGTGGTATG GCTGATTATG ATCAGTTATC
  481 TAGATTACTT GTACAGCTCG TCCATGCCAT GTGTAATCCC GGCGGCGGTC ACGAACTCCA
  541 GCAGGACCAT GTGATCGCGC TTCTCGTTGG GGTCTTTGCT CAGCTTGGAC TGATGGCTCA
  601 GGTAGTGGTT GTCGGGCAGC AGCACGGGGC CGTCGCCGAT GGGGGTGTTC TGCTGGTAGT
  661 GGTCGGCGAG CTGCACGCTG CCGTCCTCAA CGTTGTGGCG GATCTTGAAG TTAGCCTTGA
  721 TGCCGTTCTT CTGCTTGTCG GCCGTGATAT AGACGTTGTG GCTGTTGAAG TTGTACTCCA
  781 GCTTGTGCCN CANGATGTTG CCGTCCTCCT TGAAGTCGAT GCCCTTCAGC TCGATGCGGT
  841 TCACCNNGGT GTCGCNCTCN AACTTCANCT CNGCGCGGGT CTTGNNNNNA CCNNCGNNNN
  901 NGAAAGAGAN GGTGCGNTNN TGGACGTANC CNTCNGGCNT GGNNGANTTN NNNNNGNNGN
961 GCTNCNNCAT GNGNNNNGGG NANCNGNNTG NNNNNNNNNN
//
Primer: PGKpro_F (Pr0249)
Region: HygR
Sequence file: RDB18361_B1K1d.seq check
>18361_18361_B1K1_3_PGKpro_F_E01_13_ABI24.ab1
    1 NNNNNNNNNN NNNNNNTCTC CNGGCCTTTC GACCTGCATC CNTCTAGATC TCGAGCAGCT
   61 GAAGCTTACC GCTAGCATGG ATAGATCCGG AAAGCCTGAA CTCACCGCGA CGTCTGTCGA
  121 GAAGTTTCTG ATCGAAAAGT TCGACAGCGT CTCCGACCTG ATGCAGCTCT CGGAGGGCGA
  181 AGAATCTCGT GCTTTCAGCT TCGATGTAGG AGGGCGTGGA TATGTCCTGC GGGTAAATAG
  241 CTGCGCCGAT GGTTTCTACA AAGATCGTTA TGTTTATCGG CACTTTGCAT CGGCCGCGCT
  301 CCCGATTCCG GAAGTGCTTG ACATTGGGGA ATTCAGCGAG AGCCTGACCT ATTGCATCTC
  361 CCGCCGTGCA CAGGGTGTCA CGTTGCAAGA CCTGCCTGAA ACCGAACTGC CCGCTGTTCT
  421 GCAGCCGGTC GCGGAGGCCA TGGATGCGAT CGCTGCGGCC GATCTTAGCC AGACGAGCGG
  481 GTTCGGCCCA TTCGGACCGC AAGGAATCGG TCAATACACT ACATGGCGTG ATTTCATATG
  541 CGCGATTGCT GATCCCCATG TGTATCACTG GCAAACTGTG NTGGACGACA CCGTCAGTGC
  601 GTCCGTCGCG CANGCTCTCG ATGANCTGAN GCTTTGGGCC GANGANTGNC CCNAAGTCCG
  661 GCACCTCNNG CACNCNGNNT TCNGCTNCNN CNNTGNCCNG ANNNNNNNNN GNNNGCNTNN
721 NNNNNGTCNN NNNNNNNN
//
Primer: EGFP-C-rev (Pr0026)
Region: mClover(A206K) 5', mAID 3'
Sequence file: RDB18361_B1K1e.seq check
>18361_18361_B1K1_3_EGFP-C-rev_F01_06_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNTTGGNN TGNNGGCTCN NNGNNNNGGT
   61 TGTCGGGCAG CANCNCGGGG CCGTCGCCGA TGGGGGTGTT CTGCTGGTAG TGGTCGGCGA
  121 GCTGCACGCT GCCGTCCTCA ACGTTGTGGC GGATCTTGAA GTTAGCCTTG ATGCCGTTCT
  181 TCTGCTTGTC GGCCGTGATA TAGACGTTGT GGCTGTTGAA GTTGTACTCC AGCTTGTGCC
  241 CCAGGATGTT GCCGTCCTCC TTGAAGTCGA TGCCCTTCAG CTCGATGCGG TTCACCAGGG
  301 TGTCGCCCTC GAACTTCACC TCGGCGCGGG TCTTGTAGGT ACCGTCGTCC TTGAAAGAGA
  361 TGGTGCGCTC CTGGACGTAG CCTTCGGGCA TGGCGGACTT GAAGAAGTCG TGCTGCTTCA
  421 TGTGGTCGGG GTAGCGGCTG AAGCAGGCCA CGCCGTAGCC GAAGGTGGTC ACGAGGGTGG
  481 GCCAGGGCAC GGGCAGCTTG CCGGTGGTGC AGATGAACTT CAGGGTCAGC TTGCCGTTGG
  541 TGGCATCGCC CTCGCCCTCG CCGCGGACGC TGAACTTGTG GCCGTTTACG TCGCCGTCCA
  601 GCTCGACCAG GATGGGCACC ACCCCGGTGA ACAGCTCCTC GCCCTTGCTC ACCATGCTAG
  661 CTTTATACAT CCTCAAATCG ATTTTCCTCN AGTACGGTGC TCCGTNNNNN GATACCTTCA
  721 CGAACGCCGC CGCCTCCGGG CCACCGCTTG ATTTTTGGCA NGAAANCATC ACGTTCTTCC
  781 GGTATGATCT CNNCGGNGGC CATCNCNCNN CTTGTGCCTT GGCCGGANGT TTGGCTGGAT
  841 CTTTANGANA AGCACTCNTT CTCCTNNGCG NCNGCACCGG NNCCAAACAG NATCATCTTC
  901 ATCTGACTNT NNNANGTATT TNANNGGGGN TTCTTTNNCC CGNNNNNGAT TTTNNNNNNN
  961 AGGANNNNNC NNGNNGNANN NNNNANNNCN ANATNGNAAA NNCNNTNNNC NNNNNNNNNN
//

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original Yesbolatova, A., The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. Nat. Commun. 11: 5701 (2020). PMID 33177522. [link to RRC of NBRP]