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

pMK312 (#RDB18362)

Expression vector of Cas9 and gRNA for a TOP2A tagging donor.

Alternative name TOP2A-CRISPR
Clone info. Expression vector of Cas9 and gRNA for a TOP2A tagging donor, see Fig. 5 and SFig. 8 of Yesbolatova, A. et al., Nat. Commun. 11: 5701, 2020.
Vector backbone px330 (plasmid)
Selectable markers Ampicillin
Gene/insert name human TOP2A cDNA
Depositor|Developer Kanemaki, Masato |
 
Sequence (full) RDB18362zzk01.seq provided by the depositor

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
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Exclusive MTA (For the DNA materials containing CRISPR/Cas9 technologies and for not-for-profit academic purpose) [Word]
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 literature designated by the DEPOSITOR is requested (Yesbolatova, A. et al., Nat. Commun. 11: 5701 (2020)).
Remarks
The BIOLOGICAL RESOURCE contains CRISPR/Cas9 technologies and is not provided to for-profit organization or for for-profit research by non-profit organizations.
提供案内 (日本国内) [open/close]

必要書類
提供依頼書  [依頼書の記入例 ]
専用MTA（CRISPR/Cas9内包遺伝子材料専用 非営利学術目的）をお使いください [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 (Yesbolatova, A. et al., Nat. Commun. 11: 5701 (2020))。
備考
本件リソースはCRISPR/Cas9 technologyを用いたゲノム編集バイオリソースです。営利機関および非営利機関による営利目的研究には提供いたしません。

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB18362 pMK312 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 pMK312 was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB18362).

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 RDB18362_B1K1p1-1.pdf check

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

Primer: U6pro_F3 (Pr0648)
Region: U6pro, TOP2A taget seq, gRNA scaffold, CMV en, chicken beta-actin pro
Sequence file: RDB18362_B1K1a.seq check
>18362_18362_B1K1_2_U6pro_F3_A02_02_ABI24.ab1
    1 NNNNNNNNNN NNNNGGNNNA TNNTATGCTT ACCGTAACTT GAAAGTATTT CGATTTCTTG
   61 GCTTTATATA TCTTGTGGAA AGGACGAAAC ACCGGACTCT TCCAGGTACT TTATGTTTTA
  121 GAGCTAGAAA TAGCAAGTTA AAATAAGGCT AGTCCGTTAT CAACTTGAAA AAGTGGCACC
  181 GAGTCGGTGC TTTTTTGTTT TAGAGCTAGA AATAGCAAGT TAAAATAAGG CTAGTCCGTT
  241 TTTAGCGCGT GCGCCAATTC TGCAGACAAA TGGCTCTAGA GGTACCCGTT ACATAACTTA
  301 CGGTAAATGG CCCGCCTGGC TGACCGCCCA ACGACCCCCG CCCATTGACG TCAATAGTAA
  361 CGCCAATAGG GACTTTCCAT TGACGTCAAT GGGTGGAGTA TTTACGGTAA ACTGCCCACT
  421 TGGCAGTACA TCAAGTGTAT CATATGCCAA GTACGCCCCC TATTGACGTC AATGACGGTA
  481 AATGGCCCGC CTGGCATTGT GCCCAGTACA TGACCTTATG GGACTTTCCT ACTTGGCAGT
  541 ACATCTACGT ATTAGTCATC GCTATTACCA TGGTCGAGGT GAGCCCCACG TTCTGCTTCA
  601 CTCTCCCCAT CTCCCCCCCC TCCCNACCCC CNATTTTGTA TTTATTTATT TTTTAATTAN
661 TTTGTGCAGC GANGGGNNNN NNNNNN
//
Primer: Cas9-N_R1 (Pr0650)
Region: CAS9 5', SV40 NSL, 3xFLAG, hybrid intron
Sequence file: RDB18362_B1K1b.seq check
>18362_18362_B1K1_2_Cas9-N_R1_B02_05_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNTGNAC TCGTCGGTGA NCNCGGCCCN GCCCACAGAG
   61 TTGGTGCCGA TGTCCAGGCC GATGCTGTAC TTCTTGTCGG CTGCTGGGAC TCCGTGGATA
  121 CCGACCTTCC GCTTCTTCTT TGGGGCCATC TTATCGTCAT CGTCTTTGTA ATCAATATCA
  181 TGATCCTTGT AGTCTCCGTC GTGGTCCTTA TAGTCCATGG TGGCACCGGT CCAACCTGAA
  241 AAAAAGTGAT TTCAGGCAGG TGCTCCAGGT AATTAAACAT TAATACCCCA CCAACCAACC
  301 ATCCCTTAAA CCCTTACCTC TTGCTCAGCT AATTACAGCC CGGAGGAGAA GGGCCGTCCC
361 GCCCGCTCAC CTGTGGGAGT AACGCGGTCA GTCAGAGCCG GGGCGGGCGG CGCGAGGN
//
Primer: Cas9-C_F2 (Pr1051)
Region: CAS9 3', nucleoplasmin NLS, bGH pA, AAV2 ITR
Sequence file: RDB18362_B1K1c.seq check
>18362_18362_B1K1_2_Cas9-C_F2_C02_08_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNC CCTNNTCNNC CNGAGCATCN CCGGCCTGTA CGAGACACGG
   61 ATCGACCTGT CTCAGCTGGG AGGCGACAAA AGGCCGGCGG CCACGAAAAA GGCCGGCCAG
  121 GCAAAAAAGA AAAAGTAAGA ATTCCTAGAG CTCGCTGATC AGCCTCGACT GTGCCTTCTA
  181 GTTGCCAGCC ATCTGTTGTT TGCCCCTCCC CCGTGCCTTC CTTGACCCTG GAAGGTGCCA
  241 CTCCCACTGT CCTTTCCTAA TAAAATGAGG AAATTGCATC GCATTGTCTG AGTAGGTGTC
  301 ATTCTATTCT GGGGGGTGGG GTGGGGCAGG ACAGCAAGGG GGAGGATTGG GAAGAGAATA
  361 GCAGGCATGC TGGGGAGCGG CCGCAGGAAC CCCTAGTGAT GGAGTTGGCC ACTCCCTCTC
421 TGCGCGCN
//
Primer: f1ori_F (Pr0280)
Region: f1 ori, AAV2 ITR
Sequence file: RDB18362_B1K1d.seq check
>18362_18362_B1K1_2_f1ori_F_D02_11_ABI24.ab1
    1 NNNNNNNNNN NNGCNNGNAN NGNAGCGGTN ACGCTGCGCG TAACCACCAC ACCCGCCGCG
   61 CTTAATGCGC CGCTACAGGG CGCGTACTAT GGTTGCTTTG ACGTATGCGG TGTGAAATAC
  121 CGCACAGATG CGTAAGGAGA AAATACCGCA TCAGGCGCCC CTGCAGGCAG CTGCGCGCTC
181 NNTCGCTCAC TGNNGCNNNC CNGNNNNNNN NNN
//

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]