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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

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

Catalog number RDB18362
Resource name pMK312
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 |
Other clones in our bank
External Database
human TOP2A
          Reference sequence
            
           
          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.
          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)).
          Ordering Please visit Information of Request for Distribution.[link] 
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          Exclusive MTA (For the DNA materials containing CRISPR/Cas9 technologies and for not-for-profit academic purpose) [Word]
          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]

          提供条件 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))。
          提供依頼 手続きの詳細は、「提供申込みについて[link]」をご覧ください。
          提供依頼書 [Word]
          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]
          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

          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.

          Further references such as user reports and related articles (go to bottom)

          References and tips

          Featured content

          Featured content Auxin Inducible Degron (AID) System (English text)
          Featured content Auxin Inducible Degron (AID) System (Japanese text)

          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.

          2023.05.01

          GNP_filter3_RDBDEP_html_230501.pl