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T7-3xFLAG-NLS-hCAS9-NLS-pA

Cas9-poly(A) expressing improved plasmid.

Catalog number RDB14602
Resource name T7-3xFLAG-NLS-hCAS9-NLS-pA
Clone info. FLAG-tagged Cas9 expressing plasmid. Induction with CMV or T7 promoter. Applicable for both direct transfection into mammalian cells and T7 in-vitro translation.
Comment Constructed from T7-NLS hCas9-pA (RDB13130, Yoshimi, K., Nat. Commun. 7: 10431, 2016. PMID 26786405).
Vector backbone pcDNA3 (Plasmid)
Selectable markers Amp^r
Gene/insert name Streptococcus pyogenes Cas9 genomic DNA
Depositor|Developer DNA Bank, |
Other clones in our bank

External Database
Streptococcus pyogenes Cas9

          Reference sequence
            
           
          Sequence (full) RDB14602zzt02.seq compiled by us

          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 The use of the BIOLOGICAL RESOURCE is restricted to the academic researches conducted by non-profit organization. In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR (Mizuno-Iijima, S., et al., Methods 191: 23-31 (2021)) is requested. The RECIPIENT agrees to expressly describe the acknowledgment of the RIKEN BRC as the source of the BIOLOGICAL RESOURCE in any publication.
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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 本件リソースの使用は学術機関での学術研究に限る。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 (Mizuno-Iijima, S., et al., Methods 191: 23-31 (2021)) を引用する。本件リソースを使用して研究成果を発表する場合、理研BRCから提供されたことを明示する。
          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.)
          RDB14602 T7-3xFLAG-NLS-hCAS9-NLS-pA 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 T7-3xFLAG-NLS-hCAS9-NLS-pA was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB14602).

          Reference section:

          Mizuno-Iijima, S., Ayabe, S., Kato, K., Matoba, S., Ikeda, Y., Dinh, T.T.H., Le, H.T., Suzuki, H., Nakashima, K., Hasegawa, Y., Hamada, Y., Tanimoto, Y., Daitoku, Y., Iki, N., Ishida, M., Ibrahim, E.A.E., Nakashiba., , T., Hamada, M., Murata, K., Miwa, Y., Okada-Iwabu, M., Iwabu, M., Yagami, K., Ogura, A., Obata, Y., Takahashi, S., Mizuno, S., Yoshiki, A., Sugiyama, F., Efficient production of large deletion and gene fragment knock-in mice mediated by genome editing with Cas9-mouse Cdt1 in mouse zygotes. Methods 191: 23-31 (2021). PMID 32334080.

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


          References and tips

          Featured content

          Featured content Genome Editing (English text)
          Featured content Genome Editing (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 RDB14602_A7B9p1-1.pdf check

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

          Primer: CMV_Forward (Pr0016)
          Region: CMV pro,T7 pro,3XFLAG,SV40 NLS,insert 5'
          Sequence file: RDB14602_A7B9a.seq check
          >D04438A1_A7B9_1_CMV-Forward_F08_17_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN NGNNGCNNAT GGGCGGTAGG CGTGTACGGT GGGAGGTCTA
             61 TATAAGCAGA GCTCGTTTAG TGAACCGTCA GATCGCCTGG AGACGCCATC CACGCTGTTT
            121 TGACCTCCAT AGAAGACACC GGGACCGATC CAGCCTCCGG ACTCTAGTAA TACGACTCAC
            181 TATAGGGAGA GCCGCCACCA TGGACTATAA GGACCACGAC GGAGACTACA AGGATCATGA
            241 TATTGATTAC AAAGACGATG ACGATAAGGC CCCAAAGAAG AAGCGGAAGG TCGGTATCCA
            301 CGGAGTCCCA GCAGCCGACA AGAAGTACTC CATTGGGCTC GATATCGGCA CAAACAGCGT
            361 CGGCTGGGCC GTCATTACGG ACGAGTACAA GGTGCCGAGC AAAAAATTCA AAGTTCTGGG
            421 CAATACCGAT CGCCACAGCA TAAAGAAGAA CCTCATTGGC GCCCTCCTGT TCGACTCCGG
            481 GGAGACGGCC GAAGCCACGC GGCTCAAAAG AACAGCACGG CGCAGATATA CCCGCAGAAA
            541 GAATCGGATC TGCTACCTGC AGGAGATCTT TAGTAATGAG ATGGCTAAGG TGGATGACTC
            601 TTTCTTCCAT AGGCTGGAGG AGTCCTTTTT GGTGGAGGAG GATAAAAAGC ACGAGCGCCA
            661 CCCAATCTTT GGCAATATCG TGGACGANGT GGCGTACCAT GAAAAGTACC CNACCATATN
            721 TNNTCTGANG AAGAANCTTG TANNCNGTAC TGATAAGGCT GACTTGCGGT TGATCTATCT
            781 CGCGCTGGCG CATATGATCA AATTTCGGGG ANNCTTCCNC ATCGAGGGGG ANCTGAANCC
            841 NGACANCAGC GATGNCGACA AACTCTTTAT CCNNCNGGTT NNNNNNTTNC NANCAGCNTT
            901 TNCGANNANA NNCNGATCNA CGCNTNCNGN ANTTGNNGCC NANNNNNTCC TGANCGCTNN
          961 NNNNTNNNAA NNCCNNNGNN NNNNNNNN
          //
          Primer: HSV_TK_pA_R (Pr0137)
          Region: SV40 NLS,insert 3'
          Sequence file: RDB14602_A7B9b.seq check
          >D04438A1_A7B9_1_HSV_TK_pA_R_G08_20_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNNTCC TTCCGTGTTN NNNTTAGCCT CCNNNNNNTA
             61 AACTCATTAC TAACCGGTAG GGATCGAACC CTTTCACACC TTCCTCTTCT TCTTGGGGTC
            121 AGCCCTGCTG TCTCCACCGA GCTGAGAGAG GTCGATTCTT GTTTCATAGA GCCCCGTAAT
            181 TGACTGATGA ATCAGTGTGG CGTCCAGGAC CTCCTTTGTA GAGGTGTACC GCTTTCTGTC
            241 TATGGTGGTG TCGAAGTACT TGAAGGCTGC AGGCGCGCCC AAGTTGGTCA GAGTAAACAA
            301 GTGGATAATG TTTTCTGCCT GCTCCCTGAT GGGCTTATCC CTGTGCTTAT TGTAAGCAGA
            361 AAGCACCTTA TCGAGGTTAG CGTCGGCGAG GATCACTCTT TTGGAGAATT CGCTTATTTG
            421 CTCGATGATC TCATCAAGGT AGTGTTTGTG TTGTTCCACG AACAGCTGCT TCTGCTCATT
            481 ATCTTCGGGA GACCCTTTGA GCTTTTCNNA NTGGCTGGCC AGATNNNNNA AATTAACGTA
            541 TTTAGAGGGC AGTGCCAGCT CGTTACCTTT CTGCAGCTCG CCCGCACTAG CNNNNNNTCG
            601 TTTCCGGCCG TTTTCAAGCT CAAAGAGAGA GTACTNGNNA NGCTTANNGA TGAGGTCTTT
            661 TTTGACCTCT TTATATCCTT TCNNCTCNAN NNNGTCGATG GGGTTTTTTT CGAAGCTTGA
            721 TCNNNNNNNT GATTGTGATG NCCAGCNNTT CCTTGACGCT TTTGAGTTTT TTAGACTTCC
            781 CTTTCTCCAC TTTGGCCACA ACCAGTACAC TGTAAAGCNN ANTGTANGAN AATCGANTCC
            841 GCCGNATTNN TTGGGGGNNN CCNNNNCNTT TTTTGCGNGC NATCAGNTTN GTCGNNGNTN
            901 NNNTTTTCNG NAGGANNNTT TCNNTNNGNA NNNNCCTCCG GGTCNGTACT TCNGNNNTTT
          961 TTNNNGANNN TNNNCNTNGN NGGNNNNNGN NNAGGNACNN NNNNNNGNAN NNNNNNNNN
          //
          Primer: hCAS9+2010/F (Pr0425)
          Region: insert mid
          Sequence file: RDB14602_A7B9c.seq check
          >D04438A1_A7B9_1_hCAS9+2010_F_H08_23_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNNNNNGAT TTGNNNNNCG GAACTTCATG
             61 CAGTTGATCC ATGATGACTC TCTCACCTTT AAGGAGGACA TCCAGAAAGC ACAAGTTTCT
            121 GGCCAGGGGG ACAGTCTTCA CGAGCACATC GCTAATCTTG CAGGTAGCCC AGCTATCAAA
            181 AAGGGAATAC TGCAGACCGT TAAGGTCGTG GATGAACTCG TCAAAGTAAT GGGAAGGCAT
            241 AAGCCCGAGA ATATCGTTAT CGAGATGGCC CGAGAGAACC AAACTACCCA GAAGGGACAG
            301 AAGAACAGTA GGGAAAGGAT GAAGAGGATT GAAGAGGGTA TAAAAGAACT GGGGTCCCAA
            361 ATCCTTAAGG AACACCCAGT TGAAAACACC CAGCTTCAGA ATGAGAAGCT CTACCTGTAC
            421 TACCTGCAGA ACGGCAGGGA CATGTACGTG GATCAGGAAC TGGACATCAA TCGGCTCTCC
            481 GACTACGACG TGGATCATAT CGTGCCCCAG TCTTTTCTCA AAGATGATTC TATTGATAAT
            541 AAAGTGTTGA CAAGATCCGA TAAAAATAGA GGGAAGAGTG ATAACGTCCC CTCAGAANAA
            601 NTTGTCAAGA AAATGAAAAA TTATTGGCGG CAGCTGCTGA ACGCCAAACT GATCACACAA
            661 CGGAAGTTCG ATAATCTGAC TAAGGCTGAA CGAGGTGGCC TGTCTGAGTT GGATAAAGCC
            721 GGCTTCNTCA AAAGGCAGCT TGTTGAGACA CGCCAGATCA CCAAGCACGT GGCCCAAATT
            781 CTCGATTCAC GCNTGAACAC CAAGNACGAT GAAAATGACA AACTGATTCG ANAGGTGAAA
            841 GTTATTACTC TGAAGTCTAA GCTGGTCTCN NANNTCNGAA AANGANTTTC NNTTTTATNA
            901 GGNGNNANAN ATNANCNNTN ANCNNNANNG CGCANGATGC CTACCTGGAN TGNAGNGGNN
            961 NNGCNCTGNN NTNNTCAAAA AATATNCCNN NNNNTNGNAN NNGNNNTTTG NTTNNCNGNN
           1021 NCTATNNNNN GNNCNNANNN TNGNNAAANN NATNNNNNAA GNNNNNNNNN GNNNNNNNNN
          1081 N
          //
          Primer: hCAS9+3550/R (Pr0424)
          Region: insert mid
          Sequence file: RDB14602_A7B9d.seq check
          >D04438A1_A7B9_1_hCAS9+3550_R_A09_03_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNGA NNGNGNNGCC CNNNNNNTCC TTGNCGCTTT
             61 TGAGTTTTTT AGACTTCCCT TTCTCCACTT TGGCCACAAC CAGTACACTG TAAGCGACTG
            121 TAGGAGAATC GAATCCGCCG TATTTCTTGG GGTCCCAATC TTTTTTGCGT GCGATCAGCT
            181 TGTCGCTGTT CCTTTTCGGG AGGATACTTT CCTTGGAGAA GCCTCCGGTC TGTACTTCGG
            241 TCTTTTTAAC GATGTTCACC TGCGGCATGG ACAGGACCTT CCGGACTGTC GCGAAATCCC
            301 TACCCTTGTC CCACACGATT TCTCCTGTTT CTCCGTTTGT TTCGATAAGT GGTCGCTTCC
            361 GAATCTCTCC ATTGGCCAGT GTAATCTCGG TCTTGAAAAA ATTCATAATA TTGCTGTAAA
            421 AGAAGTACTT AGCGGTGGCC TTGCCTATTT CCTGCTCAGA CTTTGCGATC ATTTTCCTAA
            481 CATCGTACAC TTTATAGTCT CCGTAAACAA ATTCAGATTC AAGCTTGGGA TATTTTTTGA
            541 TAAGTGCAGT GCCTACCACT GCATTCAGGT AGGCATCATG CGCATGGTGG TAATTGTTGA
            601 TCTCTCTCAC CTTATAAAAC TGAAAGTCCT TTCTGAAATC TGAGACCAGC TTAGACTTCA
            661 GAGTAATAAC TTTCACCTCT CGAATCAGTT TGTCATTTTC ATCGTACTTG GTGTTCATGC
            721 GTGAATCGAG AATTTGGGCC ACGTGCTTGG TGATCTGGCG TGTCTCAACA AGCTGCCTTT
            781 TGANGANNNN GGCTTTATCN NACTCANACA NGGCCACCTC GTTCANCCTT ANNCAGANTA
            841 TCGAACTTCC GTTGTGTGAT CAGTTNNGNN GTTCANCANC TGNCGCNNAT ANTTTTTCAT
            901 TTTNCTNGAC ANNTTCTTNT GANGGGANGT TNNNACNCTN NCCTCTATTT TNATCGGANN
            961 TTGNCNNNCT TTNNNATCNN NANANNCATC TTNNNNNNAN NNNNGGGGGN NNNANNTGAT
          1021 CCNNNNNNNA NNNNNANANN CNANNNNNNG NNNNNNNNNN
          //
          Primer: f1ori_F (Pr0280)
          Region: f1 ori,p A
          Sequence file: RDB14602_A7B9e.seq check
          >D04438A1_A7B9_1_f1ori_F_B09_06_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNGCGNNNNN NCTGCGCGTA ACCACCACAC
             61 CCGCCGCGCT TAATGCGCCG CTACAGGGCG CGTGGGGATA CCCCCTAGAG CCCCAGCTGC
            121 GCAGATCTGC TATGGCAGGG CCTGCCGCCC CGACGTTGGC TGCGAGCCCT GGGCCTTCAC
            181 CCGAACTTGG GGGGTGGGGT GGGGAAAAGG AAGAAACGCG GGCGTATTGG CCCCAATGGG
            241 GTCTCGGTGG GGTATCGATG TACAGCTAGC TTTTTTTTTT TTTTTTTTTT TTTTTTTTTT
          301 TTTTTTTTTT TTTTTTTTTT TTTTTTTTTT TTTTTTTTTT TTTTTTTTNN NNNNNNNN
          //
          Primer: SV40pro_F (Pr0110)
          Region: NeoR
          Sequence file: RDB14602_A7B9f.seq check
          >D04438A1_A7B9_1_SV40pro_F_C09_09_ABI24.ab1
              1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNGNGGANGN TTTTTTGGAG GNCTAGGCTT
             61 TTGCAAAAAG CTCCCGGGAG CTTGTATATC CATTTTCGGA TCTGATCANN ANANNGGATG
            121 AGGATCGTTT CGCATGATTG AACAAGATGG ATTGCACGCA GGTTCTCCGG CCGCTTGGGT
            181 GGAGAGGCTA TTCGGCTATG ACTGGGCACA ACAGACAATC GGCTGCTCTG ATGCCGCCGT
            241 GTTCCGGCTG TCAGCGCAGG GGCGCCCGGT TCTTTTTGTC AAGACCGACC TGTCCGGTGC
            301 CCTGAATGAA CTGCAGGACG AGGCAGCGCG GCTATCGTGG CTGGCCACGA CGGGCGTTCC
            361 TTGCGCAGCT GTGCTCGACG TTGTCACTGA AGCGGGAAGG GACTGGCTGC TATTGGGCGA
            421 AGTGCCGGGG CAGGATCTCC TGTCATCTCA CCTTGCTCCT GCCGAGAAAG TATCCATCAT
            481 GGCTGATGCA ATGCGGCGGC TGCATACGCT TGATCCGGCT ACCTGCCCAT TCGACCACCA
            541 AGCGAAACAT CGCATCGAGC GAGCACGTAC TCGGATGGAA GCCGGTCTTG TCGATCAGGA
            601 TGATCTGGAC GAAGAGCATC AGGGGCTCGC GCCAGCCGAA CTGTTCGCCA GGCTCAAGGC
            661 GCGCATGCCC GACGGCGAGG ATCTCGTCGT GACCCATGGC GATGCCTGCT TGCCGAATAT
            721 CATGGTGGAA AATGNCCGCT TTTCTGGATT CATCGACTGT GGCCGGCTGG GTGTGGCGGA
            781 NCGCTATCAG GACATAGCGT TGGCTACNCG TGATATTGCT GAAGANCTTG NCGGCGAATG
            841 GGCTGACCGC TTCCTCGTGC TTTACGGTAT CGCCGCTCCC GATTCGCANC GCATCGCCTT
            901 CTATCGCCNT NCTTGACNAN NNCTTCTGAN CGNNACTCTG GGGTTCGCGA AATGANCGAN
          961 NNAGCGACNN CCNANCNNNN NNTCNCNANN NTTCNATTNN NNNNNNNN
          //

          Please visit Sequencing and PCR primers for primer information.


          References

          Original, user report and related articles

          original Mizuno-Iijima, S., Efficient production of large deletion and gene fragment knock-in mice mediated by genome editing with Cas9-mouse Cdt1 in mouse zygotes. Methods 191: 23-31 (2021). PMID 32334080.

          2023.05.01

          GNP_filter3_RDBDEP_html_230501.pl