RDB14602 - T7-3xFLAG-NLS-hCAS9-NLS-pA

Please review the QC test results indicated by check

icon below as well as clone information before placing your order.

T7-3xFLAG-NLS-hCAS9-NLS-pA (#RDB14602)

Cas9-poly(A) expressing improved plasmid.

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, \|

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.

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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 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.

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.

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Ordering forms	Order form [Credit Card Payment] [Bank Transfer Payment] [Example of order form ] 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	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.
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.

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	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。
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	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 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. [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	RDB14602_A7B9p1-1.pdf

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
>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
>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
>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
>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
>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
>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. [link to RRC of NBRP]