RDB14423 - pT7-hCas9-mC-polyA

Please review the QC test results indicated by check

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

pT7-hCas9-mC-polyA (#RDB14423)

Cas9-poly(A) expressing improved plasmid.

Alternative name	T7-NLS-hCAS9-Cdt1-NLS-pA
Clone info.	This version contains no FLAG-tag. If you want to have a FLAG-tag version, please order RDB14604.
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 Mouse Cdt1 cDNA
Depositor\|Developer	DNA Bank, \|

Sequence (full)	RDB14423zzt02.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.

提供案内 (日本国内) [open/close]

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.)
RDB14423	pT7-hCas9-mC-polyA	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 pT7-hCas9-mC-polyA was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB14423).

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	RDB14423_A6L1p1-1.pdf

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

Primer: CMV_Forward (Pr0016) Region: CMV pro, T7 pro, SV40 NLS, insert5'(Cas9 5') Sequence file: RDB14423_A6L1a.seq
>14423_14423_B2FgF_1_CMV_Forward_F03_18_ABI24.ab1 1 NNNNNNNNNN NNNNNNTTGN NGCNNNTGGG CGGTAGGCGT GTACGGTGGG AGGTCTATAT 61 AAGCAGAGCT CGTTTAGTGA ACCGTCAGAT CGCCTGGAGA CGCCATCCAC GCTGTTTTGA 121 CCTCCATAGA AGACACCGGG ACCGATCCAG CCTCCGGACT CTAGTAATAC GACTCACTAT 181 AGGGAGAGCC GCCACCATGG CCCCAAAGAA GAAGCGGAAG GTCGGTATCC ACGGAGTCCC 241 AGCAGCCGAC AAGAAGTACT CCATTGGGCT CGATATCGGC ACAAACAGCG TCGGCTGGGC 301 CGTCATTACG GACGAGTACA AGGTGCCGAG CAAAAAATTC AAAGTTCTGG GCAATACCGA 361 TCGCCACAGC ATAAAGAAGA ACCTCATTGG CGCCCTCCTG TTCGACTCCG GGGAGACGGC 421 CGAAGCCACG CGGCTCAAAA GAACAGCACG GCGCAGATAT ACCCGCAGAA AGAATCGGAT 481 CTGCTACCTG CAGGAGATCT TTAGTAATGA GATGGCTAAG GTGGATGACT CTTTCTTCCA 541 TAGGCTGGAG GAGTCCTTTT TGGTGGAGGA GGATAAAAAG CACGAGCGCC ACCCAATCTT 601 TGGCAATATC GTGGACGAGG TGGCGTACCA TGAAAAGTAC CCAACCATAT ATCATCTGAG 661 GAAGAAGCTT GTAGACAGTA CTGATAAGGC TGACTTGCGG TTGATCTATC TCGCGCTGGC 721 GCATATGATC AAATTTCGGG GANACTTCCT CATCGAGGGG GANCTGAACC CAGACAACAG 781 CGATGTCGAC AAACTCTTTA TCCAACTGGT TCAGANTTAC AATCAGCTTT TCGAAGAGAA 841 NCCGATCNAC GCATCCGGAG TTGANGCCNA AGCAATCCTG ANCGCTANGN TGTNNNANTC 901 CNGGCGGCTC GAAAANCNCA TCGCACAGCN CCCNGGGGAN AANAANAACG GNCNGNTTGG 961 TANTCNNANC GCCCTGNCNC TCGGGNNGAN CCCCNACTTT AAATCTNANT TCGNNCNNNN 1021 NNANNNNNNN NNN //
Primer: HSV-pA_R (Pr0949) Region: SV40 NLS, insert3' (ccdt1, Cas9 3') Sequence file: RDB14423_A6L1b.seq
>14423_14423_B2FgF_1_HSV-pA_R_G03_21_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNN GNGTTNNNGT TAGCCTCCCC CGTTNAAACT CNTTACTAAC 61 CGGTAGGGAT CGAACCCTTT CACACCTTCC TCTTCTTCTT GGGGTCAGCC CTGCTGACGG 121 AGTCCTCTGA GGGGATCTTG CCCGGCTGTT GACCAACATA AACAGTTGTA CTCTTTGTTC 181 GCTTGACGGG GGATGGGCAA ACGGGGGAGC CAGGGTCAGC TGGAGGGCTA GGGTCAGCTG 241 GAGGGCTGGG CTCAGCTGGG GAACTGGGCT CAGGCAGAGA ACTGGGGCAG GAGTCCAATC 301 CAGGCAGCCG TAACCTCCGG CGCGCGAGCG GCGCGGGCTG GTCACTCCCG GGTTCGGCTG 361 GGGGCCGGGC GCGCTTGCGG CTGCTGCTCC GGGTGAACGC AGGAGCCACG AGGCCACCAG 421 GGCTCGGGTC TCCACCGAGC TGAGAGAGGT CGATTCTTGT TTCATAGAGC CCCGTAATTG 481 ACTGATGAAT CAGTGTGGCG TCCAGGACCT CCTTTGTAGA GGTGTACCGC TTTCTGTCTA 541 TGGTGGTGTC GAAGTACTTG AAGGCTGCAG GCGCGCCCAA GTTGGTCAGA GTAAACAAGT 601 GGATAATGTT TTCTGCCTGC TCCCTGATGG GCTTATCCCT GTGCTTATTG TAAGCAGAAA 661 GCACCTTATC GAGGTTAGCG TCGGCGAGGA TCACTCTTTT GGAGAATTCG CTTATTTGCT 721 CGATGATCTC ATCAAGGTAG TGTTTGTGTT GTTCCACGAA CAGCTGCTTC TGCTCATTAT 781 CTTCGGGAGA CCCTTTGAGC TTTTCATAGT GGCTGGNCAG ATACAAGAAA TTAACGTATT 841 TAGAGGGCAG TGCCAGCTCG TTACNTTTCT GCAGCTCGCC CGCACTAGCG AGCATTCGTT 901 TCCGGNCGTT TTCAAGCTCN AAGANAGAGN ANTTNGGAAG CTNNNTGATG AGGNCTTTTT 961 TTGANNNCTT TNTATCCNTT CNNNNCNAGA AANTCNNNGG GGNNTTTTTT TCNAANNNNG 1021 NTNNNNNNNN NANNGNGANN NCNNNNNNNN NN //
Primer: f1ori_F (Pr0280) Region: f1 ori, pA Sequence file: RDB14423_A6L1c.seq
>14423_14423_B2FgF_1_f1ori_F_H03_24_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNGN ANNGNAGCGG TCACGCTGCG CGTAACCACC ACACCCGCCG 61 CGCTTAATGC GCCGCNACNG GGCGCGTGGG GATACCCCCT AGAGCCCCAG CTGCGCAGAT 121 CTGCTATGGC AGGGCCTGCC GCCCCGACGT TGGCTGCGAG CCCTGGGCCT TCACCCGAAC 181 TTGGGGGGTG GGGTGGGGAA AAGGAAGAAA CGCGGGCGTA TTGGCCCCAA TGGGGTCTCG 241 GTGGGGTATC GATGTACAGC TAGCTTTTTT TTTTTTTTTT TTTTTTTTTT TTTTTTTTTT 301 TTTTTTTTTT TTTTTTTTTT TTTTTTTTTT TTTTTTTTTN NNNNNNNNN //
Primer: SV40pro_F (Pr0110) Region: SV40pro_ori, NeoR Sequence file: RDB14423_A6L1d.seq
>D04203A5_A6L1_1_SV40pro_F_B06_06_ABI24.ab1 1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNTG NNGANGCTTT TTTGGAGGCC TAGGCTTTTG 61 CAAAAAGCTC CCGGGAGCTT GTATATCCAT TTTCGGATCT GATCAAGAGA CAGGATGAGG 121 ATCGTTTCGC ATGATTGAAC AAGATGGATT GCACGCAGGT TCTCCGGCCG CTTGGGTGGA 181 GAGGCTATTC GGCTATGACT GGGCACAACA GACAATCGGC TGCTCTGATG CCGCCGTGTT 241 CCGGCTGTCA GCGCAGGGGC GCCCGGTTCT TTTTGTCAAG ACCGACCTGT CCGGTGCCCT 301 GAATGAACTG CAGGACGAGG CAGCGCGGCT ATCGTGGCTG GCCACGACGG GCGTTCCTTG 361 CGCAGCTGTG CTCGACGTTG TCACTGAAGC GGGAAGGGAC TGGCTGCTAT TGGGCGAAGT 421 GCCGGGGCAG GATCTCCTGT CATCTCACCT TGCTCCTGCC GAGAAAGTAT CCATCATGGC 481 TGATGCAATG CGGCGGCTGC ATACGCTTGA TCCGGCTACC TGCCCATTCG ACCACCAAGC 541 GAAACATCGC ATCGAGCGAG CACGTACTCG GATGGAAGCC GGTCTTGTCG ATCAGGATGA 601 TCTGGACGAA GAGCATCAGG GGCTCGCGCC AGCCGAACTG TTCGCCAGGC TCAAGGCGCG 661 CATGCCCGAC GGCGAGGATC TCGTCGTGAC CCATGGCGAT GCCTGCTTGC CGAATATCAT 721 GGTGGAAAAT GGNCGCTTTT CTGGATTCAT CGACTGTGGC CGGCTGGGTG TGGCGGANCG 781 CTATCAGGAC ATAGCGTTGG CTACCCGTGA TATTGCTGAA GAGCTTGGCG GCGAATGGGC 841 TGANNGCTTC CTCGTGCTTT ACGGTATCGC CGCTCCCGAT TCGCAGCGCA TCGCCTTCTA 901 TCGCCTNNTT GACGAGTTCT TCTGANCGGG ANNCTNGGGN NCNCGAANTG ANNNANCNAG 961 CGANGCCNNA NCNNGCCNTC ACNANATTTC NANNNCNNNN NNNNCNTNNN ATGNAANNNN 1021 NGGNNNNNNN NNN //

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]