Resource data sheet
DNA Bank Top
Please review the QC test results indicated by check icon below as well as clone information before placing your order.

T7-NLS-hCAS9(ver1.1)-NLS-pA (#RDB14420)

Cas9-poly(A) with enhanced specificity.

Clone info. Cas9 expressing plasmid. Induction with CMV or T7 promoter. Applicable for both direct transfection into mammalian cells and T7 in-vitro translation. Having enhanced specificity version (SpCas9-K848A,K1003A,R1060A; Slaymaker IM et al., science 351(6268), 84-88, 2016: PMID26628643).
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) RDB14420zzt02.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.
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. The RECIPIENT agrees to expressly describe the acknowledgement 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]

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。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.)
RDB14420 T7-NLS-hCAS9(ver1.1)-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-NLS-hCAS9(ver1.1)-NLS-pA was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB14420).

Reference section:

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 RDB14420_A6L1p1-1.pdf check

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

Primer: CMV_Forward (Pr0016)
Region: CMV pro, SV40 NLS, insert 5'
Sequence file: RDB14420_A6L1a.seq check
>D04203A2_A6L1_1_CMV_Forward_C06_03_ABI08.ab1
    1 NNNNNNNNNN NNNCNNNNTT GNNGCNNNGG GCGGTAGGCG TGTACGGTGG GAGGTCTATA
   61 TAAGCAGAGC TCGTTTAGTG AACCGTCAGA TCGCCTGGAG ACGCCATCCA CGCTGTTTTG
  121 ACCTCCATAG AAGACACCGG GACCGATCCA GCCTCCGGAC TCTAGTAATA CGACTCACTA
  181 TAGGGAGAGC CGCCACCATG GCCCCAAAGA AGAAGCGGAA GGTCGGTATC CACGGAGTCC
  241 CAGCAGCCGA CAAGAAGTAC TCCATTGGGC TCGATATCGG CACAAACAGC GTCGGCTGGG
  301 CCGTCATTAC GGACGAGTAC AAGGTGCCGA GCAAAAAATT CAAAGTTCTG GGCAATACCG
  361 ATCGCCACAG CATAAAGAAG AACCTCATTG GCGCCCTCCT GTTCGACTCC GGGGAGACGG
  421 CCGAAGCCAC GCGGCTCAAA AGAACAGCAC GGCGCAGATA TACCCGCAGA AAGAATCGGA
  481 TCTGCTACCT GCAGGAGATC TTTAGTAATG AGATGGCTAA GGTGGATGAC TCTTTCTTCC
  541 ATAGGCTGGA GGAGTCCTTT TTGGTGGAGG AGGATAAAAA GCACGAGCGC CACCCAATCT
  601 TTGGCAATAT CGTGGACGAG GTGGCGTACC ATGAAAAGTA CCCAACCATA TATCATCTGA
  661 GGAAGAAGCT TGTAGACAGT ACTGATAAGG CTGACTTGCG GTTGATCTAT CTCGCGCTGG
  721 CGCATATGAT CAAATTTCGG GGACACTTCC TCATCGAGGG GGANCTGAAC CCAGACAACA
  781 GCGNTGTCGA CNAACTCTTT ATCCNACTGG NTCNNANTTA CAATCAGCTT TTCGAAGANN
  841 AACCCGATCA ACGCATCCNG AGTTGANGCC NAAGCNATCC TGANCGCTNN GCTGNNNNAA
  901 TCCCNGNCGG NTCNAAAANC TCNTCGCACA NCTCCCNGGG GNNNNNAAGN NNNGNCNNNT
961 NNNNNNNNNN
//
Primer: HSV_TK_pA_R (Pr0137)
Region: SV40 NLS, insert 3'
Sequence file: RDB14420_A6L1b.seq check
>D04203A2-1_A6L1_1_HSV_TK_pA_R_A02_02_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNTG NCNNNTTCCG TGTTTNNGTT AGCCTCCCCC GTTTAAACTC
   61 ATTACTAACC GGTAGGGATC GAACCCTTTC ACACCTTCCT CTTCTTCTTG GGGTCAGCCC
  121 TGCTGTCTCC ACCGAGCTGA GAGAGGTCGA TTCTTGTTTC ATAGAGCCCC GTAATTGACT
  181 GATGAATCAG TGTGGCGTCC AGGACCTCCT TTGTAGAGGT GTACCGCTTT CTGTCTATGG
  241 TGGTGTCGAA GTACTTGAAG GCTGCAGGCG CGCCCAAGTT GGTCAGAGTA AACAAGTGGA
  301 TAATGTTTTC TGCCTGCTCC CTGATGGGCT TATCCCTGTG CTTATTGTAA GCAGAAAGCA
  361 CCTTATCGAG GTTAGCGTCG GCGAGGATCA CTCTTTTGGA GAATTCGCTT ATTTGCTCGA
  421 TGATCTCATC AAGGTAGTGT TTGTGTTGTT CCACGAACAG CTGCTTCTGC TCATTATCTT
  481 CGGGAGACCC TTTGAGCTTT TCATAGTGGC TGGCCAGATA CAAGAAATTA ACGTATTTAG
  541 AGGGCAGTGC CAGCTCGTTA CCTTTCTGCA GCTCGCCCGC ACTAGCGAGC ATTCGTTTCC
  601 GGCCGTTTTC AAGCTCAAAG AGAGAGTACT TGGGAAGCTT AATGATGAGG TCTTTTTTGA
  661 CCTCTTTATA TCCTTTCGCC TCGAGAAAGT CGATGGGGTT TTTTTCGAAG CTTGATCGCT
  721 CCATGATTGT GATGCCCAGC AGTTCCTTGA CGCTNNTGAN TTTTTTAGAC TTCCCTTTCT
  781 CCACTTNGGC CACNACCAGT ACACTGNAAG CGACTGTANG AGAATCNANT CCGCCGTATT
  841 TCTTGGGGNC CNAATCTTTT TTNGCGTGCN ATCAGCTNGN CGCNNTNCCT TTNNGGGANG
  901 ANNNTTTCNT NNNNNAANNN NNNNGNCTGN ACNTCGGNNN TTTTANNGAN GNTNNNNNNN
961 NNN
//
Primer: hCAS9+2010_F (Pr0425)
Region: insert mid (K848A)
Sequence file: RDB14420_A6L1c.seq check
>D04203A2_A6L1_1_hCAS9+2010_F_D06_04_ABI08.ab1
    1 NNNNNNNNNN NNNNNNTNNN NNNNNNCCGA NGGNNTTGCC AACCGGAACT TCATGCAGTT
   61 GATCCATGAT GACTCTCTCA CCTTTAAGGA GGACATCCAG AAAGCACAAG TTTCTGGCCA
  121 GGGGGACAGT CTTCACGAGC ACATCGCTAA TCTTGCAGGT AGCCCAGCTA TCAAAAAGGG
  181 AATACTGCAG ACCGTTAAGG TCGTGGATGA ACTCGTCAAA GTAATGGGAA GGCATAAGCC
  241 CGAGAATATC GTTATCGAGA TGGCCCGAGA GAACCAAACT ACCCAGAAGG GACAGAAGAA
  301 CAGTAGGGAA AGGATGAAGA GGATTGAAGA GGGTATAAAA GAACTGGGGT CCCAAATCCT
  361 TAAGGAACAC CCAGTTGAAA ACACCCAGCT TCAGAATGAG AAGCTCTACC TGTACTACCT
  421 GCAGAACGGC AGGGACATGT ACGTGGATCA GGAACTGGAC ATCAATCGGC TCTCCGACTA
  481 CGACGTGGAT CATATCGTGC CCCAGTCTTT TCTCGCAGAT GATTCTATTG ATAATAAAGT
  541 GTTGACAAGA TCCGATAAAA ATAGAGGGAA GAGTGATAAC GTCCCCTCAG AAGAAGTTGT
  601 CAAGAAAATG AAAAATTATT GGCGGCAGCT GCTGAACGCC AAACTGATCA CACAACGGAA
  661 GTTCGATAAT CTGACTAAGG CTGAACGAGG TGGCCTGTCT GANTTGGATA AAGCCGGCTT
  721 CATCAAAAGG CAGCTTGTTG AGACACGCCA GATCACCAAG CACGTGGCCC AAATTCTCGA
  781 NTCACGCATG AACACCANGT ACGATGANAA TGACNAACTG ATTCGAGAGG TGAAAGTTAT
  841 TACTCTGAAG TCTAAGCTGG TCTCAGATTT CAGAAAGGAC TTTCNGTTTT ATANNGNGAG
  901 AGAGATCAAC NNTTACNNCN NNGCGCATGA NGNCNACCNN NANGCNNNGG TANGNNNNGN
961 NCNTNNTCAA AAAATATCCN NNGNNNNNNN NNN
//
Primer: hCAS9+3550_R (Pr0424)
Region: insert mid (R1060A, K1003A)
Sequence file: RDB14420_A6L1d.seq check
>D04203A2_A6L1_1_hCAS9+3550_R_E06_05_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNGNNNG NGNNGCCCAG CANNTTCCTT GNNGCTTTTG
   61 AGTTTTTTAG ACTTCCCTTT CTCCNCTTTG GCCACAACCA GTACACTGTA AGCGACTGTA
  121 GGAGAATCGA ATCCGCCGTA TTTCTTGGGG TCCCAATCTT TTTTGCGTGC GATCAGCTTG
  181 TCGCTGTTCC TTTTCGGGAG GATACTTTCC TTGGAGAAGC CTCCGGTCTG TACTTCGGTC
  241 TTTTTAACGA TGTTCACCTG CGGCATGGAC AGGACCTTCC GGACTGTCGC GAAATCCCTA
  301 CCCTTGTCCC ACACGATTTC TCCTGTTTCT CCGTTTGTTT CGATAAGTGG TGCCTTCCGA
  361 ATCTCTCCAT TGGCCAGTGT AATCTCGGTC TTGAAAAAAT TCATAATATT GCTGTAAAAG
  421 AAGTACTTAG CGGTGGCCTT GCCTATTTCC TGCTCAGACT TTGCGATCAT TTTCCTAACA
  481 TCGTACACTT TATAGTCTCC GTAAACAAAT TCAGATTCAA GCGCGGGATA TTTTTTGATA
  541 AGTGCAGTGC CTACCACTGC ATTCAGGTAG GCATCATGCG CATGGTGGTA ATTGTTGATC
  601 TCTCTCACCT TATAAAACTG AAAGTCCTTT CTGAAATCTG AGACCAGCTT ANACTTCAGA
  661 GTANTAACTT TCACCTCTCG AATCAGTTTG TCATTTTCAT CGTACTTGGT GTTCATGCGT
  721 GAATCGAGAA TTTGGGCCAC GTGCTTGGTG ATCTGGCGTG TCTCANCNAG CTGCCTTTTG
  781 ATGAANCCGG CTTTATCNNA CTCANANAGG CCNCCTCGTT CAGCCTTANT CAGANTATCG
  841 AACTTCCGTT GTGTGATCAG TTTGNNNNNN CANCAGCTGN CGCNNANTAA TTTTTTNNNT
  901 TTNCNTGANA NNTNCNTCNG AGGGGNANGT TNNNNNCTCN NCCNTCNNNT TTTNNNCNGN
961 ATNNNNGNNN NNNNNTTNNN NATNNNNANN ANNNNNNNNN N
//
Primer: SV40pro_ori_F (Pr0731)
Region: NeoR
Sequence file: RDB14420_A6L1e.seq check
>D04203A2_A6L1_1_SV40pro_ori_F_F06_06_ABI08.ab1
    1 NNNNNNNNNN NNNNNNNNNN GCCTCTGCCT CTGNNCTANT CNNNNNNNAG TGAGGAGGCT
   61 TTTTTGGAGG CCTAGGCTTT TGCANANNNC TCCCGGGAGC TTGTATATCC ATTTTCGGAT
  121 CTGATCAAGA GACAGGATGA GGATCGTTTC GCATGATTGA ACAAGATGGA TTGCACGCAG
  181 GTTCTCCGGC CGCTTGGGTG GAGAGGCTAT TCGGCTATGA CTGGGCACAA CAGACAATCG
  241 GCTGCTCTGA TGCCGCCGTG TTCCGGCTGT CAGCGCAGGG GCGCCCGGTT CTTTTTGTCA
  301 AGACCGACCT GTCCGGTGCC CTGAATGAAC TGCAGGACGA GGCAGCGCGG CTATCGTGGC
  361 TGGCCACGAC GGGCGTTCCT TGCGCAGCTG TGCTCGACGT TGTCACTGAA GCGGGAAGGG
  421 ACTGGCTGCT ATTGGGCGAA GTGCCGGGGC AGGATCTCCT GTCATCTCAC CTTGCTCCTG
  481 CCGAGAAAGT ATCCATCATG GCTGATGCAA TGCGGCGGCT GCATACGCTT GATCCGGCTA
  541 CCTGCCCATT CGACCACCAA GCGAAACATC GCATCGAGCG AGCACGTACT CGGATGGAAG
  601 CCGGTCTTGT CGATCAGGAT GATCTGGACG AAGAGCATCA NGGGCTCGCG CCAGCCGAAC
  661 TGTTCGCCAG GCTCAAGGCG CGCATGCCCG ACGGCGAGGA TCTCGTCGTG ACCCATGGCG
  721 ATGCCTGCTT GCCGAATATC ATGGTGGAAA ATGGCCGCTT TTCTGGATTC NTCGACTGTG
  781 GCCGGCTGGG NGTGGNGGAN CGCTATCANG ACATANCGTT GGCTACCCGN GATATTGCTG
  841 AAGANCTNGN CGGCGAANGG GCTGANNNCT TCCTCGTGCT TTNNCNGNAT CNNCNGCTCC
  901 CGATTCNNAN CGCATCNNNT TCTATCNNNN TCTNGACNAN TNCNTCNNNN CGGNNNTNNG
961 GGGGNNNNNN NNAANNNNNN NNNN
//

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles