Resource data sheet
Prev.

T7-NLS hCas9-pA

Cas9-poly(A) expressing improved plasmid.

Catalog number RDB13130
Resource name T7-NLS hCas9-pA
Clone info. Cas9 expressing plasmid. Induction with CMV or T7 promoter. Applicable for both direct transfection into mammalian cells and T7 in-vitro translation.
Digest with Nhe I for linearization prior to in vitro transcription followed by capping and polyadenylation.
Vector backbone pcDNA3 (Plasmid)
Selectable markers Amp^r
Gene/insert name Streptococcus pyogenes Cas9 genomic DNA
Depositor Mashimo, Tomoji |
 
Other clones in our bank Streptococcus pyogenes (KEGG orthology K09952) |

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 Information [in Japanese] [in English]
Terms and conditions set forth by the DEPOSITOR In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature designated by the DEPOSITOR is requested (Yoshimi, K. et al. Nat. Commun. 7: 10431, 2016). RECIPIENT which wants to use the BIOLOGICAL RESOURCE for the purpose other than education or not-for-profit research is requested to enter into a Material Transfer Agreement with Osaka University.
(Japanese text [open/close])
Remarks
Please use the Exclusive MTA (genome editing) [Word] [pdf]
(Japanese text [open/close])

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB13130 T7-NLS hCas9-pA DNA solution

Sequence information

RIKEN BRC has sequenced portions of this material for quality test. Primers and the results are shown below.

Data are summerized on test sheet below.

Test sheet RDB13130_A5Bkp1.pdf

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

Primer: CMV-Forward RDB13130_A5Bka.seq
>13130_13130_A5Bk_CMV_Foward_3_H02_23.ab1
    1 TTGTACACTC CGCCCCATTG ACGCAAATGG 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 GGACCTGAAC CCAGACAACA
  781 GCGATGTCGA CAAACTCTTT ATCCAACTGG TTCAGACTTA CAATCAGCTT TTCGAAGAGA
  841 ACCCGATCAA CGCATCCGGA GTTGACGCCA AAGCAATCCT GAGCGCTAGG CTGTCCAAAT
  901 CCCGGCGGCT CGAAAACCTC ATCGCACAGC TCCCTGGGGA GAAGAAGAAC GGCCTGTTTG
  961 GTAATCTTAT CGCCCTGTCA CTCGGGCTGA CCCCCAACTT TAAATCTAAC TTCGACCTGG
 1021 TCGAGATGCC AAGCTTTCAC TGAGCAAGAC ACCTACGATG ATGATCTCGA CATCTGCTGC
 1081 CAGATCGGCG ACCAGTTACG CAGACGTTTT TTTTGGCCGC AAGACTGTCA GAACGCCCAA
1141 TTCCTTGACT
//
Primer: HSV TK pA_R RDB13130_A5Bkb.seq
>13130_13130_A5Bk_HSV_tk_pA_R_3_A03_03.ab1
    1 ATGGAATCTT CGGTATTGTC TCCTTCCGTG TTTCAGTTAG CCTCCCCCGT TTAAACTCAT
   61 TACTAACCGG TAGGGATCGA ACCCTTTCAC ACCTTCCTCT TCTTCTTGGG GTCAGCCCTG
  121 CTGTCTCCAC CGAGCTGAGA GAGGTCGATT CTTGTTTCAT AGAGCCCCGT AATTGACTGA
  181 TGAATCAGTG TGGCGTCCAG GACCTCCTTT GTAGAGGTGT ACCGCTTTCT GTCTATGGTG
  241 GTGTCGAAGT ACTTGAAGGC TGCAGGCGCG CCCAAGTTGG TCAGAGTAAA CAAGTGGATA
  301 ATGTTTTCTG CCTGCTCCCT GATGGGCTTA TCCCTGTGCT TATTGTAAGC AGAAAGCACC
  361 TTATCGAGGT TAGCGTCGGC GAGGATCACT CTTTTGGAGA ATTCGCTTAT TTGCTCGATG
  421 ATCTCATCAA GGTAGTGTTT GTGTTGTTCC ACGAACAGCT GCTTCTGCTC ATTATCTTCG
  481 GGAGACCCTT TGAGCTTTTC ATAGTGGCTG GCCAGATACA AGAAATTAAC GTATTTAGAG
  541 GGCAGTGCCA GCTCGTTACC TTTCTGCAGC TCGCCCGCAC TAGCGAGCAT TCGTTTCCGG
  601 CCGTTTTCAA GCTCAAAGAG AGAGTACTTG GGAAGCTTAA TGATGAGGTC TTTTTTGACC
  661 TCTTTATATC CTTTCGCCTC GAGAAAGTCG ATGGGGTTTT TTTCGAAGCT TGATCGCTCC
  721 ATGATTGTGA TGCCCAGCAG TTCCTTGACG CTTTTGAGTT TTTTAGACTT CCCTTTCTCC
  781 ACTTTGGCCA CAACCAGTAC ACTGTAAGCG ACTGTAGGAG AATCGAATCC GCCGTATTTC
  841 TTGGGGTCCC AATCTTTTTT GCGTGCGATC AGCTTGTCGC TGTTCCTTTT CGGGAGGATA
  901 CTTTCCTTGG AGAAGCCTCC GGTCTGTACT TCGGTCTTTT TAACGATGTT CACCTGCGGC
  961 ATGGACAGGA CCTTCCGGGA CTGTCGCGAA ATCCCTACCC TTTGTCCCAC ACGATTTCTC
 1021 CTGTTTCTCC GTTGTTTTCG ATAAGTGGTC GCCTTCGATC TCTCATTGGT CAGTGTATCT
 1081 CGGTCTTCGA AAAACTCATA TATGCTGTAA GAGGTACCTA GCGATGGCTT GCTAATTTCT
1141 GCCTCAGGAA CTTTTGGGCC GGAT
//
Primer: SV40pro_F RDB13130_A5Bkc.seq
>13130_13130_A5Bk_SV40pro_F_3_B03_06.ab1
    1 CCCTGTCCTG GGCATTCAGA GTAGTGAGGA GGCTTTTTTG GAGGCCTAGG CTTTTGCAAA
   61 AAGCTCCCGG GAGCTTGTAT ATCCATTTTC GGATCTGATC AAGAGACAGG ATGAGGATCG
  121 TTTCGCATGA TTGAACAAGA TGGATTGCAC GCAGGTTCTC CGGCCGCTTG GGTGGAGAGG
  181 CTATTCGGCT ATGACTGGGC ACAACAGACA ATCGGCTGCT CTGATGCCGC CGTGTTCCGG
  241 CTGTCAGCGC AGGGGCGCCC GGTTCTTTTT GTCAAGACCG ACCTGTCCGG TGCCCTGAAT
  301 GAACTGCAGG ACGAGGCAGC GCGGCTATCG TGGCTGGCCA CGACGGGCGT TCCTTGCGCA
  361 GCTGTGCTCG ACGTTGTCAC TGAAGCGGGA AGGGACTGGC TGCTATTGGG CGAAGTGCCG
  421 GGGCAGGATC TCCTGTCATC TCACCTTGCT CCTGCCGAGA AAGTATCCAT CATGGCTGAT
  481 GCAATGCGGC GGCTGCATAC GCTTGATCCG GCTACCTGCC CATTCGACCA CCAAGCGAAA
  541 CATCGCATCG AGCGAGCACG TACTCGGATG GAAGCCGGTC TTGTCGATCA GGATGATCTG
  601 GACGAAGAGC ATCAGGGGCT CGCGCCAGCC GAACTGTTCG CCAGGCTCAA GGCGCGCATG
  661 CCCGACGGCG AGGATCTCGT CGTGACCCAT GGCGATGCCT GCTTGCCGAA TATCATGGTG
  721 GAAAATGGCC GCTTTTCTGG ATTCATCGAC TGTGGCCGGC TGGGTGTGGC GGACCGCTAT
  781 CAGGACATAG CGTTGGCTAC CCGTGATATT GCTGAAGAGC TTGGCGGCGA ATGGGCTGAC
  841 CGCTTCCTCG TGCTTTACGG TATCGCCGCT CCCGATTCGC AGCGCATCGC CTTCTATCGC
  901 CTTCTTGACG AGTTCTTCTG AGCGGGACTC TGGGGTTCGC GAAATGACCG ACCAAGCGAC
  961 GCCCAACCCT GCCATCACGA GATTTCGATT CCACCGCCGC TTCTATGAAA GGTTGGCTTC
 1021 GGATCGTTTT CCGGGACGCC GGCTGAATGA TCTCAGCGCG GGGATCTCAT GCCTGAAGTC
 1081 CTCGGCCAAC CCCACTGGTA TGCAGCTATA TGGTTACTAT AAGGCCATTA GCCATCCACC
1141 AACTTCCCCA AACTTAAAG
//

Please visit plasmidSequencing and PCR primers for primer information.

References and tips

Electronic file

Featured content

Featured content Genome Editing (English text)
Featured content Genome Editing (Japanese text)

References

original Yoshimi, K., ssODN-mediated knock-in with CRISPR-Cas for large genomic regions in zygotes. Nat. Commun. 7: 10431 (2016). PMID 26786405.

2017.10.19

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