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pCS2P-krtt1c19e-linker-eGFP-mut-donor (#RDB13481)

Donor vector for integrating gRNA-resistant eGFP mutant into zebrafish krtt1c19e locus

Clone info. Donor vector for integrating gRNA-resistant eGFP mutant into zebrafish krtt1c19e locus
Comment CMV promoter of the pCS2p vector was removed by HindIII-SalI digestion.
Vector backbone pCS2p (plasmid)
Size of vector backbone 3.0 kb
Selectable markers Amp^r
Gene/insert name Zebrafish krtt1c19e genomic DNA Aquorea victoria Green flurescent protein (GFP) cDNA
Depositor|Developer Kawahara, Atsuo | Hisano, Yu |
 
Sequence RDB13481z.seq

Distribution information

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MTA, for use for not-for-profit academic purpose [Word] [Example of MTA ]
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Terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature(s) designated by the DEPOSITOR is requested. (Sci. Rep., 5, 8841 (2015))
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提供依頼書  [依頼書の記入例 ]
提供同意書 (MTA, 非営利学術目的用)[Word] [MTAの記入例 ]
手続きの概要は、「提供申込みについて[link]」をご覧ください。営利目的利用についてはお問い合わせください。
MTAに書く使用条件 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を引用する。(Sci Rep., 5, 8841 (2015))

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB13481 pCS2P-krtt1c19e-linker-eGFP-mut-donor 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 pCS2P-krtt1c19e-linker-eGFP-mut-donor was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB13481).

Reference section:

Hisano, Y., Sakuma, T., Nakade, S., Ohga, R., Ota, S., Okamoto, H., Yamamoto, T., Kawahara, A., Precise in-frame integration of exogenous DNA mediated by CRISPR/Cas9 system in zebrafish. Sci. Rep. 5: 8841 (2015). PMID 25740433. [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 RDB13481_A5L8p1-3.pdf check

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

Primer: f1ori_R3 (Pr0142)
Region: eGFP-gRNA,homology arm,linker,eGFP,SV40p(A)
Sequence file: RDB13481_A5L8c.seq check
>13481_13481_A5L8_f1ori_R3_G12_21.ab1
    1 GGAATGCCCA TTCAGGCTGC GCACTGTTGG GAGGGCGATC GGTGCGGGCC TCTTCGCTAT
   61 TACGCCAGTC GAGGCGAGGG CGATGCCACC TACGGAGAGG TGATTGATGG AAAGGTGGTT
  121 TCTTCCTCTT CCACGGGAGG AGGTGGTTCA GGTGGTGGAG GATCTGGAGG TGGAGGTTCA
  181 GAATTCATGG TGAGCAAGGG CGAGGAGCTG TTCACCGGGG TGGTGCCCAT CCTGGTCGAG
  241 CTGGACGGCG ACGTAAACGG CCACAAGTTC AGCGTGTCCG GCGAGGGCGA AGGGGACGCT
  301 ACATACGGCA AGCTGACCCT GAAGTTCATC TGCACCACCG GCAAGCTGCC CGTGCCCTGG
  361 CCCACCCTCG TGACCACCCT GACCTACGGC GTGCAGTGCT TCAGCCGCTA CCCCGACCAC
  421 ATGAAGCAGC ACGACTTCTT CAAGTCCGCC ATGCCCGAAG GCTACGTCCA GGAGCGCACC
  481 ATCTTCTTCA AGGACGACGG CAACTACAAG ACCCGCGCCG AGGTGAAGTT CGAGGGCGAC
  541 ACCCTGGTGA ACCGCATCGA GCTGAAGGGC ATCGACTTCA AGGAGGACGG CAACATCCTG
  601 GGGCACAAGC TGGAGTACAA CTACAACAGC CACAACGTCT ATATCATGGC CGACAAGCAG
  661 AAGAACGGCA TCAAGGTGAA CTTCAAGATC CGCCACAACA TCGAGGACGG CAGCGTGCAG
  721 CTCGCCGACC ACTACCAGCA GAACACCCCC ATCGGCGACG GCCCCGTGCT GCTGCCCGAC
  781 AACCACTACC TGAGCACCCA GTCCGCCCTG AGCAAAGACC CCAACGAGAA GCGCGATCAC
  841 ATGGTCCTGC TGGAGTTCGT GACCGCCGCC GGGATCACTC TCGGCATGGA CGAGCTGTAC
  901 AAGTAATCTA GATTCTGCAG CCCTATAGTG AGTCGTATTA CGTAGATCCA GACATGATAA
  961 GATACATTGA TGAGTTTGGG ACAAACCACA ACTAGAATGC AGTGAAAAAA ATGCTTTATT
 1021 TGTGAAATTT GTGATGCTAT TGCTTTATTT GTAACCAATA ATAAGCTGCA TAACAAGCTA
1081 ACCAACAACA ACTGGCCAAT TTCAATTTTT ATGGATTTCC AGCTTTCCAG
//
Primer: Reverse-2 (Pr0100)
Region: eGFP-gRNA,homology arm,SV40p(A),eGFP
Sequence file: RDB13481_A5L8d.seq check
>13481_13481_A5L8_Reverse2_H12_24.ab1
    1 ATTTCCATGA ATTACGCCAA GCGCGCAATT AACCCTCACT GGCGAGGGCG ATGCCACCTA
   61 CGGTTTACCC GACATGAATC TGGTGTGCTT TACTTAACAA GGGCACACCT CCCCCTGAAC
  121 CTGAAACATA AAATGAATGC AATTGTTGTT GTTAACTTGT TTATTGCAGC TTATAATGGT
  181 TACAAATAAA GCAATAGCAT CACAAATTTC ACAAATAAAG CATTTTTTTC ACTGCATTCT
  241 AGTTGTGGTT TGTCCAAACT CATCAATGTA TCTTATCATG TCTGGATCTA CGTAATACGA
  301 CTCACTATAG GGCTGCAGAA TCTAGATTAC TTGTACAGCT CGTCCATGCC GAGAGTGATC
  361 CCGGCGGCGG TCACGAACTC CAGCAGGACC ATGTGATCGC GCTTCTCGTT GGGGTCTTTG
  421 CTCAGGGCGG ACTGGGTGCT CAGGTAGTGG TTGTCGGGCA GCAGCACGGG GCCGTCGCCG
  481 ATGGGGGTGT TCTGCTGGTA GTGGTCGGCG AGCTGCACGC TGCCGTCCTC GATGTTGTGG
  541 CGGATCTTGA AGTTCACCTT GATGCCGTTC TTCTGCTTGT CGGCCATGAT ATAGACGTTG
  601 TGGCTGTTGT AGTTGTACTC CAGCTTGTGC CCCAGGATGT TGCCGTCCTC CTTGAAGTCG
  661 ATGCCCTTCA GCTCGATGCG GTTCACCAGG GTGTCGCCCT CGAACTTCAC CTCGGCGCGG
  721 GTCTTGTAGT TGCCGTCGTC CTTGAAGAAG ATGGTGCGCT CCTGGACGTA GCCTTCGGGC
  781 ATGGCGGACT TGAAGAAGTC GTGCTGCTTC ATGTGGTCGG GGTAGCGGCT GAAGCACTGC
  841 ACGCCGTAGT CAGGGGTGGT CACGAGGGTG GGCCAGGGCA CGGGCAGCTT GCCGGTGGTG
  901 CAGATGAACT TCAGGGTCAG CTTGCCGTAT GTAGCGTCCC CTTTCGCCCT CGCCGGACAC
  961 GCTGAAACTT GTGGCCGTTT ACGTCGCCCG TCCAGCTCGA CCCAGGATGG GCACCACCCC
 1021 CGGGTGAACA GCTCCTCGCC CCTTGCTCAC CATGAATTTC TGAACCTCAC TCAGATCCTT
1081 CCACCCACCT GACCAACTCT TCCGTGAAGA GGAGAACCCA CCTTTTCCCA AA
//

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original Hisano, Y., Precise in-frame integration of exogenous DNA mediated by CRISPR/Cas9 system in zebrafish. Sci. Rep. 5: 8841 (2015). PMID 25740433. [link to RRC of NBRP]