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Please review the QC test results indicated by check icon below as well as clone information before placing your order.

pCAG-EGxxFP

Validation plasmid for the effectiveness of guide RNA in CRISPR/Cas9 system.

Catalog number RDB13532
Resource name pCAG-EGxxFP
Clone info. Validation plasmid for the effectiveness of guide RNA in CRISPR/Cas9 system.
For validation of sgRNA on double strand breaks by observation of green fluorescence. MCS: BamHI, NheI, PstI, SalI, EcoRI, and EcoRV.
Vector backbone pCAGGS (Plasmid)
Selectable markers Amp^r
Gene/insert name Aequorea victoria GFP cDNA
Depositor|Developer Ikawa, Masahito |

Distribution information

Please check terms and conditions set forth by the depositor, which are specified in the RIKEN BRC Catalog and/or Web Catalog.
Terms and conditions set forth by the DEPOSITOR The RECIPIENT of BIOLOGICAL RESOURCE shall obtain a prior written consent on use of it from the DEPOSITOR. 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. In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of the following literature(s) designated by the DEPOSITOR is requested. (Mashiko, D., Sci. Rep. 3: 3355 (2013)) The RECIPIENT which wants to use the BIOLOGICAL RESOURCE even after five (5) years must obtain a written consent from the DEPOSITOR again.
Additional terms and conditions for distribution Regarding resources containing CAG promoter: In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation for the CAG promoter (Niwa, H., Yamamura, K., Miyazaki, J., Gene 108 : 193-200, 1991) and an acknowledgment to Dr. Jun-ich Miyazaki of the Osaka University are requested.
Ordering Please visit Information of Request for Distribution.[link] 
Order form 
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Material Transfer Agreement (MTA for use for not-for-profit academic purpose) [Word]
Schedule A (GFP-A) [Link]
Approval form by depositor [Word]
Remarks ((Additional form)) Approval Form(FormD)
Please use Schedule A (GFP-A) additionally.
Please obtain written approval of the depositor.
Distribution information (domestic users) [open/close]


Catalog # Resource name Shipping form Fee (non-profit org.)
RDB13532 pCAG-EGxxFP DNA solution

Ordering Information [in Japanese] [in English]
Please review the QC test results indicated by check icon below as well as clone information before placing your order.

References and tips

Electronic file

Original reference

original Mashiko, D., Generation of mutant mice by pronuclear injection of circular plasmid expressing Cas9 and single guided RNA. Sci. Rep. 3: 3355 (2013). PMID 24284873.

Further references such as user reports and related articles (go to bottom)

Featured content

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

Sequence information

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 RDB18565_B0Iep1-1.pdf check

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

Primer: pCAG-F
Region: CAG pro,Ensert 5'(EGxxFP-N)
Sequence file: RDB18565_B0Iea.seq check
>13532_18565_B0Ie_2_pCAG-F_H04_22_ABI24.ab1
    1 NNNNNNNNNN NNNNNNNNNN NNNNNNNNNN NNNANGGNGG GGTTCGGCTT CTGGCGNGTG
   61 NNCGGCGGCT CTANCCTCTG CTAACCATGT TCATGCCTTC TTCTTTTTCC TACAGCTCCT
  121 GGGCAACGTG CTGGTTATTG TGCTGTCTCA TCATTTTGGC AAATCTAGAG CCGCCATGGT
  181 GAGCAAGGGC GAGGAGCTGT TCACCGGGGT GGTGCCCATC CTGGTCGAGC TGGACGGCGA
  241 CGTAAACGGC CACAAGTTCA GCGTGTCCGG CGAGGGCGAG GGCGATGCCA CCTACGGCAA
  301 GCTGACCCTG AAGTTCATCT GCACCACCGG CAAGCTGCCC GTGCCCTGGC CCACCCTCGT
  361 GACCACCCTG ACCTACGGCG TGCAGTGCTT CAGCCGCTAC CCCGACCACA TGAAGCAGCA
  421 CGACTTCTTC AAGTCCGCCA TGCCCGAAGG CTACGTCCAG GAGCGCACCA TCTTCTTCAA
  481 GGACGACGGC AACTACAAGA CCCGCGCCGA GGTGAAGTTC GAGGGCGACA CCCTGGTGAA
  541 CCGCATCGAG CTGAAGGGCA TCGACTTCAA GGAGGACGGC AACATCCTGG GGCACAAGCT
  601 GGAGTACAAC TACAACAGCC ACAACGTCTA TATCATGGCC GACAAGCAGA AGAACGGCAT
  661 CAAGGTGAAC TTCAAGATCC GCCACAACAT CGAGGANGGC AGCGTGCAGC TCGCCGACCA
  721 CTACCAGCAG AACACCCCCA TCGGCGANNG CCNCGTGCTG CTGCCCGACA ACCACTGNNG
  781 NNCCGCTAGC CTGCNNNCGA CGAATTCNNT ATCGGCNAGC TGANCCTGAN NTTCATCTGC
  841 ACCANNGGCA ANCTGCCCGN GCCCNNNNNC ACCNNNNNGA NCACCCNNNN NNANNNNNNG
901 CNNNNCTTCA NNNNNNNNNC NNNNNNNNNN
//
Primer: pCAGGS_R1
Region: insert 3'(EGxxFP-C)
Sequence file: RDB18565_B0Ieb.seq check
>13532_18565_B0Ie_2_pCAGGS_R1_H05_23_ABI24.ab1
    1 NNNNNNNNNN ANNCNCTATT GCATGNNNNA AATTTCCTTT ATTAGCCAGA AGTCAGATGC
   61 TCAAGGGGCT TCATGATGTC CCCATAATTT TTGGCAGAGG GAAAAAGATC TCAGTGGTAT
  121 TTGTGAGCCA GGGCATTGGC CACACCAGCC ACCACCTTCT GATAGGCAGC CTGCACCTGA
  181 GGNGTCTCGA GTTACTTGTA CAGCTCGTCC ATGCCGAGAG TGATCCCGGC GGCGGTCACG
  241 AACTCCAGCA GGACCATGTG ATCGCGCTTC TCGTTGGGGT CTTTGCTCAG GGCGGACTGG
  301 GTGCTCAGGT AGTGGTTGTC GGGCAGCAGC ACGGGGCCGT CGCCGATGGG GGTGTTCTGC
  361 TGGTAGTGGT CGGCGAGCTG CACGCTGCCG TCCTCGATGT TGTGGCGGAT CTTGAAGTTC
  421 ACCTTGATGC CGTTCTTCTG CTTGTCGGCC ATGATATAGA CGTTGTGGCT GTTGTAGTTG
  481 TACTCCAGCT TGTGCCCCAG GATGTTGCCG TCCTCCTTGA AGTCGATGCC CTTCAGCTCG
  541 ATGCGGTTCA CCAGGGTGTC GCCCTCGAAC TTCACCTCGG CGCGGGTCTT GTAGTTGCCG
  601 TCGTCCTTGA AGAAGANGGT GCGCTCCTGG ACGTAGCCTT CGGGCATGGC GGACTTGAAG
  661 AAGTCGTGCT GCTTCATGTG GTCGGGGTAG CGGCTGAAGC ACTGCACGCC GTANGTCANG
  721 GTGGTCACGA NGGTGGGCCA GGGCACGGGC AGCTTGCCGG NGGTGCANAT GAACTTCNNG
  781 GTCAGCTTGC CGANATCGAA TTCGTCNANC TGCNNNCTAN CGGNNCCTCA NNGGTTGTCN
  841 GGCAGCAGCA CGGGGNCGTC NCCNANGGGG NNNTTCTGCT GGNNNNNNNC GGNNNNCTNC
901 NNNCTNNNNN NNNNN
//
Primer: EGFP-C-rev
Region: insert mid(EGxxFP-C,MCS,EGxxFP-N)
Sequence file: RDB18565_B0Iec.seq check
>13532_18565_B0Ie_2_EGFP-C-rev_H06_24_ABI24.ab1
    1 NNNNNNNNNN GNNNNTNNCN NNNNGNGGNN NNGGNGCTCA GGTAGTGGTT GTCGGGCAGC
   61 AGCACGGGGC CGTCGCCGAT GGGGGTGTTC TGCTGGTAGT GGTCGGCGAG CTGCACGCTG
  121 CCGTCCTCGA TGTTGTGGCG GATCTTGAAG TTCACCTTGA TGCCGTTCTT CTGCTTGTCG
  181 GCCATGATAT AGACGTTGTG GCTGTTGTAG TTGTACTCCA GCTTGTGCCC CAGGATGTTG
  241 CCGTCCTCCT TGAAGTCGAT GCCCTTCAGC TCGATGCGGT TCACCAGGGT GTCGCCCTCG
  301 AACTTCACCT CGGCGCGGGT CTTGTAGTTG CCGTCGTCCT TGAAGAAGAT GGTGCGCTCC
  361 TGGACGTAGC CTTCGGGCAT GGCGGACTTG AAGAAGTCGT GCTGCTTCAT GTGGTCGGGG
  421 TAGCGGCTGA AGCACTGCAC GCCGTAGGTC AGGGTGGTCA CGAGGGTGGG CCAGGGCACG
  481 GGCAGCTTGC CGGTGGTGCA GATGAACTTC AGGGTCAGCT TGCCGATATC GAATTCGTCG
  541 ACCTGCAGGC TAGCGGATCC TCAGTGGTTG TCGGGCAGCA GCACGGGGCC GTCGCCGATG
  601 GGGGTGTTCT GCTGGTAGTG GTCGGCGAGC TGCACGCTGC CGTCCTCGAT GTTGTGGCGG
  661 ATCTTGAAGT TCACCTTGAT GCCGTTCTTC TGCTTGTCGG CCATGATATA GACGTTGTGG
  721 CTGTTGTAGT TGTACTCCAG CTTGTGCCCC ANGNTGTTGC CGTCCTCCTT NAAGTCGATG
  781 CCCTTCANCT CGATGCGGTT CACCAGGGNG TCGCCCTCNA ACTTCACCTC NGCGCNGGNC
  841 TTGNANTTGC CNNCNNCCTN NAANAAGANN GNGCGCTCCN NNNNNNANCN NTCNGGNCAT
901 NGNNGANTNN NNNNNNNCNN NCNNNTNNNN NNNNNN
//

Please visit Sequencing and PCR primers for primer information.


References

Original, user report and related articles

original Mashiko, D., Generation of mutant mice by pronuclear injection of circular plasmid expressing Cas9 and single guided RNA. Sci. Rep. 3: 3355 (2013). PMID 24284873.
reference Mashiko, D., Feasibility for a large scale mouse mutagenesis by injecting CRISPR/Cas plasmid into zygotes. Dev. Growth Differ. 56 (1): 122-129 (2013). PMID 24372541.

2021.03.11

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