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.

pCMV-3xFLAG-mGli3_del97G-HA (#RDB15041)

Expression vector of mouse mutant Gli3, one-base-pair deletion of 97G, tagged with N-terminal 3xFLAG and C-terminal HA. Though this clone was constructed based on the pEGFP-C1, GFP was removed.

Clone info. Expression vector of mouse mutant Gli3, one-base-pair deletion of 97G, tagged with N-terminal 3xFLAG and C-terminal HA, see Fig3A of Makino, S. et al. Sci. Rep., 6: 39608, 2016.
Comment Expression was confirmed by the depositor with direct sequencing and western blotting.
Vector backbone
pEFGP-C1 (plasmid)
Selectable markers Kan^r (E. coli), Neo^r (mammalian cells).
Gene/insert name mouse Gli3 cDNA
Depositor|Developer Gondo, Yoichi | Makino, Shigeru |
 

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 ]
MTA, for use for not-for-profit academic purpose [Word] [Example of MTA ]
Please visit Information of Request for Distribution.[link] For for-profit-research purpose, please contact us. 
Terms and conditions for distribution In publishing the research results obtained by use of the BIOLOGICAL RESOURCE, a citation of literature designated by the DEPOSITOR is requested (Makino,S. et al. Sci. Rep., 6: 39608, 2016).
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 ]
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 (Makino,S. et al. Sci. Rep., 6: 39608, 2016)。

Catalog # Resource name Shipping form Fee (non-profit org.)
RDB15041 pCMV-3xFLAG-mGli3_del97G-HA 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 pCMV-3xFLAG-mGli3_del97G-HA was provided by the RIKEN BRC through the National BioResource Project of the MEXT, Japan (cat. RDB15041).

Reference section:

Makino, S., Fukumura, R., Gondo ,Y., Illegitimate translation causes unexpected gene expression from on-target out-of-frame alleles created by CRISPR-Cas9. Sci. Rep. 6: 39608 (2016). PMID 28000783. [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 RDB15041_A7Eop1-1.pdf check

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

Primer: CMV_Forward (Pr0016)
Region: CMV pro, 3xFLAG, insert 5' (del97G)
Sequence file: RDB15041_A7Eoa.seq check
>D04539A5_A7Eo_1_CMV-Forword_G02_20_ABI24.ab1
    1 AGTCAACTCC GCCCATTGAC GCAATGGGCG GTAGGCGTGT ACGGTGGGAG GTCTATATAA
   61 GCAGAGCTGG TTTAGTGAAC CGTCAGATCC GCTAGCACCA TGGACTACAA AGACCATGAC
  121 GGTGATTATA AAGATCATGA CATCGATTAC AAGGATGACG ATGACAAGGA GGCCCAGGCC
  181 CACAGCTCTA CGGCGACTGA GAGGAAGAAA GCTGAAAATT CCATTGGGAA ATGTCCCACG
  241 AGAACAGATG TCAGCGAGAA GCCGTGGCCT CTAGTACCAC TTCCAATGAG GATGAAAGTC
  301 CTGGACAGAT CTATCACCGA GAGAGAAGAA ACGCAATCAC TATGCAGCCT CAGAGTGTGC
  361 AGGGTCTCAA CAAAATCAGT GAGGAGCCCT CGACGTCTAG TGATGAGAGG GCCTCGCTGA
  421 TCAAGAAAGA GATCCATGGC TCTCTACCAC ATCTGGCGGA GCCCTCTCTC CCTTACCGTG
  481 GGACTGTGTT TGCCATGGAT CCCCGGAATG GCTACATGGA GCCTCACTAC CACCCTCCTC
  541 ATCTTTTCCC TGCCTTCCAT CCTCCTGTAC CAATTGATGC CAGACATCAT GAGGGCCGTT
  601 ACCATTATGA TCCATCTCCT ATTCCTCCAT TACATGTGCC TTCTGCCTTA TCTAGTAGCC
  661 CGACGTATCC AGACTTGCCC TTCATTAGGA TCTCCCCACA CCGTAATCCC ACTGCAGCCT
  721 CAGAGTCCCC CTTCAGCCCC CCACACCCCT ACATCAACCC ATATATGGAC TACATCCGCT
  781 CATTGCACAG CAGCCCATCC CTCTCCATGA TCTCTGCTGC CCGAGGGCTG AGCCCTACAG
  841 ATGCTCCCCA TGCTGGAGTC AGCCCTGCGG AATACTATCA CCAGATGGCT CTGCTGACAG
  901 GCCAGCGCAG CCCTTATGCA GACATCCTTC CCTCAGCTGC CACTGCTGGT GCACGGGCCA
  961 TCCATATGGA GTACCCTTCA TGCCATGGAC AGCACCAGAT TTCCCAGCCC TAGCTGTCAG
 1021 CTAGCCAGGC CGGAAGCGCA CACTGTCATA TCGCCACTGT CAGATCCATA GCCTTCCGAC
1081 ATTCGACCAT GATAGACATC CCCCTACTCT GGTTTAACAA TTCCCTCCAA T
//
Primer: SV40polyA-R (Pr0109)
Region: HA, insert 3'
Sequence file: RDB15041_A7Eob.seq check
>D04539A5_A7Eo_1_SV40polyA-R_H02_23_ABI24.ab1
    1 AAAAGTGGGA GTTTTAGCAG TAAACCTCTA CAAATGTGGT ATGGCTGATT ATGATCAGTT
   61 ATCTAGATCC GGTGGATCCC GGGCCCGCGG TACCGTCGAC TGCAGAATTC CTATTAAGCG
  121 TAATCTGGAA CATCGTATGG GTAGGCCTGC ATAACTGCAA GGAACTTGCT TTCTTCTGCA
  181 AGGGAGGTCA GCAAAGAACT CATATCCCCG ATAGCCATGT TGGTGGTGCC CATGGATAGG
  241 GATGGGAATG GGAGGGATGC CCGCGGAGTG GTGAGACGGG AGGAGCTGTG GGAAAGGTTC
  301 TGAATAATAC TTGGGCTCAA GGCCCCTGAC ATTAGGCTGG TATGGTCCCC ATCATCTATG
  361 ATGGCATCAA AATCAATCTG CACACCTTCT AGGTCATGAC TCTCAAAGCT GTCAACTGTG
  421 CTTGTTACCT GGTTAGCACC TGGGGAGAGT AACTCAGAGT TTTCAGTGCC ATTCCCTTGG
  481 AGCAGGCAGT TGGCATCTGA GAGGGAGAAC GAGCCAGCTT TCCTGTCTTG CTGACTGAAG
  541 CCCATGGTTT GGTCATAGAA CTGACCGGAA TAATTCTGCA CGGTAGAGCA GAGCTGCTGG
  601 GACTGACCTT GTGCCTCCAT TTTGATGCTG TTCACCCTGC TGCTCTGACT CATGTCACTG
  661 AGCTGTCCTT GTTGCAGAGT GAGGTTGCCC CTTGGCATTG CCTGACGCCT GTTACCCCCA
  721 TAGCTGGCAC AGGGCTGGTA GCCCCTGATA GCTGCCATGC TTGATGGCTG GCTGCTGTTG
  781 TCCTGAGTCC CTGGTAGGCA GCTCTCTGTC CCTTGATAGA TGTTGATATG TGAGGTAGCA
  841 CCAACCTGCC CTAGTACCTG TTGACCAGGG CGGCTGGGCC CCTGGTGTTG CATGCTGCCA
  901 CTGAGTAGCT GATGGGCAAT GTATCCCTGT CCCATTTGAT CTTGGGTTTG CATGCCATTC
  961 ACTGTGTTGC CAGCTAACTC ATTTGGTGCC ACGGGACAGG CTGAAATCTA GCAACCCCCA
 1021 TTCTCTTGCA AGCTGGTGCG TTTAGCTGAC CCCTTGAAAT CCCTGTACAC AGCCATGATC
 1081 TAGAGCACTG GAGTCATTGT CTGGCCTGTG GATCTGACTC GATCACTGCT GCTCAAGGGC
1141 TGATTCCATG
//
Primer: SV40pro_ori_F (Pr0731)
Region: SV40pro_ori, NeoR_KanR
Sequence file: RDB15041_A7Eoc.seq check
>15041_15041_A9IbF_SV40pro_ori_F_F06_18_ABI24.ab1
    1 NNNNNNNNNN NNNGCNGCCT CGNCCTCTGN NCTATTCNNG AAGTAGTGAG GAGGCTTTTT
   61 TGGAGGCCTA GGCTTTTGCA AAGATCGATC AAGAGACAGG ATGAGGATCG TTTCGCATGA
  121 TTGAACAAGA TGGATTGCAC GCAGGTTCTC CGGCCGCTTG GGTGGAGAGG CTATTCGGCT
  181 ATGACTGGGC ACAACAGACA ATCGGCTGCT CTGATGCCGC CGTGTTCCGG CTGTCAGCGC
  241 AGGGGCGCCC GGTTCTTTTT GTCAAGACCG ACCTGTCCGG TGCCCTGAAT GAACTGCAAG
  301 ACGAGGCAGC GCGGCTATCG TGGCTGGCCA CGACGGGCGT TCCTTGCGCA GCTGTGCTCG
  361 ACGTTGTCAC TGAAGCGGGA AGGGACTGGC TGCTATTGGG CGAAGTGCCG GGGCAGGATC
  421 TCCTGTCATC TCACCTTGCT CCTGCCGAGA AAGTATCCAT CATGGCTGAT GCAATGCGGC
  481 GGCTGCATAC GCTTGATCCG GCTACCTGCC CATTCGACCA CCAAGCGAAA CATCGCATCG
  541 AGCGAGCACG TACTCGGATG GAAGCCGGTC TTGTCGATCA GGATGATCTG GACGAAGAGC
  601 ATCAGGGGCT CGCGCCAGCC GAACTGTTCG CCAGGCTCAA GGCGAGCATG CCCGACGGCG
  661 AGGATCTCGT CGTGACCCAT GGCGATGCCT GCTTGCCGAA TATCATGGTG GAAAATGGCC
  721 GCTTTTCTGG ATTCATCGAC TGTGGCCGGC TGGGTGTGGC GGANCGCTAT CANGACATAG
  781 CGTTGGCTAC CCGTGATATT GCTGAAGANC TTGGCGGCGA ATGNGCTGAC CGCTTCCTCG
  841 TGCTTTACGG TATCGCCGCT CCCGATTCGC AGCGCATCGC NNTCTATCGC CTTCTTGNNG
  901 AGTNCTTCTG ANCGGGNNTC NGGGGGTTCN AAATGANNGA NNNANCGANN NNNNANNNGN
961 NNTCNNNANN TTNNNATTNN NNNGNNNNNC NTNNNNNNNA NNNNNNG
//

Please visit Sequencing and PCR primers for primer information.

References

Original, user report and related articles

original Makino, S., Illegitimate translation causes unexpected gene expression from on-target out-of-frame alleles created by CRISPR-Cas9. Sci. Rep. 6: 39608 (2016). PMID 28000783. [link to RRC of NBRP]