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pCMV-3xFLAG-mGli3_del97G-HA

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.

Catalog number RDB15041
Resource name pCMV-3xFLAG-mGli3_del97G-HA
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.
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 Gondo, Yoichi | Makino, Shigeru |
 
Other clones in our bank mouse Gli3 (NCBI Gene 14634) |

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 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).
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RDB15041 pCMV-3xFLAG-mGli3_del97G-HA DNA solution

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References and tips

Electronic file

Original reference

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.

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

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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 RDB15041_A7Eop1-1.pdf

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

Region: CMV pro, 3xFLAG, insert 5' (del97G) | Primer: CMV_Forward | Sequence: RDB15041_A7Eoa.seq
>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
//
Region: HA, insert 3' | Primer: SV40polyA-R | Sequence: RDB15041_A7Eob.seq
>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
//
Region: SV40pro_ori, NeoR_KanR | Primer: SV40pro_ori_F | Sequence: RDB15041_A7Eoc.seq
>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.

2019.10.10

GNP_filter3_RDBDEP_html_190927.pl