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pET28/His6-mCherry-D4

Marker to localize cholesterol-rich domain of the plasma membrane. Expression vector of His and mCherry-tagged C. perfringens perfringolysin theta derivatives encoding the D4 for E. coli.

Catalog number RDB14300
Resource name pET28/His6-mCherry-D4
Clone info. Expression vector of His and mCherry-tagged C. perfringens perfringolysin theta derivatives encoding the D4 (domain 4 of theta toxin; K363-N472) for E. coli, see Fig. 1A of Shimada, Y. et. al. Eur. J. Biochem, 269: 6195-6203, 2002.
Expression was confirmed by the depositor with SDS-PAGE. DQ673099.1 for Clostridium perfringens CN3947 perfringolysin O (pfoA) gene.
Vector backbone pET-28b (Plasmid)
Size of vector backbone 5.4 kb
Selectable markers Kan^r
Gene/insert name Clostridium perfringens perfringolysin O (pfoA) cDNA
Depositor Sako, Yasushi | Abe, Mitsuhiro |

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 literature designated by the DEPOSITOR is requested. (Makino, A. et al., FASEB J. 2016 Aug 4. pii: fj.201500075R.)
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Remarks ((Remarks)) Please contact us. We will provide you with appropriate documents.
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Catalog # Resource name Shipping form Fee (non-profit org.)
RDB14300 pET28/His6-mCherry-D4 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 RDB14300_A6Hup1.pdf

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

Primer: T7_20bp RDB14300_A6Hua.seq
>D03762A1_A6Hu_1_T7_20bp_G10_19_ABI24.ab1
    1 TCGGGTCATT CCGTCTAGAA TAATTTTGTT TACTTTAAGA AGGAGATATA CCATGGGCAG
   61 CAGCCATCAT CATCATCATC ACAGCAGCGG CCTGGTGCCG CGCGGCAGCG GTACCATGGT
  121 GAGCAAGGGC GAGGAGGATA ACATGGCCAT CATCAAGGAG TTCATGCGCT TCAAGGTGCA
  181 CATGGAGGGC TCCGTGAACG GCCACGAGTT CGAGATCGAG GGCGAGGGCG AGGGCCGCCC
  241 CTACGAGGGC ACCCAGACCG CCAAGCTGAA GGTGACCAAG GGTGGCCCCC TGCCCTTCGC
  301 CTGGGACATC CTGTCCCCTC AGTTCATGTA CGGCTCCAAG GCCTACGTGA AGCACCCCGC
  361 CGACATCCCC GACTACTTGA AGCTGTCCTT CCCCGAGGGC TTCAAGTGGG AGCGCGTGAT
  421 GAACTTCGAG GACGGCGGCG TGGTGACCGT GACCCAGGAC TCCTCCCTGC AGGACGGCGA
  481 GTTCATCTAC AAGGTGAAGC TGCGCGGCAC CAACTTCCCC TCCGACGGCC CCGTAATGCA
  541 GAAGAAGACC ATGGGCTGGG AGGCCTCCTC CGAGCGGATG TACCCCGAGG ACGGCGCCCT
  601 GAAGGGCGAG ATCAAGCAGA GGCTGAAGCT GAAGGACGGC GGCCACTACG ACGCTGAGGT
  661 CAAGACCACC TACAAGGCCA AGAAGCCCGT GCAGCTGCCC GGCGCCTACA ACGTCAACAT
  721 CAAGTTGGAC ATCACCTCCC ACAACGAGGA CTACACCATC GTGGAACAGT ACGAACGCGC
  781 CGAGGGCCGC CACTCCACCG GCGGCATGGA CGAGCTGTAC AAGAAGCTTA AGGGAAAAAT
  841 AAACTTAGAT CATAGTGGAG CCTATGTTGC ACAGTTTGAA GTAGCCTGGG ATGAAGTTTC
  901 ATATGACAAA GAAAGAAATG AAGTTTTAAC TCATAAAACA TGGGATGGAA ATTATCAAGA
  961 TAAAACAGCT CACTATTCAA CAGTAATACC TCTTGAAGCT AATGCAAGAA TATAGAATAA
 1021 AAGCCAGAGA GTGTACAGGC CTTGCTTGGG GAATGGTGGA GAGATGGTAT AAGTGATATG
 1081 ATGTTCCCAT TAACCAATGT ATGATGTTTC AATATGGGGA ACCAACTTAA TACCCCTTGG
1141 AC
//
Primer: T7_terminator RDB14300_A6Hub.seq
>D03762A1_A6Hu_1_T7 Terminator_H10_22_ABI24.ab1
    1 CCTTAGCCTT TCTTTTCGGG GCTTTGTTAG CAGCCGGATC TCAGTGGTGG TGGTGGTGGT
   61 GCTCGAGTTA ATTGTAAGTA ATACTAGATC CAGGGTATAA AGTTGTTCCC CATATTGAAA
  121 CATTTATATT ATTTGTTAAT GGAACATCAT ATTCACTTAT AACATCTCTC CACCATTCCC
  181 AAGCAAGGCC TGTACACTCT CTTGCTTTTA TTCTTATATT TCTTGCATTA GCTTCAAGAG
  241 GTATTACTGT TGAATAGTGA GCTGTTTTAT CTTGATAATT TCCATCCCAT GTTTTATGAG
  301 TTAAAACTTC ATTTCCTTCT TTGTCATATG AAACTTCATC CCAGGCTACT TCAAACTGTG
  361 CAACATAGGC TCCACTATGA TCTAAGTTTA TTTTTCCCTT AAGCTTCTTG TACAGCTCGT
  421 CCATGCCGCC GGTGGAGTGG CGGCCCTCGG CGCGTTCGTA CTGTTCCACG ATGGTGTAGT
  481 CCTCGTTGTG GGAGGTGATG TCCAACTTGA TGTTGACGTT GTAGGCGCCG GGCAGCTGCA
  541 CGGGCTTCTT GGCCTTGTAG GTGGTCTTGA CCTCAGCGTC GTAGTGGCCG CCGTCCTTCA
  601 GCTTCAGCCT CTGCTTGATC TCGCCCTTCA GGGCGCCGTC CTCGGGGTAC ATCCGCTCGG
  661 AGGAGGCCTC CCAGCCCATG GTCTTCTTCT GCATTACGGG GCCGTCGGAG GGGAAGTTGG
  721 TGCCGCGCAG CTTCACCTTG TAGATGAACT CGCCGTCCTG CAGGGAGGAG TCCTGGGTCA
  781 CGGTCACCAC GCCGCCGTCC TCGAAGTTCA TCACGCGCTC CCACTTGAAG CCCTCGGGGA
  841 AGGACAGCTT CAAGTAGTCG GGGATGTCGG CGGGGTGCTT CACGTAGGCC TTGGAGCCGT
  901 ACATGAACTG AGGGGACAGG ATGTCCCAGG CGAAGGGCAG GGGGCCACCC TTGGTCACCT
  961 TCAGCTTGGC GGTCTGGGTG CCCTCGTAAG GGGCGGCCCT CGCCCTCGCC CTCGATCTCG
 1021 AACTCGTGGC CGTTCACGGA GCCCTCCATG TGCACCTTGA GCGCATGAAA CCTCCTTTGA
 1081 TGATTGCATG TATCTCCTCG CCATGCCTCA CATGGTACGG CTGGCCGCGC GCAACCAGCC
1141 GCCTTGCTGT GGAATGAATG AAAGGA
//

Please visit plasmidSequencing and PCR primers for primer information.

References and tips

Electronic file

Featured content

Featured content Dnaconda's recommendation EXR0062j (Japanese text)
Featured content Sphingolipid signaling pathway (English text)

References

original Makino, A., A novel sphingomyelin/cholesterol domain-specific probe reveals the dynamics of the membrane domains during virus release and in Niemann-Pick type C. FASEB J. 2016 Aug 4. pii: fj.201500075R. [Epub ahead of print] PMID 27492925.

2017.09.11

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