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pmKO-AF8
(Plasmid #118224)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 118224 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pmKO1-MC1
  • Backbone manufacturer
    MBL, Nagoya, Aichi, Japan
  • Backbone size w/o insert (bp) 4656
  • Vector type
    Mammalian Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    CENP-A
  • Alt name
    centromere protein A
  • Species
    H. sapiens (human)
  • Entrez Gene
    CENPA (a.k.a. CENP-A, CenH3)
  • Promoter CMV
  • Tag / Fusion Protein
    • Kusabira orange (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BglII/BamHI (destroyed during cloning)
  • 3′ cloning site KpnI (not destroyed)
  • 5′ sequencing primer CMV-F
  • 3′ sequencing primer SV40pA-R
  • (Common Sequencing Primers)

Resource Information

  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry

Depositor Comments

Bgl II-KpnI fragment of pEGFP-AF8 (Addgene plasmid # 117803) was recloned into BamHI and KpnI site of pmKO1-MC1.

Note that the cloning strategy adds additional amino acids onto the C-terminus of the insert (STVPELQQRIRETREQSLNKRPRL), but this does not interfere with plasmid function. The extra sequence includes an MCS for adding tags to the C-terminus of the insert.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pmKO-AF8 was a gift from Kenji Sugimoto (Addgene plasmid # 118224 ; http://n2t.net/addgene:118224 ; RRID:Addgene_118224)
  • For your References section:

    Construction of three quadruple-fluorescent MDA435 cell lines that enable monitoring of the whole chromosome segregation process in the living state. Sugimoto K, Senda-Murata K, Oka S. Mutat Res. 2008 Nov 17;657(1):56-62. doi: 10.1016/j.mrgentox.2008.08.005. Epub 2008 Aug 19. 10.1016/j.mrgentox.2008.08.005 PubMed 18778791