pmKO-AF8
(Plasmid #118224)

• Purpose: Kusabira- Orange-CENP-A expression plasmid
• Depositing Lab: Kenji Sugimoto
• Publication: Sugimoto et al Mutat Res. 2008 Nov 17;657(1):56-62. doi: 10.1016/j.mrgentox.2008.08.005. Epub 2008 Aug 19.

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, 50 μg/mL
• 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

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• 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.