pEGFP-AF8
(Plasmid #117803)

• Purpose: mammalian expression of GFP-tagged human centromere protein A
• Depositing Lab: Kenji Sugimoto
• Publication: Sugimoto et al Cell Struct Funct. 2000 Aug;25(4):253-61.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: CENP-A
• Species: H. sapiens (human)
• Insert Size (bp): 430
• Mutation: See depositor comments below.
• Entrez Gene: CENPA (a.k.a. CENP-A, CenH3)
• Tag / Fusion Protein:
  • GFP (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SacI (not destroyed)
• 3′ cloning site: XhoI/SalI (destroyed during cloning)
• 5′ sequencing primer: CMV-F

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The insert fragment with Sac I and Xho I sites was cloned into Sac I and Sal I sites of pEGFP-C1.
Thus, there is no Xho I and Sal I site in pEGFP-AF8.
The stop codon was also changed to TCG (..ctcggcTCGac-3’)
Note that the cloning strategy adds additional amino acids onto the C-terminus of the insert (STVPRARDPPDLDN), but this does not interfere with the function.