pEF1a-RoninDHSA-FLAG-IRES-Neo
(Plasmid #28021)

• Depositing Lab: Thomas Zwaka
• Publication: Dejosez et al Genes Dev. 2010 Jul 15. 24(14):1479-84.

Backbone

• Vector backbone: pEF1a-IRES-Neo
• Backbone size w/o insert (bp): 5633
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Ronin
• Alt name: Thap11
• Alt name: pTZ1755
• Species: H. sapiens (human)
• Mutation: DHSA mutation (Y246A) of the previously defined HCF-1-binding motif. By yeast two-hybrid assay, this change completely abolished the ability of Ronin to interact with Hcf-1
• Entrez Gene: THAP11 (a.k.a. HRIHFB2206, CTG-B43a, CTG-B45d, RONIN)
• Tag / Fusion Protein:
  • FLAG (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BglII (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: n/a

Resource Information

• A portion of this plasmid was derived from a plasmid made by: To obtain the pEF1a-Ronin-Flag-Ires-Neo vector, we cloned the PCR product into pEF1-Luc-IRES-Neo (the kind gift of David Spencer, Baylor College of Medicine) using BglII and XbaI, replacing the luciferase gene with Ronin. The DHSA mutation (Y246A) was introduced by site-directed mutagenesis

Terms and Licenses

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

Depositor Comments

Even though the XbaI site is still present, XbaI does not cut this plasmid anymore, possibly due to methylation.