pICE-HA-Ku70-siR-Mut6E
(Plasmid #82330)

• Purpose: Plasmid for constitutive or doxy-inducible expression of mutant human Ku70 unable to bind DNA and resistant to siRNA. Confers resistance to puro. Use T-REx cells for doxycycline-inducible expression.
• Depositing Labs: Sébastien Britton, Patrick Calsou
• Publication: Lamaa et al EMBO Rep. 2016 Apr;17(4):508-18. doi: 10.15252/embr.201541181. Epub 2016 Mar 10.

Backbone

• Vector backbone: pICE
• Backbone manufacturer: Synthetic - Addgene plasmid #46960
• Backbone size w/o insert (bp): 5009
• Total vector size (bp): 6844
• Modifications to backbone: HA tag and cloning sites
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

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: Ku70
• Alt name: XRCC6
• Species: H. sapiens (human)
• Insert Size (bp): 1830
• Mutation: Mut6E corresponding to K282E K287E T300E K331E K338E R403E; mutant unable to bind to DNA
• Entrez Gene: XRCC6 (a.k.a. CTC75, CTCBF, G22P1, KU70, ML8, TLAA)
• Promoter: CMV-tet
• Tag / Fusion Protein:
  • HA (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-R

Resource Information

• Supplemental Documents:
  • Author annotated GenBank sequence

Terms and Licenses

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