pCLR2070
(Plasmid #28063)

• Depositing Lab: Keith Joung
• Publication: Miller et al Nat Biotechnol. 2007 Jul . 25(7):778-85.

Backbone

• Vector backbone: pMLM292
• Backbone size w/o insert (bp): 5725
• Vector type: Mammalian Expression ; Zebrafish expression

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: FokI endonuclease
• Mutation: Contains N496D point mutation in FokI domain, as well as Q486E and I499L present in the vector backbone
• Tag / Fusion Protein:
  • FLAG (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: CMV-F

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

For expression of a ZFN targeted to a site with a 5 or 6 bp spacer and “-” ELD heterodimeric FokI domain. (Doyon et. al,Nat Biotech 2011) in mammalian cells or zebrafish as described by Moore et al., PLoS One 2012 (https://www.ncbi.nlm.nih.gov/pubmed/22655075) and Cade et al., Nucleic Acids Res 2012 (https://www.ncbi.nlm.nih.gov/pubmed/22684503)