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pCLR2070
(Plasmid #28063)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 28063 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

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

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • 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
  • (Common Sequencing Primers)

Resource Information

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)

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pCLR2070 was a gift from Keith Joung (Addgene plasmid # 28063 ; http://n2t.net/addgene:28063 ; RRID:Addgene_28063)
  • For your References section:

    An improved zinc-finger nuclease architecture for highly specific genome editing. Miller JC, Holmes MC, Wang J, Guschin DY, Lee YL, Rupniewski I, Beausejour CM, Waite AJ, Wang NS, Kim KA, Gregory PD, Pabo CO, Rebar EJ. Nat Biotechnol. 2007 Jul . 25(7):778-85. 10.1038/nbt1319 PubMed 17603475