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pNIC-ZB
(Plasmid #26107)

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Available to Academic and Nonprofits Only

Backbone

  • Vector backbone
    pET-28-a
  • Backbone manufacturer
    Novagen
  • Backbone size (bp) 5515
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    For plasmid propagation and cloning, use any E. coli strain that does not express T7 RNA polymerase. For expression, use host cells that express T7 RNA pol, e.g. BL21(DE3).
  • Copy number
    High Copy

Sequence Information

Gene/Insert

  • Gene/Insert name
    None
  • GenBank ID
    GU452710
  • Tag / Fusion Protein
    • His6 - Z-basic - TEV (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site Ligation-Independent cloning; cut with BsaI (destroyed during cloning)
  • 3′ cloning site Ligation-Independent cloning; cut with BsaI (destroyed during cloning)
  • 5′ sequencing primer T7F
  • 3′ sequencing primer T7R
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Primers for LIC cloning:

Upstream: add TACTTCCAATCCATG to the 5’ end (ATG in-frame with the desired coding
sequence).

Downstream: add TATCCACCTTTACTG to 5’ end of downstream primer; add termination
codon, if necessary.

Detailed cloning method available in the paper (Savitsky et al., J Struct Biol., 2010)

How to cite this plasmid

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:

    pNIC-ZB was a gift from Opher Gileadi (Addgene plasmid # 26107)
  • For your References section:

    High-throughput production of human proteins for crystallization: The SGC experience. Savitsky P, Bray J, Cooper CD, Marsden BD, Mahajan P, Burgess-Brown NA, Gileadi O. J Struct Biol. 2010 Jun 10. ():. 10.1016/j.jsb.2010.06.008 PubMed 20541610