- Backbone size (bp) 5515
Vector typeBacterial Expression
Growth in Bacteria
Growth instructionsFor 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 numberHigh Copy
/ Fusion Protein
- His6 - Z-basic - TEV (N terminal on backbone)
- 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)
Terms and Licenses
Primers for LIC cloning:
Upstream: add TACTTCCAATCCATG to the 5’ end (ATG in-frame with the desired coding
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)
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
Map generated by Addgene from full sequence supplied by depositor.