Purpose(Empty Backbone) SGC Baculovirus transfer vector with His tag in 22-aa N-terminal fusion peptide with TEV protease cleavage site. Includes sites for LIC cloning, and SacB gene for negative selection
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||26108||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 4803
Vector typeInsect Expression ; baculovirus expression
Growth in Bacteria
Growth instructionsAny E. coli cloning host. To generate bacmids, transform DH10Bac according to the manufacturer's procedure (Invitrogen)
Copy numberHigh Copy
- Promoter polyhedrin
/ Fusion Protein
- His6 - TEV (N terminal on backbone)
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer fBac-1 (5'-TATTCATACCGTCCCACCA-3')
- 3′ sequencing primer fBac-2 (5'-GGGAGGTTTTTTAAAGCAAGTAAA-3') (Common Sequencing Primers)
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).
Generate bacmids and baculoviruses using the Bac to Bac method (Invitrogen)
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:pFB-LIC-Bse was a gift from Opher Gileadi (Addgene plasmid # 26108 ; http://n2t.net/addgene:26108 ; RRID:Addgene_26108)
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