|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||62304||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerStructural Genomics Consortium
- Backbone size (bp) 6767
Vector typeBaculovirus expression
- Promoter polyhedrin
/ Fusion Proteins
- 6xHis (N terminal on backbone)
- TEV cleavage site (N terminal on backbone)
Growth in Bacteria
Growth instructionsGentamicin (bacmid resistance in DH10Bac E. coli)
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer pFBOH-fwd (5'-CCGGATTATTCATACCGTCCCACCA-3')
- 3′ sequencing primer pFBOH-rev (5'-CTGATTATGATCCTCTAGTACTTCT-3') (Common Sequencing Primers)
The pFBOH-MHL vector is a derivative of the pFBOH-LIC vector (SGC). It is a donor vector for generation of recombinant baculovirus by site-specific transposition in an E. coli host. This vector adds an 18 amino acid N-terminal fusion tag containing a 6X His followed by a TEV cleavage site. Two stop codons are included in the vector at the C-terminal cloning site.
Insertion of a DNA sequence into the cloning/expression region is performed using Clontech’s In-fusion enzyme mediated directional recombination between complementary 15 nucleotide DNA sequences at the ends of the insert (PCR product) and BseRI linearized vector. Insertion of a target sequence involves replacement of a SacB gene stuffer sequence, which provides for negative selection of the original plasmid on 5% sucrose.
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:pFBOH-MHL was a gift from Cheryl Arrowsmith (Addgene plasmid # 62304 ; http://n2t.net/addgene:62304 ; RRID:Addgene_62304)