Purpose(Empty Backbone) Donor vector to generate recombinant baculovirus by site-specific transposition into a baculovirus shuttle vector For use with Bac-to-Bac Expression System insect cells for secreted protein expression
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||26095||Standard format: Plasmid sent in bacteria as agar stab||1||$65 *|
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Backbone manufacturerStructural Genomics Consortium
Vector typeInsect Expression ; Baculovirus expression
- Promoter polyhedrin
/ Fusion Proteins
- His (N terminal on backbone)
- TEV cleavage site (N terminal on backbone)
Growth in Bacteria
Copy numberHigh Copy
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer pFHMSP-fwd (5'-CCGGATTATTCATACCGTCCCACCA-3')
- 3′ sequencing primer pFHMSP-rev (5'-CTGATTATGATCCTCTAGTACTTCT-3') (Common Sequencing Primers)
The pFHMSP-LIC N vector is a derivative of the pFastBac HT A vector (Invitrogen). Honeybee melittin signal sequence was introduced before His tag. It is a donor vector for generation of recombinant baculovirus by site-specific transposition into a baculovirus shuttle vector (bacmid) in E. coli host strain, DH10Bac. For use in Bac-to-Bac Baculovirus Expression System in insect cells for secreted protein expression. This vector adds a 26 amino acid N-terminal fusion tag containing 6X His followed by a TEV cleavage site.
Insertion of DNA sequence into the cloning/expression region is performed using BD-Biosciences Infusion enzyme mediated by directional recombination between complementary nucleotide DNA sequences at the ends of the insert (PCR product) and Nco1/HindIII linearized vector. Insertion of 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:pFHMSP-LIC-N was a gift from Cheryl Arrowsmith (Addgene plasmid # 26095 ; http://n2t.net/addgene:26095 ; RRID:Addgene_26095)