pFHMSP-LIC-N
(Plasmid #26095)

• 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
• Depositing Lab: Cheryl Arrowsmith
• Publication: Cloning Vectors (unpublished)

Backbone

• Vector backbone: pFHMSP-LIC-N
• Backbone manufacturer: Structural Genomics Consortium
• Vector type: Insect Expression ; Baculovirus expression
• Promoter: polyhedrin
• Tags / Fusion Proteins:
  • His (N terminal on backbone)
  • TEV cleavage site (N terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: pFHMSP-fwd (5'-CCGGATTATTCATACCGTCCCACCA-3')
• 3′ sequencing primer: pFHMSP-rev (5'-CTGATTATGATCCTCTAGTACTTCT-3')

Resource Information

• Supplemental Documents:
  • pFHMSP-LIC-N

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Industry MTA

Depositor Comments

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.

http://www.sgc.utoronto.ca/SGC-WebPages/toronto-vectors.php