pFHMSP-LIC-C
(Plasmid #26100)

• 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• Publication: Cloning Vectors (unpublished)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-C
• Backbone manufacturer: Structural Genomics Consortium
• Vector type: Insect Expression ; Baculovirus expression
• Promoter: polyhedrin
• Tags / Fusion Proteins:
  • 8x His tag (C terminal on backbone)
  • MKFLVNVALVFMVVYISYIYAAA (Honeybee melittin signal sequence) (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-C
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

The pFHMSP-LIC C vector is a derivative of the pFastBac HT A vector (Invitrogen). His tag was replaced by a Honeybee melittin signal sequence and His tag placed into C-term. 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 one or two (A) amino acid after releasing signal peptide by signalase.

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 Not1/EcoR1
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