pFB-LIC-Bse
(Plasmid #26108)

• 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
• Depositing Lab: Opher Gileadi
• Publication: Savitsky et al J Struct Biol. 2010 Jun 10. ():.

Backbone

• Vector backbone: pFastBac
• Backbone manufacturer: Invitrogen
• Backbone size (bp): 4803
• Vector type: Insect Expression ; baculovirus expression
• Selectable markers: Gentamicin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Any E. coli cloning host. To generate bacmids, transform DH10Bac according to the manufacturer's procedure (Invitrogen)
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: None
• GenBank ID: EF199842
• Promoter: polyhedrin
• Tag / Fusion Protein:
  • His6 - TEV (N terminal on backbone)

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: fBac-1 (5'-TATTCATACCGTCCCACCA-3')
• 3′ sequencing primer: fBac-2 (5'-GGGAGGTTTTTTAAAGCAAGTAAA-3')

Resource Information

• Supplemental Documents:
  • Vector Information Sheet
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Primers for LIC cloning:

Upstream: add TACTTCCAATCCATG to the 5’ end (ATG in-frame with the desired coding
sequence).

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)