pFBOH-MHL
(Plasmid #62304)

• Purpose: (Empty Backbone) It is a donor vector for generation of recombinant baculovirus by site-specific transposition in an E. coli host.
• Depositing Lab: Cheryl Arrowsmith
• Publication: Arrowsmith Lab Plasmids (unpublished)

Backbone

• Vector backbone: pFBOH-MHL
• Backbone manufacturer: Structural Genomics Consortium
• Backbone size (bp): 6767
• Vector type: Baculovirus expression
• Promoter: polyhedrin
• Tags / Fusion Proteins:
  • 6xHis (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
• Growth instructions: Gentamicin (bacmid resistance in DH10Bac E. coli)
• Copy number: Unknown

Cloning Information

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

Resource Information

• Supplemental Documents:
  • pFBOH-MHL
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

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.