pGagMBPHis
(Plasmid #42176)

• Depositing Lab: James Hurley
• Publication: Carlson et al Proc Natl Acad Sci U S A. 2012 Oct 16;109(42):16928-33. doi: 10.1073/pnas.1211759109. Epub 2012 Oct 1.

Backbone

• Vector backbone: pHIS-parallel.2
• Backbone size w/o insert (bp): 5550
• Total vector size (bp): 8226
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Gag protein sequence is from HIV-1 clone NL4-3
• Species: Synthetic
• Insert Size (bp): 2676
• Tags / Fusion Proteins:
  • MBP (C terminal on insert)
  • 6xHIS (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T7-term

Terms and Licenses

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

Depositor Comments

The Gag protein sequence is from HIV-1 clone NL4-3, GenBank accession no. AAB60571.1, expressed from a codon-optimised synthetic gene (Genscript). 3’ of the synthetic Gag gene are sequences encoding a TEV protease site followed by maltose-binding protein. This GagMBP gene is inserted into the Nde1/Xho1 site of pHis-Parallel2, giving it a C-terminal (His)6 tag for expression of GagMBPHis.