pQE-80L MBP-SspB Micro
(Plasmid #60410)

• Purpose: E. coli expression of SspB Micro
• Depositing Lab: Brian Kuhlman
• Publication: Guntas et al Proc Natl Acad Sci U S A. 2015 Jan 6;112(1):112-7. doi: 10.1073/pnas.1417910112. Epub 2014 Dec 22.

Backbone

• Vector backbone: pQE-80L
• Backbone manufacturer: Qiagen
• Backbone size w/o insert (bp): 5979
• Total vector size (bp): 6315
• Modifications to backbone: Added N-terminal MBP
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: SspB micro
• Species: E. Coli
• Insert Size (bp): 336
• Mutation: R73Q, Y11K and A15E
• Tag / Fusion Protein:
  • His-MBP-TEV (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamH1 (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: pQE-80L Fwd
• 3′ sequencing primer: pQE-80L Rev

Resource Information

• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor Comments

The SspB used in this construct contains two amino acid mutations that disrupt the natural dimerization of SspB. These mutations are Y11K and A15E (residue numbering from pdb code 1ou9). Here is the sequence around the mutated residues with the sites of mutation in parentheses: SSPKRP(K)LLR(E)YYDWLVDNS