pKM1000
(Plasmid #8840)

• Purpose: (Empty Backbone)
• Depositing Lab: David Waugh
• Publication: Fox et al FEBS Lett. 2003 Feb 27. 537(1-3):53-7.

Backbone

• Vector backbone: pET11
• Backbone manufacturer: Novagen
• Backbone size (bp): 6700
• Modifications to backbone: DNA encoding presumptive N-terminal signal peptide deleted. The single cysteine in T. maritima MBP (residue 12) was changed to a serine by site-directed mutagenesis.
• Vector type: Bacterial Expression
• Tag / Fusion Protein:
  • T. maritima MBP (N terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): ccdB Survival
• Growth instructions: Because it carries a gene (ccdB ) that encodes a lethal DNA gyrase poison, this vector must be propogated in an E. coli gyrA mutant such as DB3 or DB5 (Invitrogen), which is immune to the action of CcdB.
• Copy number: Low Copy

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: pBRrevBam; T7
• 3′ sequencing primer: T7term

Terms and Licenses

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

Depositor Comments

A Gateway destination vector for the production of recombinant proteins as fusions to the C-terminus of T. maritima maltose-binding protein. DNA encoding presumptive N-terminal signal peptide deleted. The single cysteine in T. maritima MBP (residue 12) was changed to a serine by site-directed mutagenesis.