pKSJ155
(Plasmid #27126)

• Depositing Lab: Alan Finkelstein
• Publication: Kienker et al J Gen Physiol. 2000 Oct . 116(4):587-98.

Backbone

• Vector backbone: pET15b
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5708
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL10 Gold
• Growth instructions: The plasmid is provided in XLGold (Stratagene), a non-expression (recA) strain for plasmid preparation. Use BL21 (DE3) or similar strain for expression from pET vectors
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Colicin Ia + Ia immunity protein
• Species: E. coli
• Tag / Fusion Protein:
  • His6 (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: BamHI (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

This plasmid contains the C-terminal channel forming domain of colicin Ia and the Ia immunity protein. The construct is also referred to as "CT-Medium or CT-M".

The SOS promoter for colicin expression is at around residues 148-165 of the insert, in the forward direction. The immunity promoter -35 box is about 460 bp from the end of the insert; there are only about 20 bp between the terminator for the colicin gene, reading in one direction, and the terminator for the immunity protein, reading in the other.