Part:BBa_K1962002

Truncated Colicin Ia Lacking Bacteriocin Active Domain

Colicin Ia is a bacteriocin secreted by E. coli. Colicins typically have 3 domains, a translocation domain, a receptor binding domain, and a cytotoxic domain. If you are looking for a full-length Colicin Ia BioBrick it is located here BBa_K1962000.

This part encodes a truncated version of Colicin Ia that lacks the C-terminal bacteriocin domain. The part has an engineered multiple cloning site at the 3' end, preceding the RFC[10] suffix and regulation double stop codons, containing the following in-frame restriction sites: BamHI, KpnI, SalI, BglII and NheI. The presence of this multiple cloning site will allow for different toxic domains to be fused at the C-terminus of the truncated colicin and thereby generating a synthetic colicin in a rapid and facile manner.

Usage and Biology We have also cloned an antibacterial toxin, known as ssp2 which is a substrate of the T6SS in serratia marcescens onto the C-terminal of this colicin and submitted it as a Biobrick (BBa). This was to create a synthetic colicin unrecognisable to our target pathogens, E. coli and Salmonella.   Below is the structure of Colicin iA, in green is the receptor binding domain, in red the translocation domain and in blue the cytotoxic domain. We have removed the cytotoxic domain and replaced it with a multiple cloning site. This was then cloned into pSB1C3 with the Biobrick prefix and suffix.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1375
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1369
  Illegal BamHI site found at 1351
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]