Part:BBa_K3886015

RBS-Kid Toxin

This is a translation unit that can express Kid toxin protein in E. coli. Users can easily install a promoter like the L-arabinose pBAD promoter to initiate the transcription of it.

Characterization

In the Hidro system, this composite part is part of a genetic kill switch.

The kill switch contains a user-customizable inducible promoter and a toxin gene. In this project, we chose L-arabinose-inducible promoter and Kid toxin as an example. The kid is a toxin protein of the post-segregational killing system and can work as an endonuclease enzyme to cleave mRNA. The kill switch was integrated into the E. coli Top10 genome using the CRISPR-Cas9 system, thus maintaining the stability of the suicide genetic circuit.

After tracking the escaped bacteria, we can activate the kill switch in the engineered strain, thus eliminating any potential risk of escape. Kid kill switch has an escape efficiency lower 10^-3, according to the data shown in Figure 1.

Figure 1: The viability of E. coli with the Kid kill switch with or withour L-arabinose inducer

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 206
  
• 1000
  COMPATIBLE WITH RFC[1000]