Part:BBa_K2447016

Phosphate-dependent, temperature-sensitive cascaded system with IM2 expression

As part of the double plasmid system for our kill switch, Design of this cascade system is similar to our other part BBa_K2447015, with the only exception being the GFP coding sequence being replaced with E2 immunity protein. If this part is integrated into a backbone (e.g pE8K), the production of IM2 (anti-killing protein) would be controlled, depending on the temperature and phosphate stimuli.

With the addition of the second plasmid BBa_K2447017 that would be capable of constitutive production of E2 Colicin (an DNAse), this construct is useful to serve as a highly specific kill switch where under high phosphate concentration and/or when the temperature is higher than 36 degrees, there would be constant production of E2 immunity protein to sequester the killing protein E2. For the remaining scenario such as when the phosphate concentration is low and when temperature is lower than 36 degrees, the kill-switch would be 'activated' where the production of E2 immunity protein will be down-regulated causing accumulation of E2 Colicin and subsequent cell death.

Sequence and Features

Assembly Compatibility:

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

  Illegal NotI site found at 1026
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2085
  Illegal XhoI site found at 1311
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1614
  
• 1000
  COMPATIBLE WITH RFC[1000]