Composite

Part:BBa_K2447017

Designed by: Chee Wai Kit (david)   Group: iGEM17_NUS_Singapore   (2017-10-17)


Constitutive E2 killing protein production

As part of the double plasmid system to activate the kill switch, this second plasmid (if this part is integrated into a suitable backbone like pE2K) will be constitutively producing the E2 Colicin protein, and actively killing the cells. Depending on the amount of IM2 proteins produced by the first plasmid (BBa_K2447016), the cells would still survive when the kill switch is not in the 'ON' state (low phosphate (close to 0 uM) and low temperature (below 36 degrees) condition).

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.

For more information, do visit our [http://2017.igem.org/Team:NUS_Singapore/Experiments experimental page] on wiki.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 89
  • 1000
    COMPATIBLE WITH RFC[1000]


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