Part:BBa_K1495000

KillerRed with Lactose Promoter

This part contains the gene called KillerRed. It has been codon optimized for use in E. coli. This functions as a safety piece that can be used for all project. The KillerRed gene causes the cells to produce reactive oxygen species within the cell when exposed to white light. A lactose promoter has been included as a proof of concept device. When provided with light and IPTG the cells would die, otherwise they would live.

Characterization by the 2015 CSU_Fort_Collins Team:

Construction: This part was transformed into two strains of E. coli, DH5alpha and NEB F'I^2. The DH5alpha strain was unable to fully repress the KillerRed even when there was no IPTG present. The F'I^2 cells did have that control. We sequenced verified in both strains.

Experimental Verification: We ran many experiments on both strains to determine the parameters necessary for this part to function. For detailed experimental plans see our wiki. We changed only small details from experiment to experiment, and could not find any significant difference between our control and our KillerRed cells. There was no apparent correlation between any of our data points. We have not included a figure as there is no logical way to present this data graphically. Our data tables are available below. We thus concluded that our KillerRed does not work the way we expected. We speculate that the E. coli may mutate the KillerRed gene so that the cells turn red but no reactive oxygen species are released to induce cell death.

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 AgeI site found at 374
  
• 1000
  COMPATIBLE WITH RFC[1000]