Part:BBa_K079050

GFP reporter protein under the control of the J23100 constitutive promoter and LexA 2 operator

This part is a DNA damage reporter. See experience part for more Experimental details.

The Northwestern 2019 iGEM team provided novel characterization data to part BBa_K079050. The 2008 Bologna Team reported their part was unable to be induced by UV light. However, after exposing it to various doses of UV-C (0 J/m^2, 50 J/m^2, and 125 J/m^2), we found that their part showed different responses for different amounts of UV exposure. Specifically, the rate of decrease in Fluorescence/OD is significantly different between each exposure time, providing evidence that this part may actually be able to differentiate between different UV doses, just not in the way it was originally designed for (Figures 3 and 4). Fluorescence measurements were taken every 3 minutes immediately after exposure in a 96-well plate reader for 3 hours.

Figure 3. Fluorescence/OD values for Part BBa_K079050 after exposure to various UV doses.

Figure 4. Fold Activation for part Part BBa_K079050 after exposure to various UV doses.
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
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 729
  

Contributions: Group: iGEM_TAU 2022 Summary: We have generated an optimized version of this construct, utilizing the ESO software (developed by TAU representative group for the iGEM 2020 competition), by the means of evolutionary stability of it: improving its ability to preserve functionality over long periods of time by avoiding loss-of-function mutations, taking into account different aspects, such as kinetic and thermodynamical parameters and Codon-biases. A link to our parts improvement page: https://parts.igem.org/Part:BBa_K4219000.