Composite

Part:BBa_K634007:Design

Designed by: Eric Walters   Group: iGEM11_Wisconsin-Madison   (2011-09-23)

Double selection cassette (tagRFP-sacB-kanR)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 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 648
    Illegal SapI.rc site found at 30


Design Notes

We are still in the process of optimizing this device, the following considerations are undergoing testing:

- tagRFP was taken from an expression vector, did it lose 5' components essential for high expression?

- Can we reduce the frequency of spontaneous loss of function mutations in sacB through protein fusion with kanR?

- Is the kanR RBS too strong for the desired resolution of susceptibility? Can a weakened RBS increase the range of the selection?

- Similarly, is the RBS of K322921 too weak? Are we not getting enough translation of sacB to kill the population?

Source

tagRFP came from Evrogen sacB and kanR came from the registry

References