Composite

Part:BBa_K634007:Design

Designed by: Eric Walters   Group: iGEM11_Wisconsin-Madison   (2011-09-23)
Revision as of 03:48, 27 September 2011 by Ewalters (Talk | contribs) (→‎Design Notes)

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?<p> Can we reduce the frequency of spontaneous loss of function mutations in sacB through protein fusion with kanR?<p> Is the kanR RBS too strong for the desired resolution of susceptibility? Can a weakened RBS increase the range of the selection?<p> Likewise, is the RFP 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===