Composite

Part:BBa_K2282013:Design

Designed by: Eliott LAFON   Group: iGEM17_IONIS-PARIS   (2017-10-02)


mRFP under heat inducible system pL/cI857


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 1745
    Illegal AgeI site found at 1857
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part sums up our final heat sensitive construction. The overall strategy we chose is complex and we were not sure it would work. Our bibliographic review and analysis of the Paris-Saclay iGEM 2017 project (using DNA thermometers) we decided to use strong promoters (Anderson BBa_J23100 with its respective RBS and pL promoter) to obtain higher expression levels. We had a slight problem in sequence design with IDT because their system detected our double use of double terminators as concatemers and did not let us introduce the same double terminator BBa_B0015 so we change the double terminator of the new part by an E.coli terminator operon.



Source

BBa_K22820013 part has been assembled to the pL promoter - RBS - Double terminator (E.coli his operon terminator). This double terminator was extracted from the strain E.coli K12 MG1655 complete genome, size length 4,641,652 pb in order to be different from the one in the part BBa_K22820012 and avoid concatemers problems.


References