Composite

Part:BBa_K2688027:Design

Designed by: William Briand   Group: iGEM18_GO_Paris-Saclay   (2018-09-28)


LEE5_GFP_native_ΔTir


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 site found at 50
    Illegal BsaI.rc site found at 1003


Design Notes

In pSB1C3-LEE5*Δtir-gfp, the tir fragment was deleted so that the gfp start codon is at the position of the former tir start codon, which yielded BBa_K2688017. The introduced point mutation was then PCR corrected, yielding this biobrick.

Our results indicate that gfp expression is improved when this fragment is removed compared to the parental pSB1C3- LEE5*-gfp plasmid, and that H-NS protein is still able to control the remaining LEE5 region. Of note, Ler proteins are able to activate LEE5 promoter.



Source

LEE5 is a topic of research at the host lab, which provided us with the source plasmid pKK-LEE5-gfp.

References