Composite
Part:BBa_K2688027:Design
Designed by: William Briand Group: iGEM18_GO_Paris-Saclay (2018-09-28)
LEE5_GFP_native_ΔTir
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE 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.