Coding

Part:BBa_K2587000:Design

Designed by: Ylenia Longo   Group: iGEM18_Duesseldorf   (2018-09-15)
Revision as of 15:39, 19 September 2018 by Yllon (Talk | contribs)

Experimental design

Part of our project was to design a synthetic promoter, able to induce expression of a reporter gene, which is activated upon synthesis of the Quorum sensing molecule acyl homoserine lactone synthase, by LuxI. Therefore we codon optimised this sequence for the organism S.cerervisiae and implemented this in our construct. With this we did not only apply this sequence into our project, but also improved a previous part by codon optimising a gene, usually not used for S.cerevisiae.


luxI_codon optimized S.cerevisiae


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 6
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 689
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 30
    Illegal BsaI.rc site found at 673


Design Notes

This part is codon optimised for S.cerevisiae and TypeII S restriction sites (on both sides) are added to the sequence for special use in the YTK toolbox but also for general Golden Gate cloning, a trending becoming cloning method. This construct was built using the MoClo YTK Toolbox (1). As an example, our final construct containing the LuxI gene is shown here:
<img src="T--Duesseldorf--syntheticpromoter.PNG">


Source

The source of this part is the organism Alivibrio fischeri which was retrieved from Genbank ( and is codon optimised for S.cerevisiae.

References

(1)A Highly-characterized Yeast Toolkit for Modular, Multi-part Assembly. Lee ME, DeLoache, WC A, Cervantes B, Dueber, JE. ACS Synthetic Biology 2015. DOI: 10.1021/sb500366v.