Difference between revisions of "Part:BBa K2587000:Design"
 
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<h2><b>Experimental design</b></h2>
 
<h2><b>Experimental design</b></h2>
  
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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 <i>S.cerervisiae</i> 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 <i>S.cerevisiae</i>.  
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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 optimized this sequence for the organism <i>S.cerervisiae</i> 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 optimizing a gene, usually not used for <i>S.cerevisiae</i>.  
  
 
__NOTOC__
 
__NOTOC__
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===Source===
 
===Source===
  
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The source of this part is the organism <i>Alivibrio fischeri</i> which was retrieved from Genbank (and is codon optimised for <i>S. cerevisiae</i>.)
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The source of this part is the organism <i>Alivibrio fischeri</i> which was retrieved from Genbank (and is codon optimized for <i>S. cerevisiae</i>.)
  
 
===References===
 
===References===
 
(1) <b>A Highly-characterized Yeast Toolkit for Modular, Multi-part Assembly.</b> Lee ME, DeLoache, WC A, Cervantes B, Dueber, JE. <i>ACS Synthetic Biology</i> 2015. DOI: 10.1021/sb500366v.
 
(1) <b>A Highly-characterized Yeast Toolkit for Modular, Multi-part Assembly.</b> Lee ME, DeLoache, WC A, Cervantes B, Dueber, JE. <i>ACS Synthetic Biology</i> 2015. DOI: 10.1021/sb500366v.

Latest revision as of 16:45, 17 October 2018

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 optimized 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 optimizing 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 optimized 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:
T--Duesseldorf--syntheticpromoter.PNG


Source

The source of this part is the organism Alivibrio fischeri which was retrieved from Genbank (and is codon optimized 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.