Coding

Part:BBa_K3989024

Designed by: Tianyu Xu   Group: iGEM21_UZurich   (2021-10-01)


EsaR-PesaRC-GFP Testing construct of the promoter PesaRC( BBa_K3989008 ).

Structure

The goal is to test the sensitivity of PesaRC to a certain AHL molecule: 3OC6HSL. See the plasmid map in figure 1.

21 UZurich EsaR PesaR GFP.jpeg

Figure 1.The structure of the promoter testing construct. This construct consists three parts: a constitutively expressed EsaR protein controlled by lac operon, the PesaRC promoter and a reporter GFP gene downstream. The EsaR protein can be changed by its variants( BBa_K3989003 , BBa_K3989004 , BBa_K3989005 ) and so does the promoter( BBa_K23989009 ). If one wants to use the promoter later after the test, the GFP can also be changed to other functional gene.


Function

The way it works is rather simple: first we make sure the EsaR protein's existence. After the expression of the protein, it can bind to the promoter downstream and regulate the GFP expression under the control of the AHL molecule. In this case, the existence of AHL molecule will up-regulate the expression of the GFP. Thus, we can see a green fluorescence if we culture the cell containing this construct with AHL molecule. A similar construct is designed to test the promoter PesaS( BBa_K3989009 ).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 973
    Illegal BamHI site found at 1336
    Illegal BamHI site found at 1364
    Illegal XhoI site found at 1056
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2097
    Illegal BsaI.rc site found at 1371
    Illegal BsaI.rc site found at 2020


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