Part:BBa_K3463008

J23100 RBS RhlR B0015

For our system to be as sensitive as possible to BHL, we needed to have the highest production of RhIR in the engineered E. coli strain. For that we tested three constitutive promoters with different strengths found in the iGEM Bank: J23100, J23102 and J23106. To identify the strongest one, we integrated the promoters into pUCBB-eGFP.

Then, we monitored the expression of the eGFP by measuring both the fluorescence and the absorbance of the bacterial cultures (transformed E. coli DH5 alpha). Thus we quantify the level of expression of eGFP as a function of time for the different constructs. After selection of the strongest promoter (J23100), we replaced the reporter gene by RhIR to build the pUCBB J13100-RhlR. To finish, we added the terminator B0015 behind RhlR .

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 275
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 750