Team:British Columbia/pBAD

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Overview

In order to make an analog biosensor, we need our traffic light to be able to respond differently to different concentrations of an input. However, the Registry is lacking in variable strength inducible promoters. We designed two variants of the pBAD promoter, one weaker and one stronger, based on AraC binding experiments performed by Niland et al.

pbad wt      ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCATTTTTATCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...
pbad weak    ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCTTTTTTATCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...
pbad strong  ACATTGATTATTTGCACGGCGTCACACTTTGCTATGCCATAGCAAGATAGTCCATAAGATTAGCGGATCCTACCTGACGCTTTTT...

Above is a sequence alignment of our promoter variants with the wild type pBAD, truncated for space. For the full sequences, see our parts in the registry at (strong) and (weak).

Here is the experimental data regarding the strength of these promoters when coupled to RFP: pBAD_RFP_Timecourse_3.xls Timecourse.jpg