Change the plan to build upon Cambridge ideas

We were thinking to just slightly modify the plasmids that produce a gradient fluorescent band, in the article “A synthetic multicellular system for programmed pattern information” by Basu et al. A closer look at the plasmids sequence showed that our lab didn’t possessed the restriction enzymes nor adequate sticky ends in the sequence, required to cut and ligate biobricks insertion. Therefore we look after alternative solution for the project and found last years iGEM construct design very suitable for our bioclock construction. It would allow the team to modify their biobricks construct and rearrange to build band detect construct suitable for our bioclock project

Solve the detailed problems like AHL removal, color protein, and irreversible transmission of signal.

Overproduction of AHL was a problem that could destabilise the system. We thought the AHL gradient produced would disturb the quorum sensing signalling of the next set of band detect colonies in the bioclock. Therefore we planned to design a biobrick construct with the same organisation as the band detect construct, with the only difference that the fluorescent gene would be replaced by aiia-gene producing a protein degrading AHL.