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Plasmids strategy

We are using TetR and LacI to inverse the repression, which allows positive selection. We will select at two levels: in vivo and in vitro, using either lysis cassette or RFP respectively:

• in vitro: We can test the binding of TetR directly on plates by measuring RFP fluorescence
• in vivo: The cells will grow on chemostat chips. If TetR binds to the recogintion sequence, the cells will lyse and we will be able to recover the DNA

We have one plasmid with the TetR gene, where we will introduce mutations and two plasmids for in vivo or in vitro selection: