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To perform in silico analysis, we used Clone Manager. All plasmids were first implemented into the software, and then used to help with cloning simulation, restriction, PCR etc.

Plasmid containing construct for chromosomal evolution

pTGD ccdA Pmb1 BCD7 GFP CmFRT

This plasmid contains the ccdA antitoxin and the reporter gene GFP. The homologous regions are necessary to perform Chemically Inducible Chromosomal evolution (CIChE).

Plasmids containing toxin ccdB

pSB3T5 T7 ccdB

pSB4A5 T7 ccdB

pSB6A1 T7 ccdB

New part

pSB1C3 T7 ccdB