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Fig. 2 | Microbial Cell Factories

Fig. 2

From: Segregationally stabilised plasmids improve production of commodity chemicals in glucose-limited continuous fermentation

Fig. 2

The layout of the genes and the Type IIS part ends used to make up plasmids pQR2401 and pQR2402, ends fully explained in Additional file 1. A The different parts used in the assembly aligned with their respective ‘part ends’ to create complementary overhangs for the correct assembly order. The backbone of pET29a was amplified with either ‘Part End 1’ (for pQR2401) or ‘Part End 3’ (for pQR2402) at one end and always ‘Part End 2’ at the other. B The resulting components used in a TypeIIS digestion/ligation one-pot assembly. The pentagon indicates the presence of T4 DNA ligase and the hexagon indicates the presence of the TypeIIS restriction Endonuclease BsaI. The ‘Parts’ described in (a), with the exception of the backbone, were all ligated into a storage vector prior to this step, as indicated by the solid line representing a circular DNA molecule. c The only complete plasmids that can arise from the described one-pot assembly. pQR2401 assembles with a promotor followed by the rfp and cimA3.7 genes and a terminator region (shortened to ‘term’). pQR2402 assembles with the same operon arrangement but with a cer multimer resolution region upstream

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