General Cloning – Cupriavidus
The pMTL70000 plasmids are modular shuttle vectors for use in Cupriavidus necator (formerly Ralstonia eutropha) and other Gram-negative chemolithoautotrophs. They essentially comprise four modular parts that are combined through four, unique restriction enzymes with 8bp recognition sequences: AscI, FseI, PmeI and SfbI.
Parts comprise a Gram -ve replicon that functions in C. necator, a second Gram-ve replicon that supports plasmid maintenance in the E. coli shuttle host, a selectable marker, and an application specific module. Several of the replicons include origins of transfer that support mobilisation of the vector by conjugation. Parts are arranged relative to one another in the same order and orientation. Each exists as at least three variants – with a current total of 18 parts available1. These are distributed as six plasmids, which together contain all 18 modules and are available from the SBRC online shop.
- Muhammad Ehsaan, Jonathan Baker, Katalin Kovács, Naglis Malys, Nigel P. Minton. The pMTL70000 modular, plasmid vector series for strain engineering in Cupriavidus necator H16. Journal of Microbiological Methods, 2021, Oct;189:106323. https://doi.org/10.1016/j.mimet.2021.106323.