General Cloning

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The pMTL80000 plasmids are modular shuttle vectors for use in Clostridia and other anaerobes. 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 and –ve replicon, selectable marker, and application specific module. A fifth module comprising oriT/traJ may be added as a unique ApaI fragment should mobilisation by conjugative plasmid transfer be required. Parts are arranged relative to one another in the same order and orientation. Each exists as at least two variants – with a total of 18 parts, 395 different vectors were possible when originally published – https://doi.org/10.1016/j.mimet.2009.05.004. These are distributed as five plasmids, which together contain all 18 modules, and are available from the SBRC online shop.

A table showing vector nomenclature and available parts is shown below. Since publication, the number of modules has been expanded to include 5 new, Gram +ve replicons and a part encoding the FAST reporter protein. New modules are emboldened in the table below:

Example = pMTL83141

¹ = plasmid from Clostridium carboxidivorans P7
²MCS = Multiple Cloning sites
³FAST = Fluorescence-Activating and Absorption-Shifting Tag Protein