Geobacillus thermoglucosidasius is a
gram-positive thermophile of industrial interest that
exhibits rapid growth and can utilize a variety of
plant-derived feedstocks. It is an attractive chassis
organism for high temperature biotechnology and
synthetic biology applications but is currently limited
by a lack of available genetic tools. Here we describe
a set of modular shuttle vectors, including a
promoter library and reporter proteins. The compact
plasmids are composed of interchangeable modules
for molecular cloning in Escherichia coli and stable
propagation in G. thermoglucosidasius and other
Geobacillus species. Modules include two origins of
replication, two selectable markers and three reporter
proteins for characterization of gene expression.
For fine-tuning heterologous expression from these
plasmids, we include a characterized promoter library
and test ribosome binding site design. Together,
these gene expression tools and a standardized
plasmid set can facilitate modularity and part exchange
to make Geobacillus a thermophile chassis for
synthetic biology.