FIG 6.

Golden Gate cloning vectors to construct tdFAST2-tagged proteins. The plasmids pMaFAST(C) and pMaFAST(N) encode the tdFAST2 gene which is codon-optimized for expression in Methanosarcina. Golden Gate cloning (BsaI) can be used to replace the lacZ cassette with a protein-encoding sequence of interest leading to a C- or N-terminal fusion to the tdFAST2. Nucleotide sequence of the cloning sites are shown below. The BsaI sites, ribosome-binding site (rbs) and the tdFAST2 open-reading frame is indicated.