Expanding the SiMPl Plasmid Toolbox for Use with Spectinomycin/Streptomycin

We recently developed the SiMPl plasmid toolbox, which is constituted by pairs of plasmids, generically indicated as pSiMPlx_N and pSiMPlx_C, can be stably maintained in Escherichia coli with a single antibiotic x. The method exploits split intein gp41–1 to reconstitute enzyme conferring resistance toward x, whereby each fragment expressed from one plasmids pair. pSiMPl are currently available for use ampicillin, kanamycin, chloramphenicol, hygromycin, puromycin. Here, we introduce another pair spectinomycin/streptomycin, broadening application spectrum toolbox. To find functional splice sites aminoglycoside adenylyltransferase, apply streamlined strategy looking exclusively at flexibility native cysteine serine residues, first validated splitting enzymes hygromycin. This could used future other antibiotics.