An improved aminoglycoside resistance gene cassette for use in gram-negative bacteria and Streptomyces

A cloning cassette, carrying a modified amino-glycoside resistance gene (neo) from transposon Tn5 was constructed. Three restriction sites internal to the neo gene were eliminated by in vitro mutagenesis, allowing their use in designing new cloning vectors. The original, suboptimal transcription promoter was replaced with a synthetic sequence corresponding to the consensus for E. coli and Streptomyces promoters. The cassette has numerous restriction sites for easy subcloning of the promoter, the coding sequence or the whole gene.