A multipurpose Tn7-based transposon for mutagenesis of yeast DNA. (A) Schematic diagram of the multipurpose mini-Tn7 transposon indicating left and right termini, bacterial and yeast selectable markers, lox sites, and triplicate HA epitope-coding sequence. The 5′-truncated lacZ reporter and selectable markers are indicated by arrows. The epitope insertion element (or HAT tag) is indicated below the brackets depicting Cre-lox recombination. (B) DNA sequence of the Tn7 truncated left (Tn7L) and right (Tn7R) termini. (C) DNA and amino acid sequence of the Tn7-encoded HAT tag. Tn7 transposition results in a 5-base pair duplication of target site sequence. As the exact sequence of this duplication depends upon the site of Tn7 insertion, the duplicated residues are indicated here as five unspecified (N) carboxy-terminal nucleotides.
