Figure 2

<EGFP-V> Insertions in α _s EE . (A) Location of 35 <EGFP-V> insertions in the α_sEE coding region. The 13 labeled insertions marked above the coding region are in the correct reading frame and encode fluorescent fusion proteins (12 unique insertions, 1 duplication). The insertions are named for the 3 amino acids of α_s duplicated during transposition. The 22 unlabeled insertions (18 unique) marked below the coding region are those in which <EGFP-V> landed out-of-frame with respect to α_sEE. Redundant insertions are indicated by the number of clones recovered at that site (e.g. 2X, 4X). (B) Srf I digestion of the transposed clone, followed by religation, removes theKan^r selection cassette and produces the full-length fusion protein. In the final fusion protein, the EGFP domain is bordered by amino acid linkers encoded by the Tn5 MEs as well as the 9 bp duplication of the target sequence that is generated during the transposition process.