Figure 1.

Building blocks, DNA assembly, and replacement strategies for MultiSite Gateway cloning. A, Entry clones, available either as listed in Table I or potentially added to characterize sequences of interest. B, Assembly of expression clones by recombination of two (left) or three (right) entry clones with a destination vector in LR clonase reactions. C, Examples for the replacement of genetic elements by the counterselectable ccdB cassette within expression clones for the creation of modular destination vectors in reverse BP reactions. Box shape annotation is as indicated at the bottom of C. Green, blue, and red boxes mark sequences flanked by the att4-att1, att1-att2, and att2-att3 pairs, respectively. Gray boxes represent ccdB counterselectable cassettes. Gray lines indicate the backbone of binary T-DNA vectors.