Figure 3: The Tol2 Multisite Gateway Cloning strategy: Att site flanked promoter and cDNA (gene of interest) fragments are generated by PCR and combined with pDONRP4-P1R and pDONRP221 vectors in the presence of BP Clonase to yield 5’ and middle entry clones, respectively. The two entry clones and a readily available 3’ entry clone (p3E-polyA) are combined together along with a destination vector pDestTol2CG2 in the presence of LR Clonase to yield an expression clone.