(Solved): You now plan to clone the PCR amplicon for the WT FANCG gene into the PvuII site of plasm pGree ...

You now plan to clone the PCR amplicon for the WT FANCG gene into the PvuII site of plasm pGreen as illustrated below. The PCR product is a linear, blunt-ended dsDNA molecule of 36 bp, with an AfaI cleavage site at position 354. pGreen is 2000 bp is size and contains the GF (green fluorescent protein) gene, which upon expression in a bacterial hosts results in gree colonies. It also contains a penicillin resistance gene $\left({\mathrm{Pen}}^R\right)$ and three unique RE sites with CL site positions and distances between them as indicated on the diagram. a) Why is PvuII the only suitable RE site on pGreen for cloning of the insert? [1] b) You prepare a linear pGreen plasmid by PvuII digestion, ligate it to the PCR product, and transform it into bacteria. What phenotype would you expect for bacteria that have taken up recombinant plasmids, versus those with non-recombinant plasmids? [2]