BioDesign week 3 homework

We were given the assignment to effect changes in the GFP protein by changing specific residues using PCR and Golden Gate Assembly into pUC19. The resulting engineered proteins should be blue and cyan.

We were given a list of 14 oligos to buy, and the enzyme Bbs1-HF.

One pair of oligos were primers that annealed to regions of pUC19 will allow us to get a large part of the plasmid to work with, including the Amp-R and ori regions. Then there was a set of primers that targeted the first half of GFP, and a set of overlapping primers that targeted the c-terminal end. This latter set had tails that accomplished single AA changes that will hopefully result in emission wavelength changes.