Jlogan; Genome Engineering MOD 1

Name: Jennifer Logan Email: jlogan at mit dot edu

M13.1 Section Design
The modifications that were necessary for me as a Discoverer (I choose to focus on gene 3) were mainly to unstuff the overlapping regions of gene VIII and the promoter of gene III. I did this by first copying over the region of overlap between gene VIII and the promoter of gene III and then modified two bases within the gene VIII overlap sequence (the wobble positions)so that gene III can no longer exist within it. However, care was take that the amino acid sequence was maintained. Also, two restriction sites (ecoRI and XmaI) were added to create unique sticky ends. I then found another restriction site (NIaIII) and cut open inbetween gene III so that I could insert a myc epitope that I modified by adding sticky ends (but upon insertion would destroy the NIaIII site). As the table above indicates, I hoped to insert the myc epitope because since it is known as a good tag, when regulation of gene III does occur, the myc epitope will help us pinpoint more exactly when this occurs. I hope that this will help me discover more about the regulation of gene III. My Redesigned Phage Section