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John Yi Wang

Z domain
Source: Synthetic You will be making a basic part out of the Z domain of protein A. Protein A is interesting because it binds antibodies. Some quick info about Protein A is here: http://www-nmr.cabm.rutgers.edu/photogallery/proteins/htm/page16.htm. The earliest reference about Z domain is: PMID: 3507693

You should follow the paper trail about Z domain and find the most modern sequence that maximizes affinity to IgG's while minimizing size. Before you start writing up your construction file, you should show me (Chris) your reasons for picking the peptide that you have chosen. You will be making your part synthetically from oligos. If you can find a DNA sequence for the part, try to stay as close to that sequence as possible. If there is only protein sequence available, use GeneDesign to make a DNA sequence.

The synthetic gene you design may be short enough to do it by Overlap Extension. If not, you should use gene synthesis. We'll discuss this later in class, but let me know if you need to do this and I'll walk you through it.

FLAG Tag
Source: pBca1256-K112602   {} This part has already been constructed as a BglBricks basic part in plasmid pBca1256. You will be transferring the part using the Gateway reaction into the assembly vector. To do this, you will react the source plasmid with plasmid pBca1254AK. As background, you should read PMID: 12536251.



This part encodes a passenger protein
After performing the Gateway reaction, your product plasmid will be pBca9495AK-partname. To see what the part looks like in the product vector, look at [[Media:JCA_pBca9495AK-Bca1144.seq | this map]] of the RFP-expressing part Bca1144 in pBca9495AK.