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John Yi Wang

Welcome to your project page!

For each part listed, you should:

  • Design oligos to make your part
  • Write up proper construction files and put it on the Construction Files page
  • Put your oligos on the Oligo Log page
  • Put your part sequence on the part document
  • Name your parts according to the list here

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.

This part encodes a passenger protein

Your passenger part should be of the {<part>} style (no start, no stop). There should be NO prepro sequence in your part. If your part is naturally secreted, you should encode only the active peptide. You should design your construction file to insert your part into plasmid pBca9495AK-Bca1144#5 using EcoRI and BamHI. The map of this plasmid is here.


FLAG Tag

 Source: pBca1256-K112602    {<FLAG>}

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 this map of the RFP-expressing part Bca1144 in pBca9495AK.

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