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Sadao Ota

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

Ag43 Display

 Source:  E. coli MG1655 genomic DNA

You are going to make an antigen 43 autotransporter display system based on this reference: PMID: 12107137. The gene encoding Ag43 is called flu, and the DNA sequence is here. You should also look at the protein sequence at YP_026164 which includes some annotation of the domains within the protein. The actual transporter portion of the protein is:

     Region          738..1017
                     /region_name="Autotransporter"

Upstream of this sequence is one of the passenger domains:

     Region          517..707
                     /region_name="PL2_Passenger_AT"

For your first part, you should make the part corresponding to amino acids 708-1017. For your second part, you should make the part corresponding to amino acids 517-1017.

Due to the long length of the second part, you will not sequence your plasmid completely. Just sequence the ends with ca998 and G00101 to confirm the first and last 500bp.

This part encodes a N-terminal display protein

Your displayer part should be of the {<part!} style (no start, with stop). There should be NO prepro sequence in your part. You should design your construction file to insert your part into plasmid pBca9495KC-Bca1144#5 using EcoRI and BamHI. The map of this plasmid is here.

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