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Nikhil Sharma

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

TraT

 Source:  pOX38

You will be making a C-terminal display system out of the TraT protein from F plasmid. pOX38 is just a particular variant of the F plasmid. You should read PMID: 9933744 and PMID: 10725535 for background. Unlike in these reference, you will remove the prepro region of the protein in your part. You should make the same C-terminal sequence present in the references as the last amino acids before the BamHI site. You will not be including any of the thrombin site/other modifications that were done in these references.

The sequence of TraT is available at NC_002483 in pubmed.

You can use the online tool at http://phobius.cbr.su.se/ to locate the prepro sequence.

This part encodes a C-terminal display protein

Your displayer part should be of the {<part>} style (no start, no stop). The source sequence for your part likely contains a prepro sequence. You should omit that region of the sequence in designing your part.

You should design your construction file to insert your part into plasmid pBca9495CA-Bca1144#5 using EcoRI and BamHI. The map of this plasmid is here.


Polystyrene-Binding Peptide

 Source:  Synthetic

For background, read PMID: 19056299. Make a BglBrick part encoding the peptide:

 FKFWLYEHVIRG

no start, no stop, in frame.

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.

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