Difference between revisions of "Part:BBa K2669000:Design"
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2669000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2669000 SequenceAndFeatures</partinfo>
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<h2>Design Considerations</h2>
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<p>To make this part, we had to modify part BBa_K2003011 by moving the start codon to the front of the histidine tag, as opposed to after it.  We also wished to express this protein constitutively as opposed to inducibly in order to make it more convenient for us to test if our part worked.  In order to prevent potential protein aggregation and/or strain on the cells we expressed our part in a low copy ampicillin backbone ordered from IDT (pUCIDT).  In addition, VR and VF2 cloning sites were inserted into our part for convenience, but are not in this composite part.  </p>
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<p> We also further modified [[Part:BBa_K2003011]] by adding a stop codon (taa) at the end of the flexible GSG linker in order to use it in our composite [[Part:BBa_K2669000]].  It is <b>very important</b> to note that this stop codon should be removed/moved to the end of the coding sequence if one wishes to use this part to create a fusion protein expressing construct when using this composite part.  </p>
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<p>It should be noted that we did not test the flexible linker with another protein, as we did not design the flexible linker ourselves.  For more information, please visit the [[https://parts.igem.org/wiki/index.php?title=Part:BBa_K2003011]] 2016 Uppsala team page</p>

Revision as of 11:28, 30 September 2018

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Considerations

To make this part, we had to modify part BBa_K2003011 by moving the start codon to the front of the histidine tag, as opposed to after it. We also wished to express this protein constitutively as opposed to inducibly in order to make it more convenient for us to test if our part worked. In order to prevent potential protein aggregation and/or strain on the cells we expressed our part in a low copy ampicillin backbone ordered from IDT (pUCIDT). In addition, VR and VF2 cloning sites were inserted into our part for convenience, but are not in this composite part.

We also further modified Part:BBa_K2003011 by adding a stop codon (taa) at the end of the flexible GSG linker in order to use it in our composite Part:BBa_K2669000. It is very important to note that this stop codon should be removed/moved to the end of the coding sequence if one wishes to use this part to create a fusion protein expressing construct when using this composite part.


It should be noted that we did not test the flexible linker with another protein, as we did not design the flexible linker ourselves. For more information, please visit the [[1]] 2016 Uppsala team page