Difference between revisions of "Part:BBa K1499253:Design"

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===Source===
 
===Source===
  
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aeBlue - Team Uppsala 2012 iGEM
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aeBlue - Team Uppsala 2012 iGEM.
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Synthesized using Integrated DNA Technologies
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 +
Synthesized using Integrated DNA Technologies and cloned into pSB1C3
  
 
===References===
 
===References===

Latest revision as of 22:20, 2 November 2014


aeBlue generator with 3 UAG stops + supP tRNA


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 Notes

The aeBlue protein from Team Uppsala 2012, BBa_K864401, was codon optimized differently and the ATG start codon was removed. This was to introduce an N-terminal FLAG-tag to detect the different size products that could result from the amber stop codons we introduced in the protein.

We substituted 3 leucine codons with TAG, which allows us to test the functioning of the protein in amberless cells and prove the orthogonality of this system. We also included a C-terminal His-tag to be able to purify the complete protein product.

Finally, the supP tRNA (BBa_K1499251) is used to translate the 3 UAG stops.

Source

aeBlue - Team Uppsala 2012 iGEM.

Synthesized using Integrated DNA Technologies and cloned into pSB1C3

References