Generator

Part:BBa_K1499253:Design

Designed by: Raman Nelakanti   Group: iGEM14_StanfordBrownSpelman   (2014-10-08)
Revision as of 16:48, 2 November 2014 by Raman2004 (Talk | contribs) (→‎Source)


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

References