Difference between revisions of "Part:BBa K1499253:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | + | The aeBlue protein from Team Uppsala 2012, [https://parts.igem.org/Part:BBa_K864401 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 ([https://parts.igem.org/Part:BBa_K1499251 BBa_K1499251]) is used to translate the 3 UAG stops. | ||
===Source=== | ===Source=== |
Revision as of 16:47, 2 November 2014
aeBlue generator with 3 UAG stops + supP tRNA
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE 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
iGEM