Difference between revisions of "Part:BBa K1499252:Design"
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GFP from the registry was modified with the stop codons. | GFP from the registry was modified with the stop codons. | ||
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Submitted the designed construct for synthesis by Integrated DNA Technologies and cloned into pSB1C3. | Submitted the designed construct for synthesis by Integrated DNA Technologies and cloned into pSB1C3. | ||
===References=== | ===References=== | ||
Revision as of 22:23, 2 November 2014
GFP with 2 stop codons generator + 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]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 705
Design Notes
We included two amber stop codons to prevent the expression of GFP in organisms containing the amber release factors. The part could not be cloned in wild-type E. coli because the toxic supP tRNA acquired mutations or prevented transformation. So we had to use amberless E. coli to finally get the part cloned and sequenced without mutations.
Source
GFP from the registry was modified with the stop codons.
Submitted the designed construct for synthesis by Integrated DNA Technologies and cloned into pSB1C3.