Composite

Part:BBa_K1499252:Design

Designed by: Raman Nelakanti   Group: iGEM14_StanfordBrownSpelman   (2014-08-27)
Revision as of 22:23, 2 November 2014 by Raman2004 (Talk | contribs) (→‎Source)


GFP with 2 stop codons generator + 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
    INCOMPATIBLE 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.

References