Composite

Part:BBa_K1897016:Design

Designed by: Ang Shi Hui   Group: iGEM16_NUS_Singapore   (2016-10-10)
Revision as of 20:29, 15 October 2016 by Shihuiangle (Talk | contribs) (→‎Design Notes)

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LuxR + GFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 949
    Illegal NheI site found at 972
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 84
    Illegal AgeI site found at 992
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 740


Design Notes

There is one HA tag available for characterisation of the LuxR protein produced via western blot. Note that the stop codon for LuxR BBa_C0062 is shifted to after the HA tag. Also, the transcriptional terminators rrnBT1 (from BBa_B0010) + BBa_B0012 for LuxR was derived from BBa_B0015, with the first 8 base pairs removed.

Source

LuxR was obtained from biobrick part BBa_C0062, which was derived from Vibrio fischeri. The promoter was synthesized based on the sequence obtained from BBa_J23119. The ribosome binding site (RBS) was synthesized based on the sequence obtained from BBa_B0030. The transcription terminators were synthesized based on the sequence obtained from BBa_B0015.

GFP was obtained from biobrick part BBa_E0040, which was derived from Aequeora victoria. The promoter was synthesized based on the sequence obtained from BBa_R0062. The ribosome binding site (RBS) was synthesized based on the sequence obtained from BBa_B0030. The transcription terminators were synthesized based on the sequence obtained from BBa_B0015.

References

Mishin, A. S., Subach, F. V., Yampolsky, I. V., King, W., Lukyanov, K. A., & Verkhusha, V. V. (2008). The First Mutant of the Aequorea victoria Green Fluorescent Protein That Forms a Red Chromophore†. Biochemistry, 47(16), 4666-4673.