Coding

Part:BBa_K4825024:Design

Designed by: Cai Luxi   Group: iGEM23_GreatBay-SCIE   (2023-10-11)


pXyl


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 1
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 1
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 1
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 1
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

"1. T20N2 spacer before the first xylose operator and T24N11 spacer between the two xylose operators contain mutations to suppress recombination;

2. Transcription start site (TSS) is connected with a 10bp Kozak sequence to increase the efficiency of translation;

3. The TATA box is placed 4bp down away from T20N2 spacer to result in a strong promoter that is independent on the carbon sources for cellular growth.

4. Red fluorescence protein (RFP) is attached after the promoter to test the functionality of the promoter. "


Source

Synthesized

References

"Chen, Y., Zhang, S., Young, E.M. et al. Genetic circuit design automation for yeast. Nat Microbiol 5, 1349–1360 (2020). https://doi.org/10.1038/s41564-020-0757-2.

Wei W, Shang Y, Zhang P, Liu Y, You D, Yin B, Ye B. Engineering Prokaryotic Transcriptional Activator XylR as a Xylose-Inducible Biosensor for Transcription Activation in Yeast. ACS Synth Biol. 2020 May 15;9(5):1022-1029. doi: 10.1021/acssynbio.0c00122. Epub 2020 Apr 17. PMID: 32268060."