Terminator

Part:BBa_K3956009:Design

Designed by: Chris Jung   Group: iGEM21_CCA_San_Diego   (2021-07-31)


PRM9 Terminator


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 52
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Choosing the right promoter-terminator pair was shown to increase expression widely based on a 2013 paper from KA Curran. They found that low-strength promoters with the highest strength terminators produced the highest expression.

According to the study “Use of Expression-enhancing terminators in Saccharomyces cerevisiae to increase MRNA half-life and improve gene expression control for metabolic engineering applications,” PRM9 yielded the highest fluorescence output of strains expressing yECitrine of the 34 terminators studied; additionally, it showed a significant growth rate increase in comparison to the others (Curran et al). Overall, a terminator’s potential for being one of high capacity was determined by both fluorescence expression levels and the ability to support higher expression - both criteria fulfilled by PRM9, with its growth rate and fluorescence output. For our project, in which we expressed collagen via yeast, we needed a strong promoter - thus necessitating the presence of a strong terminator.


Source

S. Cerevisiae Chromosome 1 186836...187732

References

SGD Project. https://www.yeastgenome.org/locus/S000000078. Accessed 6 September 2021.

Curran, K. A., Karim, A. S., Gupta, A., & Alper, H. S. (2013). Use of Expression-enhancing terminators in Saccharomyces cerevisiae to increase MRNA half-life and improve gene expression control for metabolic engineering applications. Metabolic Engineering, 19, 88–97. https://doi.org/10.1016/j.ymben.2013.07.001.