Regulatory

Part:BBa_K3384314

Designed by: Yiqing Zhang   Group: iGEM20_NJTech_China   (2020-10-26)


pprm1 Ultra

Multiple copies of putative Ste12 binding sites are prevalent in pheromone-responsive promoters of Saccharomyces cerevisiae, which are also known as pheromone response elements (PREs). pprm1 contains 3 copies of PRE which is in the opposite direction of the promoter. pprm1 Ultra is a modified pheromone-responsive promoter. It contains 6 copies of PRE in the opposite direction of the promoter.


Characterization

When assembled with promotor BBa_K3384314, GFP BBa_K3112009, and CYC1 terminator BBa_K3384311 in pRS415, this construct expressed GFP. Then the activity of these promoters can be quantitatively measured through flow cytometer. As is shown in figure 1, the fluorescence intensity of the pprm1 is higher than that of the pprm1 Ultra under high concentration treatment conditions of pheromones. The pheromone concentration has no significant effect on the GFP expression intensity under the control of pprm1 Ultra, indicating that this engineered pprm1 remains a stable expression level when induced by pheromone. As pprm1 Ultra can provide a stable expression level regardless of the fluctuations in external conditions, it is expected to be applied to gene expression regulation in cell factories.



NJTech China--pprm1 Ultra.png


Reference

1. LSengupta, P., and Cochran, B. H. (1990) The Pre and Pq Box Are Functionally Distinct Yeast Pheromone Response Elements, Molecular and Cellular Biology 10, 6809-6812.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
chassisSaccharomyces cerevisiae