Regulatory

Part:BBa_K1652000:Design

Designed by: Martin Hanzel   Group: iGEM15_uOttawa   (2015-09-08)
Revision as of 21:13, 18 September 2015 by Arthanzel (Talk | contribs) (→‎Design Notes)

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pPGK1Gx


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]


Design Notes

This promoter, pPGK1Gx, is based on the PGK1 (phosphoglycerate kinase) promoter from S. cerevisiae. As a promoter driving the production of a glycolytic enzyme, this promoter has extremely high, constitutive expression. Two Gal4 binding sites have been added downstream of a TATA-like element so that this promoter may be repressed with the Gal4 protein or its derivatives (like GEV). The close proximity of the binding sites to the transcription start site creates steric hindrance and prevents RNA polymerase from binding, thus repressing the promoter.

pPGK1 does not have a well-defined transcription start site, and it possesses two sites resembling a TATA box (see the sequence box above), so it is not clear where the optimal location is for placing a binding site that represses the promoter as much as possible. In addition, the last ~100 bp of the promoter are extremely AT-rich, so it is difficult to estimate the location of the TSS and polymerase binding site.

Source

S. cerevisiae

References