Difference between revisions of "Part:BBa K1652000:Design"

 
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<partinfo>BBa_K1652000 short</partinfo>
 
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===Design Notes===
 
===Design Notes===
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TSS in PGK1 is not well-defined, and there exists a region at the end of the promoter, about 100 bp long, that is very AT-rich. The promoter also contains two elements that resemble TATA boxes.
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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.
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pPGK1 does not have a well-defined transcription start site, and it possesses two sites resembling a TATA box (see the sequence box on the main page), 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.
  
  

Revision as of 21:12, 18 September 2015

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 on the main page), 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