Reporter

Part:BBa_K2151007

Designed by: Matej Vucak, Katy Baker, James Provan   Group: iGEM16_Glasgow   (2016-08-30)


p25-I13500: GFP under control of medium constitutive promoter and strong ribosome binding site

This part consists of I13500 (GFP with a strong ribosome binding site) under the control of p25, a medium constitutive bacterial promoter. Its intended use is to serve as a promoter-reporter construct in lactic acid bacteria. p25 was initially identified in a promoter probe survey of the Streptococcus thermophilus genome, alongside weaker promoters p8 and p20.

We used p25 as a candidate promoter to drive gene expression in both Streptococcus thermophilus and E. coli. Quantification of p25 using GFP in E. coli revealed promoter strength on par with medium Anderson promoters (30.8% as strong as J23100, 43.7% as strong as J23101, 130% as strong as J23106).

T--Glasgow--B0034 Measurement.jpg

Figure 1: Mean fluorescence values for I13500 under the control of various promoters. Orange represents S. thermophilus promoters, blue represents Anderson family promoters. Error bars represent standard deviation for each set of samples.


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 707


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