Reporter

Part:BBa_K2151003

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


pHLBA-E5501: GFP under control of strong constitutive promoter

This part consists of E5501 (GFP with medium ribosome binding site) under the control of pHLBA, a strong constitutive bacerial promoter. Its intended use is to serve a promoter-reporter construct in lactic acid bacteria. pHLBA was initially identified in studies of Lactobacillus bulgaricus where it drives the expression of histone-like proteins. pHLBA is likely functional in all lactic acid bacteria.

We used pHLBA as a candidate promoter to drive gene expression in both Streptococcus thermophilus and E. coli. Quantification of pHLBA using GFP in E. coli revealed promoter strength on par with strong Anderson promoters (93% as strong as J23100, 132% as strong as J23101).

T--Glasgow--B0032 Measurement.jpg

Figure 2: Mean fluorescence values for E5501 under the control of various promoters. Red represents S. thermophilus promoters, blue represents Anderson family promoters. Error bars represent standard deviation for each set of samples. Promoters are ordered according to their strength when expressing I13500.


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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