Part:BBa_K1725001
PhlF repressible promoter + strong RBS + GFP
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 718
This reporter was used to characterise the promoter K1725000.
GFP fluorescence of K1725001, K1725002, K1725021, K1725022, K1725082, and E5504 with plasmid backbone pSB3K3 was measured to compare the relative strengths of promoters K1725000 and K1725020 to a promoter already well documented in the registry, R0040. The figure below shows the fluorescence scan image and a graph of approximate molecules of GFP per cell. These results indicated that K1725000 is a significantly stronger promoter than R0040 or K1725020.
All constructs with pSB3K3 plasmid backbone, in DH5α cells. Replicates of constructs and controls from three colonies, under the same conditions. Mean and standard deviation of replicates were calculated to give value and error bars.
This reporter was also used to characterise repression of K1725000 by K1725040.
K1725000 driven expression is repressed by K1725040 (phlF encoding PhlF repressor) as shown in the figure below. K1725042 is K1725040 driven by the lacI regulated promoter K1725080. Our control was K1725083 (the Tet repressor C0040 also driven by K1725080) and K1725082 (the TetR repressible promoter R0040 driving expression of I13500).
Represser constructs with pSB1C3 backbone; promoter driving GFP constructs with pSB3K3 backbone. Repressor protein expression induced with 100μM IPTG. Replicates of constructs and controls of three dilutions from one colony, under the same conditions. Mean and standard deviation of replicates were calculated to give value and error bars.
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