Part:BBa_K1725002
PhlF repressible promoter + weak 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 719
This reporter was used to characterise the promoter BBa_K1725000 which was used to drive expression of GFP with two different Ribosome Binding Sites, BBa_B0032 in E5501, for this part, and BBa_B0034 in I13500. PphlF is a stronger promoter than pL-tet (BBa_R0040 or PsrpR (BBa_K1725020). (figure 1) PhlF (BBa_K1725040) gave 83-fold repression of GFP expression from PphlF, whereas the control, TetR (BBa_C0040), gave only 33-fold repression of pL-tet. (figure 2)
Figure 1 Relative Promoter Strength. BioBricks: K1725082 (pL-tet driving GFP expression with B0034), E5504 (pL-tet driving GFP expression with B0032), K1725001 (PphlF driving GFP expression with B0034), and K1725002 (PphlF driving GFP expression with B0032). Mean and standard deviation of replicates were calculated to give value and error bars, and normalised against a negative control.
Figure 2 Fold Repression. BioBricks: K1725082 (pL-tet driving GFP expression), K1725031 (pL-lac driving TetR expression), K1725001 (PphlF driving GFP expression), and K1725042 (pL-lac driving PhlF expression). Repressor protein expression induced with 100μM IPTG. Mean and standard deviation of replicates were calculated to give value and error bars, and normalised against a negative control.
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