Part:BBa_K526000
Pompc promoter coding fimE
fim inversion system
fimE is a invertase,by function,that is capable of inverting any DNA sequence that is flanked by fimE recognition site. When fimE recognition sequences are flanked to a promoter element we can ensure that there is no leaky expression as the promoter can be maintained in the opposite orientation till the presence of inducer. Thus, fim inversion promoters ensures that there is a tight regulation of target gene expression (Fig 1).
Light Receptor
Pompc is a promoter sequence that is regulated by the hybrid light receptor, Cph8. Unfortunately we can only inactivate a promoter with light. We need to have an invertor in order to activate gene expression with light. cI is being widely used as an inverter in light mediated gene expression
A Novel Invertor for Light Mediated Gene Expression
fim inversion promoter system is used to provide a tight control of gene expression. We made an attempt to introduce an one-way invertor to the light receptor using the fimE processor. In this biobrick part, we have made an attempt to regulate the expression of fimE based on light so that we could precisely turn on a previously uninduced gene. The system would be permanently on indicating need for no further inducer. This system would also ensure more tight control of target gene expression. This combination would be potentially useful for toxic gene expression as the basal level of the toxic gene would be kept as low as possible.
Contrary to the expectation, a combination of light inducible system and fim inversion system led to a constitutive gene expression (Fig 2). Hence, we hyphothesize that optimizing the RBS of fimE processor would help avoid this basal expression and favor a tight regulation of gene expression.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 750
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 750
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 750
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 750
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 750
- 1000COMPATIBLE WITH RFC[1000]
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