Part:BBa_K3927003
EL222 Expression Driver
This composite part encodes for a 3C120-CYC-LacO promoter (BBa K3927002 ) controlling EL222 for a feed forward circuit, with Part BBa_K2407003 as a terminator
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 670
Illegal XbaI site found at 203 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 670
Illegal NheI site found at 185 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 670
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 670
Illegal XbaI site found at 203 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 670
Illegal XbaI site found at 203 - 1000COMPATIBLE WITH RFC[1000]
Description
The EL222 expression driver is a composite part consisting of parts BBa_K3927002(3C120-CYC-LacO), BBa_K3570021(NLS-VP16-EL222) and BBa_K2407003(CYCt). 3C120-CYC-LacO is activated by dimerization of NLS-VP16-EL222 in the presence of blue light, which further increases the amount of NLS-VP16-EL222 produced by this part. The amount of NLS-VP16-EL222 then increases in a positive feedback loop, activating both the 3C120-CYC-LacO within this part as well as any other parallel, C120 motif-containing promoters until blue light is removed and NLS-VP16-EL222 un-dimerizes.
Usage
EL222 expression driver can be used to increase and sustain expression of C120 dependent promoters for longer periods of time in S.cerevisiae. This also decouples NLS-VP16-EL222 expression from the cell’s native metabolism by providing a self-sustaining loop that doesn’t rely on native trans-activating elements.
Design
In order for EL222 expression driver’s feedforward loop to kickstart, NLS-VP16-EL222 has to be first expressed from a separate transcriptional unit to drive initial expression of this part. LacO element in 3C120-CYC-LacO also has to be de-repressed, thus LacI must either be absent or disabled by a secondary input, allowing for greater control over this increase in expression.
Characterization
| None |