Part:BBa_K2500011:Design
AND Gate B: Synthetic Promoter Responsive to LldR and LuxR
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We reasoned that flanking the pLux promoter with O1 and O2 should result in an AND-gate behavior and relied on the previously characterized parts BBa_K1847007 (O1-pconst-O2), BBa_R0062 (plux) and BBa_C0062 (LuxR).
In design B of the AND gate, we adapted the architecture of BBa_K1847007. The Anderson promoter was exchanged with the pLux promoter such that a) the -35 and -10 regions of the new promoter were at the same position as that of the original one and that b) the total distance between O1 and O2 was kept the same as in the original part. Further, we duplicated the lldR binding sites O1 and O2 in order to achieve a molecular zipper mechanism which would lead to a tighter DNA-loop as more LldR repressors are bound. This design in turn requires even higher, tumor-specific concentrations of L-lactate to completely unzip the loop.
AND gate B was ordered as a gBlock with flanking restriction sites, allowing it to be combined with our downstream elements via restriction-ligation cloning.
Source
This part was synthetically designed and ordered as a g-block. This part was designed based on the previously characterized parts BBa_K1847007 (LldP/LldR, Plldr) and BBa_R0062 (Plux)/ BBa_C0062 (LuxR).