Part:BBa_K415510:Design
pSRE/CRE2_SV40 L4R1 Mammoblock
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 6
- 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 6
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 6
Illegal BglII site found at 82 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 6
- 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 6
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
We wanted to avoid cloning the entire FosB promoter; our goal here was to isolate the regions that specifically controlled mechanosensitivity. This SRE/CRE2 element had previously been shown in the literature to control the responsiveness of osteoblasts to shear stress, and to confer mechanosensitivity when concatenated in front of the SV40 promoter and transfected into mammalian cells. We used the shortest sequence found in the literature to still control the pressure-sensing response.
Source
We used primers containing the 37bp SRE/CRE2 element to add the sequence in front of an SV40 constitutive promoter via PCR. We cut the PCR product with PacI and BsrgI and ligated it into a cut L4R1 Mammoblock Promtoer Entry Vector.