Regulatory
NR1/2SV40

Part:BBa_K415509:Design

Designed by: Joy Jiao, Laura Deming, Adrian Slusarczyk   Group: iGEM10_MIT   (2010-10-27)

pNR1NR2_SV40 Shear Stress Responsive Promoter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 6
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 6
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 6
    Illegal BglII site found at 81
    Illegal XhoI site found at 77
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 6
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 6
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 42
    Illegal BsaI.rc site found at 62


Design Notes

MIT iGEM 2010 Mammalian Promoter Part.

Preliminary data suggests that this synthetic promoter lends transcriptional sensitivity to fluid shear stress in the HEK293FT cell line. The schematic for the construction of this part, based on Silberman et al., Angiogenesis, 2009, is shown below. The promoter consists of a concatenation of three different shear stress responsive motifs isolated from different genes in front of the SV40 minimal promoter.

 

Design of the NR1NR2_SV40 promoter

    Of these:

  • PDGF/B: the original shear stress responsive element (SSRE) isolated from the platelet derived growth factor B promoter and has sequence GAGACC.
  • Tissue Factor: Sp1 site isolated from the Tissue Factor promoter and has sequence GGGGCGGGGCG.
  • MCP-1: the TRE site isolated from the promoter of teh MCP-1 gene. It has the sequence TGACTCC.
  • SV40: Minimal promoter

Source

MIT iGEM 2010 Mammalian Part

References

Silberman et al. "Shear Stress-induced Transcriptional Regulation via Hybrid Promoters as a Potential Tool for Promoting Angiogenesis." Angiogenesis 12.3 (2009): 231-42. Print.