Regulatory

Part:BBa_K1456015:Design

Designed by: Mustafa Yılmaz   Group: iGEM14_ATOMS-Turkiye   (2014-10-07)

NF-κB (Nuclear Factor kappa B) ROS response element

ATOMS KB results 1.png

Cloning

ATOMS KB results4.png

  • We cloned kB-RE and inserted it into the downstream region of CMV mini promoter as it’s shown above.
  • NF-kappaB (NF-kB) was synthesized to GenScript™ company and it came in pUC57 plasmid. We digested it with BamHI & PstI and exposed it to Antarctic phosphatase.

Afterwards, we purified our part via phenol chloroform method. We performed the same procedures onto the pTRE-luciferase vector. Eventually, we ligated them. We inserted our plasmid (pTRE-luciferase kB-RE) into DH5α strain and performed colony PCR by using CMV forward and kB reverse primers. At the end of this experiment, we expected a band seen in the 20-30 bp line.And we observed correct bands in the expected region.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1
    Illegal XhoI site found at 47
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Design Notes

none


Source

human

References