Regulatory
M3 TTL

Part:BBa_K202003:Design

Designed by: Poonam Srivastava   Group: iGEM09_Minnesota   (2009-10-19)


Hybrid promoter having multiple operator sites.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 192
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

111 base pair oligos were designed for SOEing PCR with 18 base pair flanking region.


Source

The promoter was obtained by artificial synthesis. SOEing PCR was first done to combine two 111 base pair oligos, followed by the nested PCR. This strategy generated a 203 base pair hybrid promoter. This was directly cloned into pGLOW TOPO plasmid, upstream to the GFP cycle3. Biobrick suffix and prefix were later introduced through PCR and promoter fragment was cloned in plasmid pSBA1 upstream to the part E0240.

References