Composite
occupyDNAP

Part:BBa_M36549:Design

Designed by: Rashmi Sharma, Bryce Bajar, Kendrick Wang   Group: Stanford BIOE44 - S11   (2011-12-08)

PoPS --> N4 gp8 (DNA replication inhibitor)


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This composite part can successfully be used in K12 E. coli on a vector with ampicillin resistance and a rhamnose-inducible and glucose-repressible sensor (see Experience). This sensor contained rhaR, rhaS, and promoter rhaB (see reference 2 below for more imformation), and was physically directly before the RBS of the construct. It was found that this construct is difficult to synthesize because of slight leaky expression, but it was successfully synthesized in two tries. One could consider, if coupling this construct with a sensor, to ask the synthesis company to include the repressor of the sensor while cloning (glucose in this case) to minimize failure.

Source

[http://www.parts.igem.org Registry of Standard Biological Parts]
[http://biofab.jbei.org/services/studio/dac/ BIOFAB Data Access Client]

References

Yano, S.T. and L.B. Rothman-Denes (2011), A phage-encoded inhibitor of Escherichia coli DNA replication targets the DNA polymerase clamp loader. Molecular Microbiology, 79: 1325-1338.
Giacalone, M.J., Gentile, A.M., Lovitt, B.T., Berkley, N.L., Gunderson, C.W. and M.W. Surber. (2006), Toxic protein expression in Escherichia coli using a rhamnose-based tightly regulated and tunable promoter system. BioTechniques, 40: 355-364.