Translational_Unit

Part:BBa_K216006:Design

Designed by: Edinburgh iGEM 2009   Group: iGEM09_Edinburgh   (2009-10-16)

pentaerythritol tetranitrate reductase with native rbs


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 321
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 783


Design Notes

No special considerations, but due to an error, the primers were actually designed based on the homologous gene of another strain of Enterobacter cloacae, which has a very similar coding sequence but different sequence in the upstream region. The sequence shown here is the correct sequence determined by sequencing the construct.


Source

This part was amplified from plasmid pONR1 which bears the PETN reductase gene originally cloned from Enterobacter cloacae PB2.

References

Binks, P.R., French, C.E., Nicklin, S., & Bruce, N.C. 1996. Degradation of pentaerythritol tetranitrate by Enterobacter cloacae PB2. Applied and Environmental Microbiology 62(4), 1214-1219.

French,C.E., Nicklin,S. and Bruce,N.C. 1996. Sequence and properties of pentaerythritol tetranitrate reductase from Enterobacter cloacae PB2. J. Bacteriol. 178 (22), 6623-6627.

French, C.E., Nicklin, S., and Bruce, N.C. 1998. Aerobic degradation of 2,4,6-trinitrotoluene by Enterobacter cloacae PB2 and by pentaerythritol tetranitrate reductase. Applied and Environmental Microbiology 64, 2864-2868.

French, C.E., Rosser, S.J., Davies, G.J., Nicklin, S., and Bruce, N.C. 1999. Biodegradation of explosives by transgenic plants expressing pentaerythritol tetranitrate reductase. Nature Biotechnology 17, 491-494.