Coding

Part:BBa_K1033259:Design

Designed by: Anders Edlund   Group: iGEM13_Uppsala   (2013-08-22)
Revision as of 19:53, 3 October 2013 by AndersE (Talk | contribs)

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Toxin-antitoxin system from lactobacillus plantarum


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 71
    Illegal SpeI site found at 447
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 71
    Illegal SpeI site found at 447
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 71
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 71
    Illegal SpeI site found at 447
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 71
    Illegal SpeI site found at 447
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The system has been amplified without RBS and its natural promotor. Since that promotor has not been characterized as far as we know, several promoters will probably have to be tried to find one that provides stability without adding to much cost of expressing the toxin-antitoxin. There is also a version available with the natural promoter and RBS. The part still contains two illegal restriction sites for EcorI and SpeI. However since it doesn't matter where in your construct you put the system it can still be assembled with XbaI and PstI.


Source

The Lactobacillus plantarum NC7 plasmid p256.

References

Plasmid p256 from Lactobacillus plantarum represents a new type of replicon in lactic acid bacteria, and contains a toxin–antitoxin-like plasmid maintenance system, Microbiology, Elisabeth Sorvig et al. September 30 2004