Composite

Part:BBa_K2207017:Design

Designed by: Junming Qian   Group: iGEM17_ZJU-China   (2017-10-26)


phlCD enzyme


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 3244
    Illegal XbaI site found at 4654
    Illegal SpeI site found at 1798
    Illegal SpeI site found at 5081
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 1798
    Illegal SpeI site found at 5081
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1399
    Illegal XhoI site found at 4737
    Illegal XhoI site found at 4904
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 3244
    Illegal XbaI site found at 4654
    Illegal SpeI site found at 1798
    Illegal SpeI site found at 5081
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 3244
    Illegal XbaI site found at 4654
    Illegal SpeI site found at 1798
    Illegal SpeI site found at 5081
    Illegal NgoMIV site found at 2309
    Illegal AgeI site found at 1090
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 3941


Design Notes

We constructed a expression system according to the mechanism of M2S intergration technique, so that we can intergrate multiple genes into the chromosome.


Source

We successfully cloned the phlCD via colony PCR from Pseudomonas fluorescens 2P24, which is a gift from Prof. Liqun Zhang of China Agriculture University.

References

Li S, Ding W, Zhang X, et al. Development of a modularized two-step (M2S) chromosome integration technique for integration of multiple transcription units in Saccharomyces cerevisiae[J]. J Immunoassay Immunochem, 2016, 9(1):232.