Composite

Part:BBa_K4115043:Design

Designed by: Kaijun Wang   Group: iGEM22_ShanghaiTech_China   (2022-09-30)


lac UV5 promoter-nifA-PnifH-sfGFP-B0015


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 82
    Illegal PstI site found at 223
    Illegal PstI site found at 1087
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 223
    Illegal PstI site found at 1087
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 724
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 82
    Illegal PstI site found at 223
    Illegal PstI site found at 1087
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 82
    Illegal PstI site found at 223
    Illegal PstI site found at 1087
    Illegal NgoMIV site found at 1344
    Illegal NgoMIV site found at 1539
    Illegal NgoMIV site found at 1614
    Illegal AgeI site found at 973
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1182
    Illegal BsaI.rc site found at 2101
    Illegal SapI site found at 405
    Illegal SapI site found at 1026


Design Notes

According to some papers [1], we speculate that IPTG induction experiments may be more successful in an anaerobic environment or in a medium with low nitrogen and no nitrogen.


Source

The sequence of the PhifH gene and nifA gene are gained from the genome of A.caulinodans ORS571 strain. The fragments are synthesized by the company.

References

[1] Ryu, MH., Zhang, J., Toth, T. et al. Control of nitrogen fixation in bacteria that associate with cereals. Nat Microbiol 5, 314–330 (2020). https://doi.org/10.1038/s41564-019-0631-2