Composite

Part:BBa_K3596049

Designed by: Xinyu Liu   Group: iGEM20_GreatBay_SZ   (2020-10-22)


BIOT RBS mut2


BIOT is a basic design for the expression of Geobacter sulfurreducens' pilin-based electrically conductive protein nanowires (e-PNs)[1]. The system includes a tac promoter which promotes the pilin gene(pilA) to express protein monomers; the monomers are later assembled into protein nanowires by the TypeIV pilin assembly system, which is constructed from the gene cluster at the downstream of the pilin gene(figure 1). The e-PNs can generate electric potential with the presence of water molecules in the air, which can be further modified for other purposes.

Figure 1: A Construction of BIOT gene circuit

In order to improve the protein nanowire, we mutated the RBSs of the TypeIV pilin assembly system. This part codes for BIOT with the ppdA RBS replaced by ppdA RBS mut2.The e-PN productivity of this part is shown in figure 2.

Figure 2: Different pilin productivity of engineered BIOT system with different RBS.

References

[1] Liu X, Gao H, Ward J E, et al. Power generation from ambient humidity using protein nanowires[J]. Nature, 2020, 578(7796): 550-554.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 2803
    Illegal PstI site found at 8393
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 8393
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2393
    Illegal BglII site found at 2857
    Illegal BglII site found at 4658
    Illegal BamHI site found at 1073
    Illegal BamHI site found at 3182
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 2803
    Illegal PstI site found at 8393
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 2803
    Illegal PstI site found at 8393
    Illegal NgoMIV site found at 1300
    Illegal AgeI site found at 1146
    Illegal AgeI site found at 1439
    Illegal AgeI site found at 2002
    Illegal AgeI site found at 4626
    Illegal AgeI site found at 6914
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 7920
    Illegal SapI.rc site found at 4262


[edit]
Categories
Parameters
None