Composite

Part:BBa_K3739063

Designed by: Shichen Geng   Group: iGEM21_XMU-China   (2021-09-10)


pDawn-B0030-blrA-rrnB T1

pDawn promoter system can be activated by blue light. blrA can be activated by blue light and produce reactive oxygen species(ROS).


Biology and Usage

pDawn is a kind of regulation system that has been illustrated effectively in Vibrio natriegens which based on optogenetics. Under the irradiation of blue light, a series of regulatory elements will work on regulating the expression of genes that inserted into multiple cloning site In its downstream.

BlrA, originating from Bacillus subtilis, is a blue light-induced GTP-binding receptor, which possesses the LOV domain and autofluorescence. LOV domain enables it to be activated by blue light and produce ROS, which can damage the bacterial structure.

We take blrA as the core of our kill switch, combine it with inducible promotor system, terminator and a series of necessary elements. The whole part is inserted into a plasmid backbone and finally transformed into E.coli to gain enough correct and functional circuits. The circuits are transformed into Vibrio.natriegens. We choose growth curve and CFU to measure the killing effect of BlrA.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2171
    Illegal XhoI site found at 3851
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 63
    Illegal NgoMIV site found at 195
    Illegal NgoMIV site found at 289
    Illegal NgoMIV site found at 582
    Illegal NgoMIV site found at 1076
    Illegal NgoMIV site found at 1094
    Illegal NgoMIV site found at 1184
    Illegal AgeI site found at 414
    Illegal AgeI site found at 1542
    Illegal AgeI site found at 3703
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1643
    Illegal BsaI.rc site found at 525
    Illegal BsaI.rc site found at 3602
    Illegal SapI site found at 3920


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