Generator

Part:BBa_K4202007:Design

Designed by: LIAOLIAO GAO   Group: iGEM22_ZJU-China   (2022-09-25)


PHY-Oxylimited-SpaBCTS


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 179
    Illegal EcoRI site found at 1245
    Illegal EcoRI site found at 3776
    Illegal XbaI site found at 100
    Illegal XbaI site found at 2024
    Illegal SpeI site found at 95
    Illegal PstI site found at 2674
    Illegal PstI site found at 2992
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 179
    Illegal EcoRI site found at 1245
    Illegal EcoRI site found at 3776
    Illegal SpeI site found at 95
    Illegal PstI site found at 2674
    Illegal PstI site found at 2992
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 179
    Illegal EcoRI site found at 1245
    Illegal EcoRI site found at 3776
    Illegal BamHI site found at 2495
    Illegal BamHI site found at 4765
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 179
    Illegal EcoRI site found at 1245
    Illegal EcoRI site found at 3776
    Illegal XbaI site found at 100
    Illegal XbaI site found at 2024
    Illegal SpeI site found at 95
    Illegal PstI site found at 2674
    Illegal PstI site found at 2992
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 179
    Illegal EcoRI site found at 1245
    Illegal EcoRI site found at 3776
    Illegal XbaI site found at 100
    Illegal XbaI site found at 2024
    Illegal SpeI site found at 95
    Illegal PstI site found at 2674
    Illegal PstI site found at 2992
    Illegal AgeI site found at 2309
    Illegal AgeI site found at 3317
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 361
    Illegal SapI.rc site found at 2409
    Illegal SapI.rc site found at 4919
    Illegal SapI.rc site found at 6121


Design Notes

We directly clone the sequence from the gDNA of B.subtilis ATCC6633 strain, supposing the box structure would fit to the hypoxic promoter. However, it didn't work well.


Source

The oxygen limitation promotor and its RBS are learned from BBa_K302021. The subtilisin production part is cloned from B.subtilis ATCC6633 strain' gDNA. The basic vector is PHY shuttle plasmid.

References