Part:BBa_K1065201:Design

Pxyl+GFP in an integrative Bacillus subtilis specific backbone

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal prefix found in sequence at 4773
  Illegal suffix found in sequence at 1
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 4773
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 4779
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 4773
  Illegal BamHI site found at 4757
  Illegal XhoI site found at 1011
  Illegal XhoI site found at 1929
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found in sequence at 4773
  Illegal suffix found in sequence at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found in sequence at 4773
  Illegal XbaI site found at 4788
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NgoMIV site found at 1893
  Illegal NgoMIV site found at 3032
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 2856
  Illegal BsaI.rc site found at 5467
  Illegal SapI.rc site found at 605
  

Design Notes

We assembled the construct through standard assembly starting from the Bacillus specific backbone BBa_K823024 and the GFP + strong RBS BBa_E0840.

Source

The backbone comes from the LMU Munich IGEM 2012 team, the GFPmutB was extracted in pSB1A2 from the IGEM 2012 repository.

References

1. [1]C.Derré I, Rapoport G, Msadek T,The CtsR regulator of stress response is active as a dimer and specifically degraded in vivo at 37 degrees, MOLECULAR MICROBIOLOGY, 2000 Oct;38 (2):335-47.


2. [2]Cormack, B.P., Valdivia, R.H., and S. Falkow. FACS-optimized mutants of green fluorescent protein (GFP). GENE 173: 33-38 (1996).