Part:BBa_K1088008

B. subtilis dxs-GFP fusion (lac promoter without lac inhibitor: IPTG uninducible)

This part consist of the dxs gene derived from B. subtilis fused to GFP at the translational level with a 10 AA linker between the proteins. The reporter fusion is under the control of the lac promoter and has a strong RBS.

The purpose of the part was to test the expression profile of dxs from the lac promter. BBa_K1088011 is a similar part that does not contain the linker-GFP part.

Fluorescence activated cell sorting (FACS) was used to examine the expression profile the reporter fusion. The part was examined in the K-12 MG1655 (natural levels of LacI) and KG22 (overexpression of LacI from chromosome) strains. The experiment proved that overexpression of LacI is required for repression of the lac promoter (and therefore also control of the regulation). BBa_K1088026 is a similar part which overexpresses LacI from the part itself.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1279
  Illegal EcoRI site found at 1936
  Illegal PstI site found at 1337
  Illegal PstI site found at 1783
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1279
  Illegal EcoRI site found at 1936
  Illegal PstI site found at 1337
  Illegal PstI site found at 1783
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1279
  Illegal EcoRI site found at 1936
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1279
  Illegal EcoRI site found at 1936
  Illegal PstI site found at 1337
  Illegal PstI site found at 1783
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1279
  Illegal EcoRI site found at 1936
  Illegal PstI site found at 1337
  Illegal PstI site found at 1783
  Illegal NgoMIV site found at 1236
  Illegal AgeI site found at 1129
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1078
  Illegal BsaI.rc site found at 2792
  Illegal SapI.rc site found at 1777