Part:BBa_K823021

pSB_Bs1C-lacZ (lacZ reporter vector for B. subtilis)

This part is a reporter vector for Bacillus subtilis. It integrates at the amyE locus. It has a ampicillin resistance for cloning in E.coli and chloramphenicol resistance for B.subtilis. The multiple cloning site is followed by a lacZ gene for reporter assays. This backbone is a BioBricked version of the B. subtilis vector pAC6.

This BioBrick is part of the LMU 2012 igem project [http://2012.igem.org/Team:LMU-Munich/Bacillus_BioBricks BacillusBioBrickBox]

Reference: [http://www.ncbi.nlm.nih.gov/pubmed/11902727 Stülke et al.]

The two constitutive promoters P_liaG and P_veg were evaluated with the reporter vector pSBBs1C-lacZ. Promoter activity leads to the expression of the β-galactosidase which directly correlates to the promoter activity. The β-galactosidase assay of the constitutive Bacillus promoters P_veg and P_liaG was repeated three times. Data show one representative result. In the beginning of the growth curve both promoters show only low activity. But then it increases to a maximum before it decreases to the begininng level after about seven hours (Data not shown). Summing up the course of activity of both promoters P_veg and P_liaG is very similar based on the growth curve. The highest β-galactosidase activity and therefore the highest activity of the promoter P_veg with a maximum of 65 Miller units can be found during the transition from the logarithmic to the stationary phase. This is about five times higher than the acitivity of the promoter P_liaG with a maximum activity of about 12 Miller Units.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 9771
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 9777
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 9771
  Illegal BglII site found at 8313
  Illegal BamHI site found at 22
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 9771
  Illegal suffix found at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 9771
  Plasmid lacks a suffix.
  Illegal XbaI site found at 9786
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NgoMIV site found at 4562
  Illegal NgoMIV site found at 6888
  Illegal NgoMIV site found at 8688
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI site found at 4738
  Illegal SapI.rc site found at 5820