Part:BBa_K2244007

ColE promoter +mhei gene +mcherry gene +T1terminator

The device is a functional plasmid containing a MBC hydrolase gene (mheI) (BBa_K2244004), which encodes carbendazim hydrolase. In order to study expression of target gene in cytoplasm, mheI was fused to a reported gene mCherry (BBa_J06504), which functions as indicator of the uptake of mheI. This part is regulated under the control of ColE promoter, which is from E. coli ColE1 plasmid.

Biology

ColE promoter(BBa_K2244006) sequence is derived from the promoter region of colicin E gene located in the ColE1 plasmid of E.coli. ColE promoter contains a ‘SOS’ operator region that allows the binding of LexA protein to repress transcription.

MBC hydrolase gene (mheI)(BBa_K2244004) is derived from carbendazim-degrading strain Mycobacterium sp. SD-4 and is a genomic DNA gene sequence. mheI gene is also evolutionary conserved in many other bacteria strains, such as Nocardioides sp. SG-4G, and Rhodococcus erythropolis.

mCherry(BBa_K2244008) is a red fluorescent protein used as a reporter. It is based on a fluorescent protein that was originally isolated from Discosoma sp.mCherry sequence is codon optimized for E. coli expressionm and can be replaced by any gene sequence in light-regulated studies.

T1 terminator(BBa_B0010) is the most commonly used terminator.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1250
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 1250
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 754
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1250
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1250
  Illegal AgeI site found at 413
  
• 1000
  COMPATIBLE WITH RFC[1000]