Part:BBa_K2016000

Strong constitutive E. coli promoter with included RBS - ready for cloning

BBa_K2016000 is a derivative of Bba_J23100, a promoter from a family of constitutive E. coli promoters isolated from a small combinatorial library. This family was submitted to the registry by Berkeley 2006. Sheffield 2016 has improved Bba_J23100 by adding an RBS downstream of the promoter to make it suitable for direct cloning. BBa_K2016000 is 91bp long (Fig. 1), first 35 nucleotides make up a functional strong constitutive promoter for expression in E. coli, followed by a 50 nucleotide spacer (5' UTR) and ending with a 6 nucleotide ribosomal binding site (AGGAGG).

Usage and Biology

Sheffield 2016 has used this promoter in order to design a biological device that responds to intracellular iron levels in bacteria. Bba_J23106 was also used in order to observe a difference between having a system under the control of a medium and a strong promoter. In the figure below we can see differences in the GFP fluorescence when expressed using a medium promoter (J23106 derivative) or strong promoter (J23100 derivative) in two different strains: W3110 (WT) and JC28 (∆entC)

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 38