Part:BBa_K1444022

Repressible RFP

Constitutive high level expression of RFP, repressible by BBa_C0053.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1003
  Illegal AgeI site found at 1115
  
• 1000
  COMPATIBLE WITH RFC[1000]

This part is one of a series of designed repressors used by the 2014 Calgary iGEM team BBa_K1444006, cloned in front of RFP. It was designed to contain 3 copies of the repressible operator site BBa_R0053 in order to ensure tight and consistent repression.

Usage and Biology

Figure 1: Repression of RFP expression by C2-P22.

The series of plates shows that our repressor construct BBa_K1444023 is able to prevent expression of our reporter construct. All four plates are streaked with a strain of E. coli that contains a constitutive, but repressible, promoter in front of RFP, and resistance to chloramphenicol. A) shows that this strain grows on chloramphenicol and that it expresses RFP, and D) shows that this strain cannot grow on chloramphenicol + ampicillin. B) and C) are replicates that have been transformed with our repressor construct, which contains ampicillin. That B) and C) grow on chloramphenicol + ampicillin indicates that the cells contain both plasmids, and that the red colour has disappeared indicates that RFP is being actively repressed.