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 containing this part in a plasmid along with 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 resistance, growing on chloramphenicol + ampicillin. That B) and C) grow on these plates indicates that the cells contain both plasmids, and that the red colour has disappeared indicates that RFP is being actively repressed.