Part:BBa_K1021020

PT7+GFP

The gene for green fluorescent protein sits downstream of the T7 constitutive promoter. This gene can be naturally expressed in Escherichia coli BL21-A1 in conjunction with T7 polymerase and can be expressed in Escherichia coli DH5α after co-transformation.

Usage and Biology

• The part was constructed from BBa_I712074 and BBa_E0040.
• This construct was modified to contain a bacterial RBS so that we could characterize its transcriptional functionality. A fluorescence assay was run using a 96 well plate as described in the protocol section. T7+RBS+GFP and T7+GFP (BBa_K1021020) constructs in E. coli BL21-A1 were examined for their relative fluorescence (fluorescence/OD600). A T7 construct in E. coli BL21-A1 without downstream elements was utilized as a control along with the use of the aforementioned constructs in E. coli DH5α. As can be seen from the graph there were evident differences in peak fluorescence between the T7+RBS+GFP construct and the T7+GFP construct and controls. The T7+GFP+RBS construct appears to have a 20 fold higher relative fluorescence than the T7+GFP construct or the controls.

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
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 696