Part:BBa_K3995004

Pprovoin5_amilGFP

Pprovoin5_amilGFP

Usage and Biology

Ptat_AtzR is a coding sequence for expressing atzR. Cyanuric acid combination with AtzR can activate amilGFP expression to show the signs of fluorescence. The content of cyanuric acid is detected by detecting the fluorescence concentration. So Ptat_AtzR should be used with Pprovoin5_amilGFP.

Figure1.Genetic design of biosensor.

Construct Design

amilGFP is key functional factors that show the signs of fluorescence which is controlled by a Pprovoin5 promoter (Figure 2). The Pprovoin5_amilGFP is inserted in the pUC57 mini vector to get plasmid A (Figure 3).

Figure 2. Pprovoin5_amilGFP box..

Figure 3. Schematic maps of Pprovoin5_amilGFP (reporter plasmid)..

BBa_K3995002

Usage and Biology

This operator region has a leftward-facing repressible promoter and a rightward-facing activated promoter. AtzR remains bound to the DNA. In the natural system, the left-facing promoter is upstream of the atzR CDS, and atzR is autoregulatory; in the presence of cyanuric acid, the binding of AtzR changes to cause the right-facing promoter to become activated, allowing for the expression of the downstream atzDEF CDS. The presence of cyanuric acid "frees" AtzR from being bound to the atzDEF promoter so polymerase can now bind and transcribe following gene. AtzR remains bound to the atzR promoter region even in the presence of cyanuric acid. Note that high glnK concentrations are also needed, as glnK is a co-activator of downstream operator.

Lane pUC57-Pprovoin5: Plasmid pUC57-kana-mini-Pprovoin5 (2097bp) digested by Bsa1.

Sequence and Features

Assembly Compatibility:

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

  Illegal XhoI site found at 92
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]