Part:BBa_I721001

Lead Promoter

Lead Promoter (Part 4) messed up EcoRI site

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 SapI.rc site found at 80
  

Part BBa_1721001 is the Lead Promoter.

Description
This part is designed to enable Lead Detection in a cell. When Lead (Pb +2) enters the cell, it will couple with the Lead Binding protein (NOT IN THIS PART), forming a dimer. This dimer will then bind to the Lead Promoter, which is contained within this part, BBa_I721001. BBa_I721001 contains the Lead Promoter sequence, and and must be combined with a functional Lead Binding Protein part for it to work.

This sequence was obtained using PCR from the genome of Ralstonia metallidurans. Biobrick cut sites were added using specific primers during PCR.

This part is not known to function. "PbrR691/promoter combinations in presence of lead nitrate give no GFP production and no AHL production compared to control. "

The figure below illustrates the layout of this sequence.

Sequence Layout of I721003

This part contains one section: A sequence for a Lead Promoter.

15 planned Lead Promoter and Lead Binding Protein Biobricks

Because of the different coding regions contained in this sequence, the Brown iGEM Team chose 15 different regions of the Pbr691 and PbrR691 sequences to extract. We attempted to create Biobricks from each of these 15 different sequences. Some contained only the PbrR Lead Binding Protein, some contained only the Pbr Promoter, and some parts contained both. The following diagram illustrates these different parts.

BBa_I721001 contains the Lead Promoter region.

Functional Layout of I721003

BBa_I721001 was designed to be a part of the Brown University Cellular Lead detector device. The BBa_I721001 Lead Promoter is intended to be combined with a Lead Binding Protein Part. Together, this Lead Detection System was designed to be attached to the Amplification system by Imperial College, (BBa_J37015), with GFP as the output of the device. The schematic for this project is below:

Overall Brown University iGEM 07 Design