Part:BBa_I739101

Double Promoter (constitutive / TetR, negative)

Part Structure

The Biobrick consists of the whole constitutive promoter BBa_J23100 linked to twice the first operator sequence of BBa_R0040 (TetR 1).

The -35 and -10 regions, which are responsible for the binding of RNA-polymerase to DNA via sigma factors, are included in the BBa_J23100 promoter region (first part of the double promoter). No further functional -10 and -35 regions were included in the second part of the double promoter. Instead, two operator sequences of the same type were included in the second part. The reason for this is that the probability of a binding event (and hence regulation strength) is increased if a multiple of identical operator sequences is included in the regulatory sequence.

Summarized:
There are three operator sequences: 1xBBa_J23100, 2xBBa_R0040 (TetR 1).

Mode of Action

The consitutive promoter J23100 is not regulated. However, when TetR binds to the TetR operator sequences, it represses transcription. If the inducer [http://openwetware.org/wiki/ATc anhydrotetracycline (ATc)] is added, TetR action is inhibited and the promoter gets derepressed.

Purpose

This Biobrick was designed for the [http://2007.igem.org/ETHZ ETHZ iGEM 2007 project] and belongs to a first generation of double promoters. This specific double promoter was originally designed as a proof of concept (PoC) promoter. The goal is to show the applicability of such kind of promoters. BBa_I739101 is part of biobrick BBa_I739014.

Testing

Checked for uniqueness of restriction enzyme cleavage sites:
Eco: ok
Xba: ok
Spe: ok
Pst: ok

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]