Part:BBa_I739107

Double Promoter (cI, negative / LacI, negative)

Part Structure

The Biobrick consists of a large part of the whole non-constitutive promoter BBa_R051 (Basepairs 1-43) linked to the second operator sequence of BBa_R0010 (LacI binding site).

The -35 and -10 regions, which are responsible for the binding of RNA-polymerase to DNA via sigma factors, are included in the BBa_R0051 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.

Summarized:
There are three operator sequences: 1xBBa_R0051 (OR 2), 1xBBa_R0051 (OR 1), 1xBBa_R0010 (LacI binding site).

Mode of Action

Regulation of the first part: When lambda cI binds to the cI operator sequences, it represses transcription. There is no inducer which derepresses the promoter upon addition.

Regulation of the second part: When LacI binds to the LacI operator sequence, it represses transcription. If the inducer [http://openwetware.org/wiki/IPTG Isopropyl-beta-D-thiogalactopyranoside (IPTG)] is added, LacI 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. BBa_I739107 is part of biobrick BBa_I739011.

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
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]