Part:BBa_I739002

Constitutive expression cassette for LacI +LVA (J23100.B0034.C0012.B0015)

Part Structure

The Biobrick encodes LVA-tagged LacI (BBa_C0012) under control of the constitutive promoter BBa_J23100 followed by the ribosome binding site BBa_B0034. The transcription of lacI is terminated by the double terminator BBa_B0015.

Mode of Action

LacI binds to the pLac regulator (BBa_R0010) (or to the PLlac01 hybrid regulator BBa_R0011 respectively) and 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://parts.mit.edu/igem07/index.php/ETHZ ETHZ iGEM 2007 project] and belongs to the constitutive part of the system. In the project description, this part is also termed Part 2. The constitutively synthesized LacI interacts with the double promoters BBa_I739103 and BBa_I739107 which are parts of composites BBa_I739016 and BBa_I739011 respectively. A similar (but not used) construct in this context is BBa_Q04121.

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
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1208
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]