Part:BBa_I739021

Single regulated polycistronic P22 cII +LVA / EYFP +LVA expression cassette

Part Structure

The Biobrick encodes LVA-tagged P22 cII (BBa_C0053) and LVA-tagged EYFP (BBa_E0032) under control of the double promoter BBa_I739101. The ribosome binding sites BBa_B0034 are located upstream of each coding region. The transcription of P22 cII and EYFP is terminated by the double terminator BBa_B0015.

Mode of Action

P22 cII and EYFP production can be repressed by TetR. 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 is a composite biobrick including a first generation double promoter. The goal is to show the applicability of such kind of double promoters. In the project description, this part is a composite of the PoC intermediate and Part 5 and is cloned into plasmid pCK01BB1.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 275