Part:BBa_K2027015

Internal Ribosome Entry Site Transcriptional Regulation Mechanism-Theophylline Aptamer

Internal Ribosome Entry Site Transcriptional Regulation Mechanism-Theophylline Aptamer

Usage and Biology

IRES stands for Internal Ribosome Entry Site, and it does exactly what the name implies: allow protein transcription and thus transition to begin significantly downstream of the initial promoter. IRES sites are somewhat sensitive to 3D confirmations, and will only work when folded correctly. In this brick, we have attempted to assemble a small-molecule sensor system based on that property.

This brick is directly modeled after the system described in the following reference paper in 2011:

http://rnajournal.cshlp.org/content/17/3/478.long

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 BsaI.rc site found at 38
  Illegal BsaI.rc site found at 99
  Illegal BsaI.rc site found at 954
  

The part consists, in order, of: the iGEM prefix, an iGEM constitutive promoter sequence (BBa_BBa_J23119), a 5' stabilizing loop, an optimized modulator region, the anti-anti-IRES sequence (aaIRES), the aptamer that binds the molecule of interest, anti-IRES (aIRES), the IRES sequence, the translation start site, and then GFP (BBa_E0040). The aptamer domain is placed such that only the bound confirmation obscures the IRES-blocking site, thus allowing GFP translation to occur.