Part:BBa_K1639009

LacI regulated DsRed with miR 26a and miR 375 binding sites

Usage and Biology

In our project Cancer module consist of three parts. First one produces tTA to activate expression of mLacI protein form pTRE. "m" in mLacI stands for mammalian, it's optimized version of bacterial LacI for expression in eucaryots.Produced mLacI would suppress CMV IE promoter through lac operators.(Figure 1)

File:Cacner.png

Figure 1:A detailed diagram of miRNA recognition system

This part composed of two lac operators which followed up with DsRed protein (red flourescent protein derived from Discosoma spp) and miRNA binding sites at 3'end. This miRNA binding sites are complementary to low-miRNA in gastric cancer cells. In normal cells the level of these miRNas are higher than in gastric cancerous cells they would suppress expression of DsRed protein.

Cloning and Expression

Sequence and Features

Assembly Compatibility:

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

  Illegal NotI site found at 835
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 593
  Illegal BamHI site found at 876
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 535
  
• 1000
  COMPATIBLE WITH RFC[1000]