Part:BBa_K1116002

TRE--LacI with T21 and miR-FF3 target

It was designed as a intermediate product to construct BBa_K1116003 in our project.

Characterization of BBa_K1116002

In our project, BBa_K1116002 serves as an auxiliary node. Our design as follows (Circuit A). Besides, BBa_K1116003 is used in our control design (Circuit B). The difference between BBa_K1116002 and BBa_K1116003 is the T21 targets.

By testing the mean value of EYFP (Enhance Yellow Fluorescent Protein) and the relationship between the EYFP and the DNA copy number, we can know whether the plasmids work or not. In our design, we use mkate (another fluorescent protein), which radiats red lights, as reference gene to reflect copy number. The results as follows. We can distinguish the relative valve of EYFP of Circuit A and Circuit B from the following figure.In the picture,1,2,3 are the results of Circuit A and 4,5,6 represent Circuit B.

We can know the miRNA-21 targets at T21,leading the decrease of gene lacI. So the expression of EYFP enhanced, thus the plasmid K1116002 works. Equally, we can prove the K1116003 is right. Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 381
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 381
  Illegal NotI site found at 1544
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 381
  Illegal BamHI site found at 343
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 381
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 381
  
• 1000
  COMPATIBLE WITH RFC[1000]