Part:BBa_K1116000

LacI with miR-21 and miR-FF3 target

This part is a LacI coding sequence with 4 miR-21 and 4 miR-FF3 target sites so it can be regulated with micro-RNA 21 and FF3. To test this part, we construct K1116002 and here is the data by which we characterized this part. You can also see it on the K1116002's page [[1]].

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

  Illegal NotI site found at 1120
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]