Part:BBa_K2371012

pT7-amilGFP

In cell and molecular biology, the GFP gene is often used as reporter. We add a T7 promotor and a RBS in front of the GFP in order to improve it to be utilized in our paired dCas9 T7 report system in cell free system together with N-t7-dCas9(BBa_K2371000) and C-t7-dCas9(BBa_K2371001).

The user need to conduct a PCR to linearize the part before reacting with T7 report system.

We conducted both PCR check(VF and VR) and enzyme check(XbaI and SpeI)

Figure 1. The plasmid of pT7-amilGFP. pT7-amilGFP is composed of a T7 promotor, an RBS and the coding sequence of GFP. This plasmid is used to act as report gene in our report system.

Figure 2. PCR check of eforRed and amliGFP. The expected result of eforRed should be around 1000bp. The insufficient specificity of primers produce another band but the band around 1000bp can present the success of construction. The expected result of amliGFP should be around 1000bp. The control is J23119 biobrick.

Figure 3. RD check of eforRed and amliGFP. XbaI and SpeI were used. The expected length of both amliGFP and eforRed should be around 750bp.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 600
  Illegal AgeI site found at 712
  
• 1000
  COMPATIBLE WITH RFC[1000]