Difference between revisions of "Part:BBa K2371012"
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− | In cell and molecular biology, the GFP gene is | + | 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 T7 report system in cell free system together with N-t7-dCas9(BBa_K2371000) and C-t7-dCas9(BBa_K2371001). | + | 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. | The user need to conduct a PCR to linearize the part before reacting with T7 report system. |
Revision as of 16:05, 31 October 2017
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.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 2.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:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 600
Illegal AgeI site found at 712 - 1000COMPATIBLE WITH RFC[1000]