Part:BBa_K3156888
pLac Promoter-ssDNA[sfGFP(ON)]-Ec86-Beta
Design
We combined msd-msr cassette, Reverse Transcriptase Ec-86 and Beta recombinase together to create BBa_K3156888. Msd-msr cassette can produce mRNA for reverse transcription to produce
Figure 1.Circuit design of BBa_K3156888(sfgfp forward).
We created a
Figure 2.Stimulated reparing progress of K3156888.
Usage and Biology
Culturing and plating
For each experiment, the samples were separately inoculated in LB medium plus appropriate antibiotics and grown overnight at 37°C, 200 revolutions per minute (RPM) to obtain seed cultures. Unless otherwise noted,inductions were performed by diluting the seed cultures (1:1000) in 2 ml of prewarmed LB plus appropriate antibiotics with or without inducers followed by 24 hours incubation 30°C, 700 RPM.
Figure 1.Repairing result of BBa_K3156888(sfgfp forward). 
Debug process
In the previous experiment, we found out that the 222# mutation of GFP on PSB4K5 plasmid could not be repaired successfully. The lumination of experimental group has no difference with the control group.Under this circumstance, we started our “debug” process.
Figure 1.Repairing result of BBa_K3156888(sfgfp reverse).
[1]F. Farzadfard, T. K. Lu, Science 346,1256272 (2014). DOI: 10.1126/science.1256272
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 9
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1
Illegal XhoI site found at 516 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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