Composite

Part:BBa_K3156888

Designed by: Tingzhen Liu   Group: iGEM19_SHSBNU_China   (2019-10-16)
Revision as of 01:46, 22 October 2019 by Marshalliu (Talk | contribs) (Usage and Biology)


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).

Reference

[1]F. Farzadfard, T. K. Lu, Science 346,1256272 (2014). DOI: 10.1126/science.1256272

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 9
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1
    Illegal XhoI site found at 516
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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