Difference between revisions of "Part:BBa K3365065"
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<partinfo>BBa_K3365065 short</partinfo> | <partinfo>BBa_K3365065 short</partinfo> | ||
| − | + | This part is a off-target detecting platform for dCas9-ω. We put this part in a plasmid and transfer it into △rpoZ-MG1655, a strain of E.coli that was koncked out rpoZ. If dCas9-ω is binding to this lure sequence, the submit Omega fused to dCas9 C-terminal can activate the expression of reporter by recruiting the RNAP holoenzyme and the cell would emit green fluorescence,which means dCas9-ωis off-target, else the weak promoter and the lack of rpoZ makes the reporter cannot express or just have a quite low expression level. | |
| + | |||
| + | According to the prediction model, the off-target rate of the lure sequence in this part is 0.37543%. | ||
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Revision as of 01:59, 25 October 2020
Fourth lure sequence upstream of eGFP
This part is a off-target detecting platform for dCas9-ω. We put this part in a plasmid and transfer it into △rpoZ-MG1655, a strain of E.coli that was koncked out rpoZ. If dCas9-ω is binding to this lure sequence, the submit Omega fused to dCas9 C-terminal can activate the expression of reporter by recruiting the RNAP holoenzyme and the cell would emit green fluorescence,which means dCas9-ωis off-target, else the weak promoter and the lack of rpoZ makes the reporter cannot express or just have a quite low expression level.
According to the prediction model, the off-target rate of the lure sequence in this part is 0.37543%.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 66
Illegal NheI site found at 89 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 834
Illegal XhoI site found at 843 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]