Difference between revisions of "Part:BBa K3365001"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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It is the target binding site for dCas9 fused with subnit Omega in transcription acvating system. | It is the target binding site for dCas9 fused with subnit Omega in transcription acvating system. | ||
Revision as of 12:31, 27 October 2020
Target binding site for dCas9 and subnit Omega
This part is used to detect whether dCas9 protein is on-target. The sequence is made up of PDCD1, which is the binding site of dCas9, and J23117 BBa_J23117 (visit:[1]) weak promoter spaced by 35 base pairs. The dCas9 in this part could bind to the PDCD1, under guidance of specific designed sgRNA, while subunit Omega could bind to J23117 and activating the expression of downstream fluorescent protein. According to reference, we knew that 35 base pairs is the best length of spacer for activating, so we randomly choose 35bp sequence from the plasmid pUC57 as spacer.
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 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]