Regulatory
Part:BBa_K4335012
Designed by: Hang Zhang Group: iGEM22_UESTC-BioTech (2022-10-10)
pCrU6.3
pCrU6.3 is a previously characterized RNA polymerase III promoter from the chromosome 8 of Chlamydomonas reinhardtii
The first prerequisite for CRISPR/Cas9 experiments is sgRNA transcription under the control of the RNA polymerase III promoter (RNAPII) . In general, the RNAPII transcription factor binding site is intrinsic to the transcriptional sequence and therefore not amenable to sgRNA transcription. U6snRNA is one of the few examples of sgRNAs with proximal RNAPII promoter elements. Therefore, we used previously characterized U6snRNA sequences from chromosome 8.
Usage
pCrU6.3 was constructed into plasmid pTX2038 and pTX2040 as the promoter of gRNA, and was successfully introduced into Chlamydomonas reinhardtii to obtain the positive transformant.Discription
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 48
Illegal PstI site found at 121 - 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 48
Illegal PstI site found at 121
Illegal NotI site found at 453 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 48
Illegal PstI site found at 121 - 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 48
Illegal PstI site found at 121 - 1000COMPATIBLE WITH RFC[1000]
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Categories
Parameters
None |