Difference between revisions of "Part:BBa K4335021"
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<h2>Usage</h2> | <h2>Usage</h2> | ||
pCrU6.3+insert site+SpScaffold+polyT Term is one of the core functional components of plasmid pTX2038 and pTX2040 Its main function is to express specific sgRNA, and combine with | pCrU6.3+insert site+SpScaffold+polyT Term is one of the core functional components of plasmid pTX2038 and pTX2040 Its main function is to express specific sgRNA, and combine with | ||
| − | HSP70A Promoter+3×Flag+NLS+SpCas9+NLS+Rbcs2 Term <a href="https://parts.igem.org/Part:BBa_K4335020">[BBa_K4335020]</a>works together to cut specific genes in the nuclear genome of Chlamydomonas reinhardtii, so as to knock out negative lipid related regulatory genes. | + | HSP70A Promoter+3×Flag+NLS+SpCas9+NLS+Rbcs2 Term <a href="https://parts.igem.org/Part:BBa_K4335020">[BBa_K4335020]</a>works together to cut specific genes in the nuclear genome of Chlamydomonas reinhardtii, so as to knock out negative lipid related regulatory genes.<br> |
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Revision as of 03:49, 11 October 2022
pCrU6.3+insert site+SpScaffold+polyT Term
BBa_ K4335021 consists of a terminator, a cloning site, SpScaffold and a terminator
PCrU6.3 is a previously characterized RNA from chromosome 8 of Chlamydomonas reinhardtii [1] sgRNA
Template of sgRNA. Use Golden Gate (BsaⅠ) to replace the template sequence with a ~20bp guide sequence. Add 'ACTT' on the 5'-terminal of your guide sequence and 'AAAC' on the reverse 5'-terminal.
Usage
pCrU6.3+insert site+SpScaffold+polyT Term is one of the core functional components of plasmid pTX2038 and pTX2040 Its main function is to express specific sgRNA, and combine with HSP70A Promoter+3×Flag+NLS+SpCas9+NLS+Rbcs2 Term [BBa_K4335020]works together to cut specific genes in the nuclear genome of Chlamydomonas reinhardtii, so as to knock out negative lipid related regulatory genes.Sequence and Features