Difference between revisions of "Part:BBa K4335021"

Revision as of 03:51, 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]

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.

Result

Verification of the effect of Golden Gate inserting specific sgRNA

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal PstI site found at 48
Illegal PstI site found at 121

12
INCOMPATIBLE WITH RFC[12]
Illegal PstI site found at 48
Illegal PstI site found at 121
Illegal NotI site found at 453

21
COMPATIBLE WITH RFC[21]

23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 48
Illegal PstI site found at 121

25
INCOMPATIBLE WITH RFC[25]
Illegal PstI site found at 48
Illegal PstI site found at 121

1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 551
Illegal BsaI.rc site found at 533

@@ Line 20: / Line 20: @@
 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>
 <br>
+<h2>Result</h2>
+<h3>Verification of the effect of Golden Gate inserting specific sgRNA<h3>
 </html>