Difference between revisions of "Part:BBa K4335021"
Line 21: Line 21:
 
<br>
 
<br>
  
−
<html>
+
</html>
  
  
Line 31: Line 31:
 
<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 +
 
<partinfo>BBa_K4335021 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4335021 SequenceAndFeatures</partinfo>
  

Revision as of 03:50, 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.

sgRNA

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


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