Composite

Part:BBa_K1081000

Designed by: Hangxing Jia   Group: iGEM13_WHU-China   (2013-09-10)

J13002-dcas9


Cas9 is an RNA-guided double-stranded DNA endonuclease from Streptococcus pyogenes. Here we use dCas9(“dead” Cas9) ,in which two mutations of D10A and H840A were introduced to abolish the endonuclease function.

We combine dcas9 with different guide-RNAs to target different regions of double-promoter which we created. In this way, we can accomplish different expression level of the target genes.

In BBa_K1081000, we add J13002, which is composed of p(tetR) R0040 and RBS BOO34, before dcas9 to make dcas9 protein expression controllable.


Sequence and Features

Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1175
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 3454
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//function/crispr
//function/crispr/cas9
Parameters
None