Composite

Part:BBa_K3044023

Designed by: Catharina Bang Jensen   Group: iGEM19_SDU-Denmark   (2019-10-07)
Revision as of 23:57, 7 October 2019 by Catharina (Talk | contribs)


Conjugative sgRNA-dCas9 system

This dCas9 is a codon optimized version of BBa_K1150000 for expression in E. coli. Together with a single guide RNA (sgRNA), the protein can be used for silencing of specific target genes. dCas9 is the inactive form of the endonuclease Cas9. In contrast to Cas9, dCas9 does not cleave the DNA. Instead, it sterically blocks the elongation of transcription as a result of the sgRNA-DNA interaction. This will only repress the expression of the target gene thus avoiding the destruction of the gene. [1]

The dCas9 is asembled with the constitutive active promoter BBa_J23100, an RBS BBa_B0034 and a terminator BBa_B0010.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 478
    Illegal BglII site found at 1552
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 4547
    Illegal NgoMIV site found at 4557
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 4449


[edit]
Categories
Parameters
None