Device

Part:BBa_K3402060

Designed by: Yibo Shi   Group: iGEM20_Jiangnan_China   (2020-10-25)


Cas9 expression cassette without homologous arms

This device is composed of Ptef1 (BBa_K3402007), Cas9 (BBa_K3402023), NLS (BBa_K3402008) and Tsyn7 (BBa_K3402001).

Cas9 expression cassette without homologous arms.png

Usage and Biology

The use of the strongest promoter Ptef1 to express Cas9 protein and sgRNA could increase the gene-editing efficiency. As the Cas9 protein was too big to transport into the nucleus, a nuclear localization sequence (NLS) was needed to help the transportation of Cas9 protein.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 5675
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1730
    Illegal BglII site found at 5294
    Illegal XhoI site found at 1051
    Illegal XhoI site found at 1780
    Illegal XhoI site found at 2143
    Illegal XhoI site found at 5095
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 5675
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 5675
    Illegal NgoMIV site found at 2557
    Illegal NgoMIV site found at 3661
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1501
    Illegal BsaI site found at 2218


[edit]
Categories
Parameters
None