Device

Part:BBa_K3402056

Designed by: Yibo Shi   Group: iGEM20_Jiangnan_China   (2020-10-22)
Revision as of 04:40, 26 October 2020 by ZZ 0991 (Talk | contribs)


Single-gene editing cassette

This device is composed of 50bp-upPXA1(BBa_K3402037), hph(BBa_K3402012), 50bp-doPXA1(BBa_K3402038), Ptef1(BBa_K3402007), sgPXA1(BBa_K3402039), Tsyn7(BBa_K3402001).

Single-gene editing cassette sequence.png

Usage and Biology

We achieve the knockout of PXA1 gene and insert the hygromycin resistant gene as a selection marker. Two parts of PXA1 are homologous arms at the ends.
If modified strains can grow on the plate coated with hygromycin, it is successful to knock PXA1 with Cas9 system. By comparison, the editing efficiency of the CRISPR/Cas9 system can be clearly observed.
The single gene-editing efficiency was 100%.

The plates contained hygromycin


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 2171
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None