Plasmid

Part:BBa_K4808009

Designed by: Zhao Guichun   Group: iGEM23_AIS-China   (2023-10-10)
Revision as of 14:29, 12 October 2023 by Gracezhao (Talk | contribs)

pTarget

pTarget plasmid that carrying specific gRNA sequence which can identity the target gene for the further gene knockout.

Characterization

pTarget plasmid play an important role in our CRISPR-CAS 9 knocking out experiment.

f49dac1bd8ad0ec8913a15418c6e2f4d.png

Step1: Transformation of the pEcCas plasmid(carry cas9 protein) into E. coli AIS-0 (CICC20905). Step2: construction of the pTarget plasmid and donor DNA. Step3: transformation of pTarget plasmid and donor DNA into the AIS-0 with pEsCas inserted. Step4: Incubation of the transformed strains.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 330
    Illegal XbaI site found at 336
    Illegal SpeI site found at 221
    Illegal PstI site found at 348
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 330
    Illegal NheI site found at 198
    Illegal SpeI site found at 221
    Illegal PstI site found at 348
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 330
    Illegal BglII site found at 360
    Illegal BamHI site found at 186
    Illegal XhoI site found at 384
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 330
    Illegal XbaI site found at 336
    Illegal SpeI site found at 221
    Illegal PstI site found at 348
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 330
    Illegal XbaI site found at 336
    Illegal SpeI site found at 221
    Illegal PstI site found at 348
    Illegal NgoMIV site found at 1155
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 122


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