Plasmid

Part:BBa_K4761428

Designed by: Pinhan Pan   Group: iGEM23_HiZJU-China   (2023-09-30)


pTarget-pykF4

SgRNA(pykF)4 expression plasmid for pykF knockout. Created based on sgRNA(pykF)4 (BBa_K4761028) and pTarget (BBa_K4761107) using overlapping PCR for guide RNA to function in CRISPR-Cas9 double plasmid system.

Fig.1 The sgRNA(pykF)4 expression box was successfully constructed. DNA sequencing results showed that sgRNA(pykF)4 was successfully inserted into the pTarget plasmid.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2065
    Illegal XbaI site found at 2071
    Illegal SpeI site found at 1955
    Illegal PstI site found at 1973
    Illegal PstI site found at 2083
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2065
    Illegal NheI site found at 1932
    Illegal SpeI site found at 1955
    Illegal PstI site found at 1973
    Illegal PstI site found at 2083
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2065
    Illegal BglII site found at 2095
    Illegal BamHI site found at 1920
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2065
    Illegal XbaI site found at 2071
    Illegal SpeI site found at 1955
    Illegal PstI site found at 1973
    Illegal PstI site found at 2083
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2065
    Illegal XbaI site found at 2071
    Illegal SpeI site found at 1955
    Illegal PstI site found at 1973
    Illegal PstI site found at 2083
    Illegal NgoMIV site found at 771
  • 1000
    COMPATIBLE WITH RFC[1000]


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