Composite

Part:BBa_K4687044

Designed by: Yiming Jiang   Group: iGEM23_HBUT-China   (2023-10-07)


MAD7+recE/T+pJYS1+PlacM:MADE/TJlacM-g5d3

The exonuclease–recombinase pair recE/T can improve dsDNA recombination efficiency in C. glutamicum .In this study, we combined the recE/T and CRISPR-MAD7 systems.Using this system, we optimized the CRISPR-MAD7 promoter sequence and screened for more efficient vector backbone and PAM recognition sites, and designed donor DNA to achieve targeted gene knockout in Corynebacterium glutamicum.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 9481
    Illegal EcoRI site found at 9724
    Illegal EcoRI site found at 10600
    Illegal EcoRI site found at 15996
    Illegal XbaI site found at 8698
    Illegal SpeI site found at 6464
    Illegal SpeI site found at 14418
    Illegal SpeI site found at 14797
    Illegal PstI site found at 7654
    Illegal PstI site found at 8944
    Illegal PstI site found at 9211
    Illegal PstI site found at 10405
    Illegal PstI site found at 12574
    Illegal PstI site found at 15500
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 9481
    Illegal EcoRI site found at 9724
    Illegal EcoRI site found at 10600
    Illegal EcoRI site found at 15996
    Illegal NheI site found at 15113
    Illegal SpeI site found at 6464
    Illegal SpeI site found at 14418
    Illegal SpeI site found at 14797
    Illegal PstI site found at 7654
    Illegal PstI site found at 8944
    Illegal PstI site found at 9211
    Illegal PstI site found at 10405
    Illegal PstI site found at 12574
    Illegal PstI site found at 15500
    Illegal NotI site found at 9811
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 9481
    Illegal EcoRI site found at 9724
    Illegal EcoRI site found at 10600
    Illegal EcoRI site found at 15996
    Illegal BglII site found at 714
    Illegal BglII site found at 762
    Illegal BglII site found at 1083
    Illegal BglII site found at 2298
    Illegal BglII site found at 2950
    Illegal BglII site found at 3804
    Illegal BglII site found at 8507
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 9481
    Illegal EcoRI site found at 9724
    Illegal EcoRI site found at 10600
    Illegal EcoRI site found at 15996
    Illegal XbaI site found at 8698
    Illegal SpeI site found at 6464
    Illegal SpeI site found at 14418
    Illegal SpeI site found at 14797
    Illegal PstI site found at 7654
    Illegal PstI site found at 8944
    Illegal PstI site found at 9211
    Illegal PstI site found at 10405
    Illegal PstI site found at 12574
    Illegal PstI site found at 15500
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 9481
    Illegal EcoRI site found at 9724
    Illegal EcoRI site found at 10600
    Illegal EcoRI site found at 15996
    Illegal XbaI site found at 8698
    Illegal SpeI site found at 6464
    Illegal SpeI site found at 14418
    Illegal SpeI site found at 14797
    Illegal PstI site found at 7654
    Illegal PstI site found at 8944
    Illegal PstI site found at 9211
    Illegal PstI site found at 10405
    Illegal PstI site found at 12574
    Illegal PstI site found at 15500
    Illegal NgoMIV site found at 5134
    Illegal NgoMIV site found at 13112
    Illegal NgoMIV site found at 16411
    Illegal AgeI site found at 1865
  • 1000
    COMPATIBLE WITH RFC[1000]


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