Plasmid

Part:BBa_K3385001:Design

Designed by: Daniel Bavnh�j   Group: iGEM20_DTU-Denmark   (2020-10-12)


pFC902


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1720
    Illegal EcoRI site found at 3997
    Illegal EcoRI site found at 6210
    Illegal EcoRI site found at 6361
    Illegal EcoRI site found at 8634
    Illegal EcoRI site found at 8897
    Illegal EcoRI site found at 10542
    Illegal XbaI site found at 665
    Illegal XbaI site found at 1147
    Illegal XbaI site found at 1915
    Illegal XbaI site found at 3802
    Illegal XbaI site found at 4570
    Illegal XbaI site found at 5052
    Illegal XbaI site found at 12197
    Illegal SpeI site found at 82
    Illegal SpeI site found at 93
    Illegal SpeI site found at 457
    Illegal SpeI site found at 589
    Illegal SpeI site found at 713
    Illegal SpeI site found at 820
    Illegal SpeI site found at 831
    Illegal PstI site found at 2864
    Illegal PstI site found at 13001
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1720
    Illegal EcoRI site found at 3997
    Illegal EcoRI site found at 6210
    Illegal EcoRI site found at 6361
    Illegal EcoRI site found at 8634
    Illegal EcoRI site found at 8897
    Illegal EcoRI site found at 10542
    Illegal NheI site found at 6054
    Illegal NheI site found at 13657
    Illegal SpeI site found at 82
    Illegal SpeI site found at 93
    Illegal SpeI site found at 457
    Illegal SpeI site found at 589
    Illegal SpeI site found at 713
    Illegal SpeI site found at 820
    Illegal SpeI site found at 831
    Illegal PstI site found at 2864
    Illegal PstI site found at 13001
    Illegal NotI site found at 13648
    Illegal NotI site found at 13726
    Illegal NotI site found at 16552
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1720
    Illegal EcoRI site found at 3997
    Illegal EcoRI site found at 6210
    Illegal EcoRI site found at 6361
    Illegal EcoRI site found at 8634
    Illegal EcoRI site found at 8897
    Illegal EcoRI site found at 10542
    Illegal BglII site found at 11792
    Illegal BamHI site found at 9708
    Illegal BamHI site found at 12052
    Illegal BamHI site found at 12203
    Illegal BamHI site found at 12739
    Illegal XhoI site found at 2261
    Illegal XhoI site found at 2978
    Illegal XhoI site found at 3456
    Illegal XhoI site found at 11067
    Illegal XhoI site found at 12599
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1720
    Illegal EcoRI site found at 3997
    Illegal EcoRI site found at 6210
    Illegal EcoRI site found at 6361
    Illegal EcoRI site found at 8634
    Illegal EcoRI site found at 8897
    Illegal EcoRI site found at 10542
    Illegal XbaI site found at 665
    Illegal XbaI site found at 1147
    Illegal XbaI site found at 1915
    Illegal XbaI site found at 3802
    Illegal XbaI site found at 4570
    Illegal XbaI site found at 5052
    Illegal XbaI site found at 12197
    Illegal SpeI site found at 82
    Illegal SpeI site found at 93
    Illegal SpeI site found at 457
    Illegal SpeI site found at 589
    Illegal SpeI site found at 713
    Illegal SpeI site found at 820
    Illegal SpeI site found at 831
    Illegal PstI site found at 2864
    Illegal PstI site found at 13001
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1720
    Illegal EcoRI site found at 3997
    Illegal EcoRI site found at 6210
    Illegal EcoRI site found at 6361
    Illegal EcoRI site found at 8634
    Illegal EcoRI site found at 8897
    Illegal EcoRI site found at 10542
    Illegal XbaI site found at 665
    Illegal XbaI site found at 1147
    Illegal XbaI site found at 1915
    Illegal XbaI site found at 3802
    Illegal XbaI site found at 4570
    Illegal XbaI site found at 5052
    Illegal XbaI site found at 12197
    Illegal SpeI site found at 82
    Illegal SpeI site found at 93
    Illegal SpeI site found at 457
    Illegal SpeI site found at 589
    Illegal SpeI site found at 713
    Illegal SpeI site found at 820
    Illegal SpeI site found at 831
    Illegal PstI site found at 2864
    Illegal PstI site found at 13001
    Illegal NgoMIV site found at 2998
    Illegal NgoMIV site found at 7729
    Illegal AgeI site found at 5925
    Illegal AgeI site found at 10015
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 5591
    Illegal BsaI site found at 12792
    Illegal BsaI.rc site found at 126
    Illegal BsaI.rc site found at 5883
    Illegal BsaI.rc site found at 7692
    Illegal BsaI.rc site found at 10713
    Illegal SapI site found at 7676
    Illegal SapI site found at 8072


Design Notes

The amount of gRNA's has to be considered, as it is possible to use 1 or more gRNAs, as long as they have overhangs to fit each other and fit into the PacI/Nt.BbvCI cassette for integration into the pFC330 plasmid.

Source

This part is assembled from different parts obtained from other organisms, such as the Streptococcus pyogenes viral defense protein Cas9 and pyrG plus AMA1 from other Aspergillus species, optimized for A. niger[1].

pFC902 was a gift from Uffe Mortensen (Addgene plasmid # 106904; http://n2t.net/addgene:106904; RRID:Addgene_106904)

References

[1] Efficient Oligo nucleotide mediated CRISPR-Cas9 Gene Editing in Aspergilli. Nodvig CS, Hoof JB, Kogle ME, Jarczynska ZD, Lehmbeck J, Klitgaard DK, Mortensen UH. Fungal Genet Biol. 2018 Jan 8. pii: S1087-1845(18)30004-5. doi: 10.1016/j.fgb.2018.01.004. 10.1016/j.fgb.2018.01.004 PubMed 29325827