Plasmid_Backbone

Part:BBa_K260002:Design

Designed by: Kaj Bernhardt   Group: iGEM09_BIOTEC_Dresden   (2009-10-17)


pTetFlp. [anhydrotetracycline] -> FLP(F70L)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3222
    Illegal EcoRI site found at 5762
    Illegal XbaI site found at 67
    Illegal XbaI site found at 6532
    Illegal SpeI site found at 748
    Illegal SpeI site found at 5444
    Illegal PstI site found at 3132
    Illegal PstI site found at 4756
    Illegal PstI site found at 6283
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3222
    Illegal EcoRI site found at 5762
    Illegal EcoRI site found at 6680
    Illegal NheI site found at 1771
    Illegal SpeI site found at 2
    Illegal SpeI site found at 748
    Illegal SpeI site found at 5444
    Illegal PstI site found at 16
    Illegal PstI site found at 3132
    Illegal PstI site found at 4756
    Illegal PstI site found at 6283
    Illegal NotI site found at 9
    Illegal NotI site found at 6686
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3222
    Illegal EcoRI site found at 5762
    Illegal EcoRI site found at 6680
    Illegal BglII site found at 4969
    Illegal XhoI site found at 61
    Illegal XhoI site found at 6674
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 6680
    Illegal suffix found at 2
    Illegal EcoRI site found at 3222
    Illegal EcoRI site found at 5762
    Illegal XbaI site found at 67
    Illegal XbaI site found at 6532
    Illegal SpeI site found at 748
    Illegal SpeI site found at 5444
    Illegal PstI site found at 3132
    Illegal PstI site found at 4756
    Illegal PstI site found at 6283
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 6680
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3222
    Illegal EcoRI site found at 5762
    Illegal XbaI site found at 67
    Illegal XbaI site found at 6532
    Illegal XbaI site found at 6695
    Illegal SpeI site found at 2
    Illegal SpeI site found at 748
    Illegal SpeI site found at 5444
    Illegal PstI site found at 16
    Illegal PstI site found at 3132
    Illegal PstI site found at 4756
    Illegal PstI site found at 6283
    Illegal NgoMIV site found at 3927
    Illegal NgoMIV site found at 4305
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal SapI site found at 4245
    Illegal SapI site found at 4455


Design Notes

dhfr had to be deleted as the same gene for trimpethoprim resistance was going to be used elsewhere in the same cell.


Source

This plasmid comes from Sarov M, Schneider S, Pozniakovski A, Roguev A, Ernst S, Zhang Y, Hyman AA, Stewart AF. A recombineering pipeline for functional genomics applied to Caenorhabditis elegans. Nat Methods. 2006 Oct;3(10):839-44. However, it was modified by deletion of dhfr gene and RedABGRecA operon through insertion of a kanamycin resistance cassette.

References