Plasmid_Backbone

Part:BBa_K260003:Design

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


pRhaFlp. [L-rhamnose] -> FLP(F70L)


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 3471
    Illegal EcoRI site found at 4272
    Illegal SpeI site found at 1125
    Illegal SpeI site found at 3153
    Illegal PstI site found at 2466
    Illegal PstI site found at 3992
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3471
    Illegal EcoRI site found at 4272
    Illegal EcoRI site found at 6324
    Illegal NheI site found at 102
    Illegal NheI site found at 5981
    Illegal SpeI site found at 2
    Illegal SpeI site found at 1125
    Illegal SpeI site found at 3153
    Illegal PstI site found at 16
    Illegal PstI site found at 2466
    Illegal PstI site found at 3992
    Illegal NotI site found at 9
    Illegal NotI site found at 6330
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3471
    Illegal EcoRI site found at 4272
    Illegal EcoRI site found at 6324
    Illegal BglII site found at 2679
    Illegal BglII site found at 4952
    Illegal XhoI site found at 61
    Illegal XhoI site found at 6318
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 6324
    Illegal suffix found at 2
    Illegal EcoRI site found at 3471
    Illegal EcoRI site found at 4272
    Illegal SpeI site found at 1125
    Illegal SpeI site found at 3153
    Illegal PstI site found at 2466
    Illegal PstI site found at 3992
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 6324
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3471
    Illegal EcoRI site found at 4272
    Illegal XbaI site found at 6339
    Illegal SpeI site found at 2
    Illegal SpeI site found at 1125
    Illegal SpeI site found at 3153
    Illegal PstI site found at 16
    Illegal PstI site found at 2466
    Illegal PstI site found at 3992
    Illegal NgoMIV site found at 2015
    Illegal AgeI site found at 4684
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal SapI site found at 1955
    Illegal SapI site found at 2165


Design Notes

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.


Source

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

References