Intermediate

Part:BBa_J31010:Design

Designed by: Karmella Haynes   Group: iGEM06_Davidson   (2006-10-28)
Revision as of 00:46, 29 October 2006 by Registry (Talk | contribs)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


KanB : RBSrev


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 383
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 664


Design Notes

Kan was PCR-amplified with primers designed to introduce a SpeI site to the left and an XbaI site to the right (reversing the normal order of these two BioBrick cut sites. The product was cut with SpeI and XbaI and ligated into S/X cut pSB1A2. The source for this vector was pSB1A2 carrying RFP. After cloning, we used a S/X digest to check for insertion in the desired oreitnation (reverse). Clones where Kan did not excise inserted in the wrong way (failure to cut is due to S/X mixed sites) and clones where Kan excised successfully inserted in the desired orientation (restoration of S and X sites).


Source

Parts BBa_J31002 and BBa_J44001

References