Device
"Lock"

Part:BBa_K318030:Design

Designed by: Justin Vrana, Nate Pantalone   Group: iGEM10_Wisconsin-Madison   (2010-07-13)

lox66 + rpCons + hixC + lox71 + SapI + T + T + Hin enhancer + HindIII + hixC


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 43
    Illegal NheI site found at 66
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 162


Design Notes

Note:

  • The constitutive promoter is oriented to express whatever gene is downstream of the final hixC site after recombination.
  • The SapI site located within the part yields a cut site as shown 5'(A\ATCT)3' on top and 3'(TTAG\A)5' on bottom.
  • The HindIII site located within the part actually allows for "lock breaking" because inserting a promoter into the part through standard cloning protocols bypasses the "lock" effect. The site was added to more easily recognize different recombinants through restriction digests.

Source

From synthesis but the same construct can be made with biobricks.

References