Device
"Lock"

Part:BBa_K318030

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

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

This composite part is a "lock" designed to prevent expression of a downstream gene unless a specific sequence of recombination events occurs to the part. Specifically, successive application of the Hin and Cre recombinases, respectively, will "unlock" the part leading to constitutive expression of the downstream gene(s). It was refered to as the "Lock Cassette" or "Lock Construct" by the [http://2010.igem.org/Team:Wisconsin-Madison Wisconsin-Madison 2010] iGEM team.

It can be used in conjunction with the Key Cassette after miscellaneous (junk) DNA is added to either the HindIII or the SapI (5' ATC 3' overhang) sites that are located within the part. According to the literature, it is necessary to have approximately 900 base pairs of DNA within the recombination sites for optimal recombination. As of October 2010, we were unable to characterize how the length of junk DNA affects the rates of recombination of the construct.

A zip file including information about this part is located here: File:BBa K318030 pSB1C3.zip

Sequence and Features


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


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Categories
Parameters
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