Measurement

Part:BBa_K880001

Designed by: Josh Atkinson, Mike Ferguson, and Ben Parker   Group: iGEM12_Michigan   (2012-09-29)
Revision as of 00:19, 1 October 2012 by Jatk (Talk | contribs) (→‎A Utility for Assaying fim-Based Recombinases)

A Utility for Assaying fim-Based Recombinases

K880001 is a utility for characterizing the functionality of fim based recombinases such as FimB, FimE, and HbiF and consists of K137008 and K137010 flanking a “junk” DNA sequence K165006 containing an off-center AgeI restriction site proximal to the BioBrick prefix; inversion of the fragment will cause the site to move towards the suffix.

Asymmetrical_Digest_Reporter_Orientations.png

Digestion of the unflipped reporter with EcoRI and AgeI will yield a ~70bp fragment when in a FimE-flippable orientation and a ~220bp fragment when in an HbiF flippable orientation. Alternatively, digestion with AgeI and PstI will yield fragments of roughly opposite orientation. The reporter in the registry sequence is FimE flippable.

This is an artificial construct based off the assay performed in Teng et al. Infect. Immun. 2005, 73(5):2923.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 116
  • 1000
    COMPATIBLE WITH RFC[1000]
[edit]
Categories
Parameters
None