Composite

Part:BBa_K732002

Designed by: Joshua Fass   Group: iGEM12_Virginia   (2012-10-01)
Revision as of 04:29, 10 October 2012 by J.fass (Talk | contribs)

synCG-β + Promoter + RBS

This construct contains a strong constitutive promoter (J23119) as well as an RBS (B0034) upstream of synCG-β (K732001), and should strongly express a synthetic version of the beta subunit of human chorionic gonadotropin.

One application of this component is detection by pregnancy test.

Proof-of-principle

When heterogeneous mixture of E. coli expressing BBa_K732001 and BBa_K732002 was lysed by either sonication or T7 infection, re-folded using a literature-based protocol, and applied to NewChoice pregnancy tests (purchased from Dollar Tree), positive results were visible.

Note that (a) there was some coloration around the positive band, and (b) the pregnancy tests were left overnight for bands to become visible. We hypothesize that the delayed read-time is necessary because the folding solution interferes with band visibility.

Virginia 2012 Oct 2 Preg Test Results.jpg


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 224
    Illegal NgoMIV site found at 268
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//classic/generator/prc
//function/reporter
Parameters
None