Designed by: 2014 Ivy Tech South Bend IN team Group: iGEM14_IvyTech_SouthBendIN (2014-08-22)
T7 strong promoter + endolysin cassette
T7 strong promoter coupled with an endolysin generator designed to short circuit the replication cycle of T7 bacteriophage.
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 345 Illegal AgeI site found at 415
1000
COMPATIBLE WITH RFC[1000]
Characterization and Applications of BBa_K1477030
alt text
In correspondence with a 500 basepair DNA ladder, our part measures at ~2500 base pairs which corresponds with the sum of the BioBricks and the pSB1C3 backbone. Plasmid DNA isolated from two subclones
alt text
TEST FOR IMMEDIATE LYSIS OF TOP10 CELLS TRANSFORMED WITH BBa_K1477030.
This immediate-lysis device was designed to be an immediate lysis cassette for T7 infection of transformed Top10 cells. The expected result was that a culture of transformed cells would be resistant to infection by T7 phage but not by an unrelated phage. This experiment represents transformed cells infected at a ratio of 100 cells to 1 pfu T7 phage. If functioning, the immediate lysis cassette should have stopped the propagation of T7 infection. What may be lacking is the presence of holin that should increase the effectiveness of the endolysin.
