Composite

Part:BBa_K1714003

Designed by: Masaya Mitsumoto   Group: iGEM15_HokkaidoU_Japan   (2015-09-13)
Revision as of 23:26, 18 September 2015 by Kakurehana (Talk | contribs)

pLac-RBS B0034-His10-TEV-thanatin-double terminator

This part is designed for thanatin expression system regulated by pLac and decreased the toxicity by adding His10. 

By adding IPTG, this part express His10 - TE - thanatin.
The His10 is also used for purification. After purification, they can be cleaved His10 and reactivated by TEV protease.

Even we subcultured many colonies from LB plate to liquid culture, the number of correctly subcultured products were few. We expected this result could be caused by thanatin's strong toxicity. Expect naturally modified cells, losing thanatin expression systems were not successed. It's very hard to survive with thanatin expression system even their toxicity was decreased by adding His10 and to get ideal sequences. We recommend you check whether this part's sequence is ideal sequence or not.

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 287
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None