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Part:BBa_K784043

Designed by: Inbal Vaknin   Group: iGEM12_Technion   (2012-09-25)
Revision as of 13:01, 25 September 2012 by Sl.ilya (Talk | contribs)

T7*(K1F) RNA polymerase

This part includes an engineered T7 RNAP with a RBS, the RNAP recognizes specifically a suitable promoter (pT7). The mutant T7 RNAP was donated us by Chris Voight. The T7 RNAP part contains a weak ribosome binding site and 'GTG' start codon . The native T7 RNAP differs from the mutant one by its toxicity for the host cell- the mutant have a low toxicity than the native one (according to Voight's article), this treat for low toxicity was accrued by a Spontaneous mutation, within the active site, during the cloning and doesn’t reduce the RNAP activity. The part was made by cloning the T7 RNAP gene to the BioBrick BBa_B0015, which includes double terminator (B0010-B0012). This part is present in the plasmid pSB1AK3. The purpose of this part is that you can clone to it any gene you want to be combined with it: promoter, reporter gene etc..
Note: the biobrick prefix has been altered to deletion of one bp downstream to the XbaI site. This was due to wrong primers.

Usage and Biology

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
Parameters
None