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Part:BBa_K1413041

Designed by: Sophia Belkhelfa   Group: iGEM14_Evry   (2014-10-07)
Revision as of 21:02, 1 November 2014 by SophiaB (Talk | contribs) (→‎Sequence and Features)

Mutation of OriVR6Kgamma origin of replication (ori)

This part is an origin of replication which needs pi protein to replicate


Usage and Biology

It can be used in combinaison with transposon system which does not required a replicative system like in the part BBa_K1413044 .

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]

This OriVR6Kgamma contained a mutation. The XbaI site present in the wild-type sequence was mutated to be RFC10 compatible. Using primers:
Fw ORI Mut (transposas plasmid): GAGCCTTGAGACCTCTAGAGCAGACGTCATACTAGATAACTAGATC
Rv ORI Mut: GACGCAGGAGACCACTAGTATAAGTGTTCCTGTGTCAC

Functional Parameters

This part requires production of pi protein from pir gene.

References

Rakowski SA, Filutowicz;Plasmid R6K Replication Control; M. Plasmid. 2013 May; 69(3)231-242

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Categories
Parameters
None