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Part:BBa_K1413041

Designed by: Sophia Belkhelfa   Group: iGEM14_Evry   (2014-10-07)

Mutation of OriVR6Kgamma origin of replication (ori)

This part is an origin of replication which needs pi protein to replicate. We used OriVR6Kgamma in BBa_K1413044 taken from pNK2.


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