Difference between revisions of "Part:BBa K1413041"

Latest revision as of 18:00, 2 November 2014

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

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1413041 short</partinfo>
-This part is an origin of replication which needs pi protein to replicate
+This part is an origin of replication which needs pi protein to replicate.
+We used OriVR6Kgamma in BBa_K1413044 taken from pNK2.
@@ Line 11: / Line 12: @@
 ===Sequence and Features===
 <partinfo>BBa_K1413041 SequenceAndFeatures</partinfo>
+This OriVR6Kgamma contained a mutation. The XbaI site present in the wild-type sequence was mutated to
+be RFC10 compatible. Using primers:
+<br/>Fw ORI Mut (transposas plasmid): GAGCCTTGAGACCTCTAGAGCAGACGTCATACTAGATAACTAGATC
+<br/>Rv ORI Mut: GACGCAGGAGACCACTAGTATAAGTGTTCCTGTGTCAC
 ===Functional Parameters===