Difference between revisions of "Part:BBa K3081057"
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<b>The effect of 4 target sites on p15A origin</b> | <b>The effect of 4 target sites on p15A origin</b> | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
Latest revision as of 19:18, 21 October 2019
CRISPR-based replication interference system for p15A plasmid copy number control, ini(+) site
This composite part uses a CRISPR-based DNA replication interference system for the copy number control of the p15A plasmid. The dCas9 is expressed in an inducible manner with a ssrA degradation tag fused to its C-terminal to minimize the basal expression. The sgRNA has a 20-bp complementary sequence with the bottom strand (the strand that replication activator (RNA II) binds to) of initiation site (+1) on p15A origin. It can efficiently downregulate the plasmid copy number.
Design of the p15A plasmid copy number control system
The effect of 4 target sites on p15A origin
Reference：
[1]Wolański M, Donczew R, Zawilakpawlik A, et al. oriC-encoded instructions for the initiation of bacterial chromosome replication.[J]. Frontiers in Microbiology, 2015, 5(735):735.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1205
Illegal NheI site found at 2321
Illegal NheI site found at 5495
Illegal NheI site found at 5518 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
Illegal BamHI site found at 4600 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 979
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961