Difference between revisions of "Part:BBa K3081043"

Latest revision as of 19:20, 21 October 2019

CRISPR-based replication interference system for p15A plasmid copy number control, RNA I 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 template strand of replication inhibitor (RNA II) in p15A origin. Interestingly, it can upregulate the 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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE 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
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 1144
Illegal BamHI site found at 4600
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 979
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI site found at 961

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K3081043 short</partinfo>
-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 template strand of replication inhibitor (RNA II) in p15A origin. However, it shows no effect for some unknown reasons.
+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 template strand of replication inhibitor (RNA II) in p15A origin. Interestingly, it can upregulate the copy number..
 <center>
 https://2019.igem.org/wiki/images/c/ca/T--Peking--p15A_mechanism.png
@@ Line 15: / Line 15: @@
 <b>The effect of 4 target sites on p15A origin</b>
 </center>
+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.