Difference between revisions of "Part:BBa K2145102"
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This plasmid contains a GFP and an RFP reporter separated by a 282-bp spacer containing three gRNA binding sites for dCas. The reporter is designed to demonstrate the effect, if any, of binding a dCas "clamp" between two genes (the idea being that such a clamp can limit propagation of supercoiling between the genes, functionally isolating them). | This plasmid contains a GFP and an RFP reporter separated by a 282-bp spacer containing three gRNA binding sites for dCas. The reporter is designed to demonstrate the effect, if any, of binding a dCas "clamp" between two genes (the idea being that such a clamp can limit propagation of supercoiling between the genes, functionally isolating them). | ||
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| + | This part is one of a 4-plasmid set (K2145100, K2145101, K2145102, and K2145103). Other plasmids in this set are similar, but have GFP and RFP expression cassettes pointing in different directions. Different arrangement and orientations are expected to generate different patterns of supercoiling, which translates to different patterns of expression in RFP and GFP if supercoiling creates interference between the two. Addition of dCas and gRNAs matching the clamp sites are expected to reduce supercoiling, leading to more similar expression between the plasmids in this set. | ||
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Revision as of 23:39, 30 October 2016
GFP/RFP reporter (Forward/Reverse) separated by three dCas clamp sites
This plasmid contains a GFP and an RFP reporter separated by a 282-bp spacer containing three gRNA binding sites for dCas. The reporter is designed to demonstrate the effect, if any, of binding a dCas "clamp" between two genes (the idea being that such a clamp can limit propagation of supercoiling between the genes, functionally isolating them).
This part is one of a 4-plasmid set (K2145100, K2145101, K2145102, and K2145103). Other plasmids in this set are similar, but have GFP and RFP expression cassettes pointing in different directions. Different arrangement and orientations are expected to generate different patterns of supercoiling, which translates to different patterns of expression in RFP and GFP if supercoiling creates interference between the two. Addition of dCas and gRNAs matching the clamp sites are expected to reduce supercoiling, leading to more similar expression between the plasmids in this set.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1411
Illegal AgeI site found at 1523 - 1000COMPATIBLE WITH RFC[1000]