Difference between revisions of "Part:BBa K3924054"
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==Design and Construction== | ==Design and Construction== | ||
For this part, we can purchase directly. But to save time, we got it from Xing Lab, Tsinghua University. We use PCR to design this plasmid to contain our desired sgRNA plasmid | For this part, we can purchase directly. But to save time, we got it from Xing Lab, Tsinghua University. We use PCR to design this plasmid to contain our desired sgRNA plasmid | ||
| − | [[Image: | + | [[Image:T--Tsinghua--sgRNA.png|center|600px|thumb|'''Fig.1 Design for sgRNA part, the sgRNA is preceded by a constant promoter, J23119.''']] |
==Functional Verification== | ==Functional Verification== | ||
Latest revision as of 03:53, 22 October 2021
J23119-guideRNA-sgRNAscarffold terminator
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 30
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal SpeI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 30
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 30
- 1000COMPATIBLE WITH RFC[1000]
Profile
Name:J23119-guideRNA-sgRNAscarffold terminator
Base Pairs: 137bp
Origin: Escherichia coli
Properties: A constitutive promoter with a sgRNA DNA seuqence.
Usage and Biology
It constitutively encodes sgRNA to guide dCas9 into specific sites of a target gene
Design and Construction
For this part, we can purchase directly. But to save time, we got it from Xing Lab, Tsinghua University. We use PCR to design this plasmid to contain our desired sgRNA plasmid
Functional Verification
It has been reported functional ,but we failed to repeat it in our lab due to some reasons explained in Wiki.
Reference
[1]Banno S, Nishida K, Arazoe T, Mitsunobu H, Kondo A. Deaminase-mediated multiplex genome editing in Escherichia coli. Nat Microbiol. 2018;3(4):423-429. doi:10.1038/s41564-017-0102-6