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: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==
 
It has been reported functional ,but we failed to repeat it in our lab due to some reasons explained in Wiki.
 
It has been reported functional ,but we failed to repeat it in our lab due to some reasons explained in Wiki.

Latest revision as of 03:53, 22 October 2021


J23119-guideRNA-sgRNAscarffold terminator


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal SpeI site found at 30
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal SpeI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal SpeI site found at 30
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal SpeI site found at 30
  • 1000
    COMPATIBLE 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

Fig.1 Design for sgRNA part, the sgRNA is preceded by a constant promoter, J23119.

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