Difference between revisions of "Part:BBa K1699005:Design"

(→‎References)
(→‎References)
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1. In vivo genome editing using Staphylococcus aureus Cas9.  
 
1. In vivo genome editing using Staphylococcus aureus Cas9.  
 
<br />http://10.1038/nature14299  
 
<br />http://10.1038/nature14299  
 +
 
2. Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas
 
2. Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas
 
<br />http://pubs.acs.org/doi/full/10.1021/sb400081r
 
<br />http://pubs.acs.org/doi/full/10.1021/sb400081r

Revision as of 20:01, 5 September 2015

gRNA for dCas9-VP64 targeting synthetic activation promoter pMLPm under U6 promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 250
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

gRNA in SaCas9 scaffold under U6 promoter. Designed to complement 3 sites on MLP synthetic promoter.
Cloned into pSBC13 using EcoRI and PstI restriction sites.
F. tagtcatggcggccgcgtcgacG
R. actgacatgcggccgcttaattaaC

Source

gMLP from IDT de-novo synthesis. Cloning with U6 to pSBC13 was done by team BGU 2015.

References

1. In vivo genome editing using Staphylococcus aureus Cas9.
http://10.1038/nature14299

2. Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas
http://pubs.acs.org/doi/full/10.1021/sb400081r