Coding

Part:BBa_K1699005:Design

Designed by: Emil Ruvinov   Group: iGEM15_BGU_Israel   (2015-08-09)

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

Designed using Benchling to meet registry standards. Synthesized by IDT. Cloned into pSB1C3 using EcoRI and PstI restriction sites.

Source

Sequence of gRNA:

Tunable and multifunctional eukaryotic transcription factors based on CRISPR/Cas. Farzadfard F, Perli SD, Lu TK. ACS Synth Biol. 2013 Oct 18;2(10):604-13. doi: 10.1021/sb400081r. Epub 2013 Sep 11. http://www.ncbi.nlm.nih.gov/pubmed/23977949

Sequence of U6 promoter:

In vivo genome editing using Staphylococcus aureus Cas9. Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F. Nature. 2015 Apr 9;520(7546):186-91. doi: 10.1038/nature14299. Epub 2015 Apr 1. http://www.ncbi.nlm.nih.gov/pubmed/25830891