Coding

Part:BBa_K1699005:Design

Designed by: Emil Ruvinov   Group: iGEM15_BGU_Israel   (2015-08-09)
Revision as of 19:59, 5 September 2015 by Shalevgo (Talk | contribs) (→‎Design Notes)

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

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