pCrU6.4-SaCas9 cloning/aphVIII (pPH339)
From Andre Greiner, Peter Hegemann lab, Humboldt University-Berlin October 2017
Cloning vector for Staphylococcus aureus Cas9 (SaCas9) guide RNA transcription, controlled by the U6 snRNA promoter #4 (3’ of Cre15.g640800). The 20 bp target site can be inserted in a cut-ligation reaction as annealed oligos into an Esp3I cloning site following the protocol of Ann Ran (Ran et al. 2013). The immediate 4-bp sequence upstream of the transcriptional start site was changed to ACTT in all U6 constructs to simplify the cloning procedures.
Vector contains an aphVIII cassette for selection on paromomycin.