Lenti-sgRNA(MS2)-puro-barcode
(Plasmid #89493)

• Purpose: Cloning backbone for sgRNA, also has a barcode that allows detection from scRNA-seq
• Depositing Lab: Gary Hon
• Publication: Xie et al Mol Cell. 2017 Apr 20;66(2):285-299.e5. doi: 10.1016/j.molcel.2017.03.007. Epub 2017 Apr 13.

Backbone

• Vector backbone: plenti
• Backbone size w/o insert (bp): 10000
• Total vector size (bp): 8401
• Modifications to backbone: The sgRNA and Cas9 gene were replaced with sgRNA(MS2) backbone and an barcode insertion region was introduced between WPRE and 3'LTR.
• Vector type: Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Growth instructions: Only amplify in recombinase deficient bacteria (eg. Stbl3 or HST08).
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: sgRNA (with MS2 loop)
• Alt name: lentiGuide Barcode
• gRNA/shRNA sequence: empty backbone
• Species: Synthetic
• Promoter: U6

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ATCTTGTGGAAAGGACGAAACACCGGAGACGGGATACC
• 3′ sequencing primer: ctcaagatctagttacgccaagcttAAAAAAgcaccgact

Resource Information

• Supplemental Documents:
  • Annotated GenBank file

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry