pC0049-EF1a-dPSPCas13b-NES-HIV, H133A/H1058A
(Plasmid #103865)

• Purpose: Catalytically inactive PSPCas13b. RNase activity is inactivated by mutation of catalytic histidine residues in the HEPN domains (H133A/H1058A).
• Depositing Lab: Feng Zhang
• Publication: Cox et al Science. 2017 Oct 25. pii: eaaq0180. doi: 10.1126/science.aaq0180.

Backbone

• Vector backbone: Unknown
• Backbone size w/o insert (bp): 9360
• Total vector size (bp): 12606
• Vector type: Mammalian Expression, Lentiviral, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: PspCas13b
• Species: Prevotella sp. P5-125
• Insert Size (bp): 3270
• Mutation: Contains the H133A and H1058A mutations which inactivate the HEPN RNase domains of Cas13b.
• GenBank ID: WP_044065294
• Promoter: EF1a
• Tags / Fusion Proteins:
  • HIV NES (C terminal on insert)
  • 3xHA (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: EF1a-F

Resource Information

• Supplemental Documents:
  • Annotated GenBank file
• Addgene Notes:
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor Comments

Benchling link: https://benchling.com/s/seq-gF8829PyBC9oBkazyblL