pRH2502
(Plasmid #84379)

• Purpose: Expression of dcas9 D10A H840A from a TetR-regulated uvtetO promoter
• Depositing Lab: Robert Husson
• Publication: Singh et al Nucleic Acids Res. 2016 Jul 12. pii: gkw625.

Backbone

• Vector backbone: pTC-0X-1L
• Backbone manufacturer: Addgene 20315
• Backbone size w/o insert (bp): 4640
• Total vector size (bp): 8779
• Modifications to backbone: The PacI site near the T4g32 sites was removed. PacI sites were introduced 15 bp after the NdeI site encompassing the start codon of lacZI, and 5’ of the SV40 PA terminator. lacZI was removed and dcas9 was inserted under the control of uv15tetO promoter at NdeI and PacI cloning sites
• Vector type: Bacterial Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: dcas9
• Alt name: cas9 D10A and H840A
• Species: Streptococcus pyogenes
• Insert Size (bp): 4139
• Mutation: Aspartate at position 10 changed to Alanine, Histidine at position 840 changed to Alanine
• GenBank ID: AAK33936.1
• Promoter: uv15tetO

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: PacI (not destroyed)
• 5′ sequencing primer: N/A
• 3′ sequencing primer: N/A

Resource Information

• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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