pCT
(Plasmid #60620)

• Purpose: Plasmid encoding iCas9 and tracrRNA on pRS415 backbone
• Depositing Lab: Huimin Zhao
• Publication: Bao et al ACS Synth Biol. 2014 Sep 19.

Backbone

• Vector backbone: pRS415
• Backbone size w/o insert (bp): 6000
• Total vector size (bp): 10956
• Modifications to backbone: All the BsmBI sites were removed
• Vector type: Yeast Expression, CRISPR
• Selectable markers: LEU2

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: iCas9
• Species: Streptococcus pyogenes
• Insert Size (bp): 4179
• Mutation: changed Aspartate 147 to Tyrosine, Proline 411 to Threonine
• Promoter: TEF1p
• Tags / Fusion Proteins:
  • FLAG (N terminal on insert)
  • SV40 NLS (N terminal on insert)
  • SV40 NLS (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: Unknown
• 3′ sequencing primer: Unknown

Gene/Insert 2

• Gene/Insert name: tracrRNA
• Species: Streptococcus pyogenes
• Insert Size (bp): 86
• Promoter: RPR1p

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: DraIII (unknown if destroyed)
• 3′ cloning site: NcoI (unknown if destroyed)
• 5′ sequencing primer: Unknown
• 3′ sequencing primer: Unknown

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Dr. Feng Zhang, Dr. Jennifer A. Doudna
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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