pCRISPR-S12
(Plasmid #84031)

• Purpose: To episomally express codon optimized Cas9 and chimeric guide RNA
• Depositing Lab: Sen Wu
• Publication: Li et al Protein Cell. 2016 Sep;7(9):689-91. doi: 10.1007/s13238-016-0299-9.

Backbone

• Vector backbone: pCEP4
• Backbone manufacturer: Invitrogen
• Vector type: Mammalian Expression
• Selectable markers: Puromycin, Hygromycin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: hSpCas9
• Promoter: CMV
• Tag / Fusion Protein:
  • 3x FLAG (N terminal on insert)

Cloning Information for Gene/Insert 1

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

Gene/Insert 2

• Gene/Insert name: eGFP
• Species: jellyfish
• Promoter: CMV (downstream of F2A self-cleaving peptide)

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: unknown
• 3′ sequencing primer: BGH-rev

Terms and Licenses

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

Depositor Comments

Please note for gRNA cloning: gRNA sequence should be constructed separately in a different u6 containing vector such as pX330, and subcloned into BamHI/NotI sites of pCRISPR-S12.