nls-NgAgo-GK
(Plasmid #78253)

• Purpose: Mammalian expression of NgAgo (Argonaute from N. gregoryi).
• Depositing Lab: Chunyu Han
• Publication: Gao et al Nat Biotechnol. 2016 May 2. doi: 10.1038/nbt.3547.

Backbone

• Vector backbone: pEGFP-N1
• Backbone size w/o insert (bp): 4000
• Total vector size (bp): 6679
• Modifications to backbone: pEGFP-N1 without the eGFP
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: NgAgo
• Alt name: Argonaute
• Species: Natronobacterium gregoryi
• Insert Size (bp): 2712
• GenBank ID: AFZ73749.1
• Promoter: CMV
• Tag / Fusion Protein:
  • a nuclear localization signal (NLS) (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: SV40pA-R

Resource Information

• Supplemental Documents:
  • Annotated sequence
  • General Protocol for NgAgo-mediated genome editing
• Addgene Notes:
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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