PX552
(Plasmid #60958)

• Purpose: pAAV-U6sgRNA(SapI)_hSyn-GFP-KASH-bGH (SpGuide acceptor). AAV plasmid for sgRNA cloning. GFP-KASH fusion facilitates FACS sorting of cells and nuclei.
• Depositing Lab: Feng Zhang
• Publication: Swiech et al Nat Biotechnol. 2014 Oct 19. doi: 10.1038/nbt.3055.

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Backbone

• Vector backbone: pAAV
• Backbone size w/o insert (bp): 2905
• Total vector size (bp): 5314
• Vector type: Mammalian Expression, Mouse Targeting, AAV, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: U6_(SpaI)_sgRNA
• Species: Synthetic
• Insert Size (bp): 351
• Promoter: U6

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: cgcgtgagggcctatttcc
• 3′ sequencing primer: gcaagtgggttttaggaccag

Gene/Insert 2

• Gene/Insert name: Syn_EGFP-KASH
• Species: Synthetic
• Insert Size (bp): 349
• Promoter: Synapsin

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ctgcgtatgagtgcaagtgggtt
• 3′ sequencing primer: ccgttggtgtaccgcagcatg

Resource Information

• Supplemental Documents:
  • PX552
  • protocol for cloning sgRNA
• Articles Citing this Plasmid:
  • 54 References

Terms and Licenses

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