3xFLAG-dCas9/MSCV-EGFP
(Plasmid #82613)

• Purpose: Expresses 3xFLAG-dCas9 in mammalian cells for enChIP analysis to purify specific genomic regions of interest.
• Depositing Lab: Hodaka Fujii
• Publication: Fujita et al Genes Cells. 2018 Feb 26. doi: 10.1111/gtc.12573.

Backbone

• Vector backbone: MSCV-EGFP
• Backbone size w/o insert (bp): 6421
• Total vector size (bp): 10822
• Vector type: Retroviral
• Selectable markers: EGFP

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: 3xFLAG-dCas9
• Species: Synthetic
• Insert Size (bp): 4212
• Mutation: human codon-optimized, D10A + H840A
• Promoter: LTR
• Tags / Fusion Proteins:
  • 3xFLAG tag (N terminal on insert)
  • NLS (nuclear localization signal) (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Hpa I (destroyed during cloning)
• 3′ cloning site: Hap I (destroyed during cloning)
• 5′ sequencing primer: MSCV (5'-CCCTTGAACCTCCTCGTTCGACC-3')
• 3′ sequencing primer: EGFP-N (5'-CGTCGCCGTCCAGCTCGACCAG-3)

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

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