MSCV-Cas9-2A-BFP
(Plasmid #164662)

• Purpose: Retroviral introduction of Cas9 into mammalian cell line coupled to expression of blue fluorescent protein
• Depositing Lab: Daniel Hodson
• Publication: Caeser et al Nat Commun. 2019 Oct 4;10(1):4543. doi: 10.1038/s41467-019-12494-x.

Backbone

• Vector backbone: MSCV_Cas9_puro
• Backbone manufacturer: Addgene 65655
• Backbone size w/o insert (bp): 10475
• Total vector size (bp): 10071
• Modifications to backbone: PGK promoter and PuroR removed, T2A tagBFP inserted
• Vector type: Mammalian Expression, Retroviral, CRISPR

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: Cas9
• Alt name: S. pyogenes CRISPR-Cas9
• Species: Streptococcus pyogenes
• Insert Size (bp): 4101
• Promoter: MSSV_LTR

Cloning Information for Gene/Insert 1

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CTTTATCCAGCCCTCAC
• 3′ sequencing primer: AGCGAGTCAGTGAGCGAG

Gene/Insert 2

• Gene/Insert name: BFP
• Alt name: blue fluorescent protein
• Alt name: TagBFP
• Species: Synthetic
• Insert Size (bp): 702
• Promoter: MSSV_LTR

Cloning Information for Gene/Insert 2

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CTTTATCCAGCCCTCAC
• 3′ sequencing primer: AGCGAGTCAGTGAGCGAG

Resource Information

• Supplemental Documents:
  • 164662 Genbank file.gb
• A portion of this plasmid was derived from a plasmid made by: Addgene vector 65655

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry