pRV-TdTom-MCS
(Plasmid #193711)

• Purpose: (Empty Backbone) Retroviral plasmid DNA for subcloning an oligonucleotide library encoding RNA barcodes
• Depositing Lab: Colin Akerman
• Publication: Plasmid for generating retroviral RNA barcodes (unpublished)

Backbone

• Vector backbone: New vector backbone generated from CAG-GFP (Addgene #16664)
• Backbone manufacturer: Fred Gage
• Backbone size (bp): 7426
• Modifications to backbone: We replaced GFP with TdTomato, using available AgeI/PmeI sites. We then inserted a multiple cloning site (MCS) downstream of TdTomato (after the stop codon and before the WPRE sequence) using annealed oligos.
• Vector type: Mammalian Expression, Retroviral
• Promoter: CAG

Growth in Bacteria

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

Cloning Information

• Cloning method: Restriction Enzyme

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This vector was derived from CAG-GFP (Addgene # 16664) - see author's map for more information. Expression is driven by the compound promoter CAG. This vector also contains the woodchuck hepatitis virus posttranscriptional element, which reportedly increases protein production by stabilizing the transcript.