pRRLsin-SV40 T antigen-IRES-mCherry
(Plasmid #58993)

• Purpose: lentiviral expression of SV40 large T antigen and mCherry red fluorescent reporter
• Depositing Lab: Snorri Thorgeirsson
• Publication: Holczbauer et al Gastroenterology. 2013 Jul;145(1):221-31. doi: 10.1053/j.gastro.2013.03.013. Epub 2013 Mar 19.

Backbone

• Vector backbone: pRRLsin.cPPT.PGK/GFP.WPRE
• Backbone size w/o insert (bp): 6110
• Total vector size (bp): 11405
• Modifications to backbone: NotI site at nt 924 was eliminated
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: mCherry red fluorescence

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: CMV enhancer-chicken beta actin promoter
• Insert Size (bp): 1730
• Mutation: EcoRV site in MCS converted to BamHI site

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (destroyed during cloning)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: none
• 3′ sequencing primer: pCAG-F

Gene/Insert 2

• Gene/Insert name: SV40 large T antigen
• Alt name: SV40gp6
• Insert Size (bp): 2200
• Promoter: CMV enhancer-chicken beta actin

Cloning Information for Gene/Insert 2

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

Gene/Insert 3

• Gene/Insert name: IRES-mCherry red fluorescent protein
• Species: Synthetic
• Insert Size (bp): 1360
• Promoter: CMV enhancer-chicken beta actin

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: IRES-R
• 3′ sequencing primer: mCherry-F

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The mCherry sequence was obtained from Dr. Roger Tsien. Other genes were obtained from plasmids purchased from Addgene
• Articles Citing this Plasmid:
  • 6 References

Terms and Licenses

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