LV-indLS2
(Plasmid #123068)

• Purpose: Lentiviral construct that delivers a tamoxifen inducible Cre. Upon recombination, luc2 and mStrawberry express. Used to image spontaneous tumorigenesis or subclonal cell populations in vivo.
• Depositing Lab: Kevin Brindle
• Publication: Rodriguez et al J Pathol. 2014 Mar;232(4):449-57. doi: 10.1002/path.4313.

Backbone

• Vector backbone: pBOB
• Backbone manufacturer: Verma lab (Salk Institute)
• Backbone size w/o insert (bp): 7800
• Total vector size (bp): 14358
• Vector type: Mammalian Expression, Lentiviral, Luciferase ; Fluorescence
• Selectable markers: luciferase or mStrawberry expression following Cre mediated recombination

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: CreERT2 with intron
• Species: Synthetic; Bacteriophage P1
• Insert Size (bp): 2127
• Mutation: synthetic intron added as indicated
• Promoter: CAGGS (flanked by inverted lox66 and lox71 sequences)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: ggcgttactatgggaaca

Gene/Insert 2

• Gene/Insert name: Firefly luciferase
• Alt name: Luc2
• Species: Photinus pyralis
• Insert Size (bp): 1659
• Promoter: CAGGS (after Cre mediated inversion)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: acattacctacgccgagt

Gene/Insert 3

• Gene/Insert name: mStrawberry
• Species: Discosoma sp.
• Insert Size (bp): 738
• Promoter: CAGGS (after Cre mediated inversion)

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: TGATGAACTTCGAGGACG

Resource Information

• Supplemental Documents:
  • LV-indLS2.gbk
  • List and position of key plasmid sequence features

Terms and Licenses

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