ROSA26(MultiFPsΔPuro)
(Plasmid #140759)

• Purpose: Targeting vector to Gt(ROSA)26 locus for conditional expression of distinct FPs (Venus, mCherry, and mCerulean) responding to each site-specific-recombinase activity (Cre, Dre, and phiC31o).
• Depositing Lab: Hideyuki Okano
• Publication: Serizawa et al Development. 2019 Nov 4;146(21). pii: dev.174938. doi: 10.1242/dev.174938.

Backbone

• Vector backbone: pCAGGS
• Backbone size w/o insert (bp): 2396
• Total vector size (bp): 18841
• Vector type: Mouse Targeting
• Selectable markers: Puromycin

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: puromycin-N-acetyltransferase
• Alt name: Pac
• Species: Synthetic
• Insert Size (bp): 600
• Promoter: CAGGS

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: XbaI (not destroyed)
• 3′ cloning site: SpeI (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: n/a

Gene/Insert 2

• Gene/Insert name: Venus
• Alt name: YFP
• Alt name: GFP variant
• Species: Synthetic
• Insert Size (bp): 720
• Promoter: CAGGS

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: SpeI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: n/a

Gene/Insert 3

• Gene/Insert name: mCherry
• Alt name: RFP variant
• Species: Synthetic
• Insert Size (bp): 708
• Promoter: CAGGS

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: n/a

Gene/Insert 4

• Gene/Insert name: mCerulean
• Alt name: CFP
• Alt name: GFP variant
• Species: Synthetic
• Insert Size (bp): 720
• Promoter: CAGGS

Cloning Information for Gene/Insert 4

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: n/a

Resource Information

• Supplemental Documents:
  • ROSA26(MultiFPsΔPuro).gb
• A portion of this plasmid was derived from a plasmid made by: Venus was cloned from pTP1-Venus (Kohyama et al., Dev. Biol. 286, 311-325, 2005); mCherry was cloned from pG-PB-Zscan4c-mCherry-polyAfloxPGKneo, a gift from Minoru Ko, Keio University School of Medicine, Japan; mCerulean was cloned from Cerulean (Addgene plasmid #15214, Rizzo et al., Nat. Biotechnol. 22, 445-449, 2004), a gift from Dave Piston, Washington University School of Medicine, USA.

Terms and Licenses

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