pAAV2-UbC-3xNLS-mScarlet-I
(Plasmid #191099)

• Purpose: To express a bright monomeric red FP to label eucaryotic cell nuclei. To be used in tissue clearing methods and other fluorescent microscopy methods
• Depositing Labs: Murray Blackmore, Pantelis Tsoulfas
• Publication: Wang et al Elife. 2022 Jul 15;11. pii: 76254. doi: 10.7554/eLife.76254.

Backbone

• Vector backbone: pAAV-SIBR-anti_I
• Backbone manufacturer: Chung K-H et al., Polycistronic RNA polymerase II expression vectors for RNA interference based on BIC/miR-155. NAR 2006 vol 24
• Backbone size w/o insert (bp): 5322
• Total vector size (bp): 6130
• Modifications to backbone: The anti-luciferase cassette was deleted
• Vector type: Mammalian Expression, AAV, Synthetic Biology

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

• Gene/Insert name: 3xNLS-mScarlet-I
• Species: Synthetic
• Insert Size (bp): 798
• Mutation: optimized to human codon usage
• Promoter: UbC
• Tag / Fusion Protein:
  • Three repeats of the nuclear localizzation sequences from SV40 fused to mScarlet-I.FPbase ID: 6VVTK (N terminal on insert) (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI, NheI (not destroyed)
• 3′ cloning site: None (destroyed during cloning)
• 5′ sequencing primer: pCAF-F
• 3′ sequencing primer: hGH-PA-R

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry