pUltra-H2B-mTagBFP2
(Plasmid #161791)

• Purpose: 3rd generation lentiviral multicistronic vector for mammalian expression of EGFP, H2B-mTagBFP2, and one gene of interest
• Depositing Lab: Jorge Luis Arias-Arias
• Publication: pUltra series (unpublished)

Backbone

• Vector backbone: pUltra (Addgene Plasmid #24129)
• Backbone manufacturer: Malcolm Moore
• Backbone size w/o insert (bp): 8326
• Total vector size (bp): 9421
• Vector type: Mammalian Expression, Lentiviral ; Multicistronic

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: H2B-mTagBFP2
• Species: H. sapiens (human)
• Insert Size (bp): 1095
• Mutation: I174A in the mTagBFP2 protein according to PLoS ONE (2011) 6: e28674. doi:10.1371/journal.pone.0028674
• GenBank ID:
• Promoter: UbC
• Tags / Fusion Proteins:
  • mTagBFP2 (C terminal on insert)
  • EGFP (N terminal on backbone)

Cloning Information

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

Resource Information

• Supplemental Documents:
  • Live-cell imaging of pUltra-mTagBFP2 in BHK-21 cells

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

pUltra-H2B-mTagBFP2 was cloned by Jorge L. Arias-Arias at Universidad de Costa Rica ()

This vector allows the multicistronic expression of EGFP, H2B-mTagBFP2, and one gene of interest in mammalian cells. The construct contains two 2A peptides coding sequences and two cloning positions flanked by restriction sites:

Position 1 (5' XbaI – 3' BamHI): Occupied by the H2B-mTagBFP2 insert.

Position 2 (5' NheI – 3' EcoRI or SaII): Available for cloning one gene of interest.

The vector is suitable for transient transfection, but could also be used to produce lentiviral particles for stable expression in mammalian cells. For lentivirus packaging use the additional plasmids psPAX2 (Addgene Plasmid #12260) and pMD2.G (Addgene Plasmid #12259).