pUltra-Plus-H2B-EGFP
(Plasmid #161776)

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• Publication: pUltra series (unpublished)

Backbone

• Vector backbone: pUltra-Plus (Addgene Plasmid #161775)
• Backbone manufacturer: Jorge L. Arias-Arias
• Backbone size w/o insert (bp): 7611
• Total vector size (bp): 8724
• Modifications to backbone: Destruction of BamHI restriction site
• Vector type: Mammalian Expression, Lentiviral ; Bicistronic

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-EGFP from Addgene Plasmid #26790
• Species: H. sapiens (human)
• Insert Size (bp): 1113
• GenBank ID:
• Promoter: UbC
• Tag / Fusion Protein:
  • EGFP (C terminal on insert)

Cloning Information

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

Resource Information

• Supplemental Documents:
  • Live-cell imaging of pUltra-Plus-H2B-EGFP in BHK-21 cells

Terms and Licenses

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

Depositor Comments

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

This vector allows bicistronic expression in mammalian cells. The construct contains two 2A peptides coding sequences and three cloning positions flanked by restriction sites:

Position 1 (5' AgeI – 3' EcoRV): Not available for cloning as the H2B-EGFP insert contains another AgeI restriction site. This extra AgeI restriction site could be removed by site-directed mutagenesis in order to rescue this cloning position and obtain a multicistronic construct for the expression of the H2B-EGFP insert and two additional genes of interest.

Position 2 (5' XbaI – 3' BamHI/BgIII destroyed during cloning): Occupied by the H2B-EGFP insert.

Position 3 (5' NheI – 3' 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).