pcDNA5-H2B_Halo_T2A_EGFP
(Plasmid #135444)

• Purpose: Mamalian cell expression of nuclear HaloTag7 and cytosolic EGFP
• Depositing Lab: Kai Johnsson
• Publication: Frei et al Nat Commun. 2019 Oct 8;10(1):4580. doi: 10.1038/s41467-019-12480-3.

Backbone

• Vector backbone: pcDNA5/FRT/TO
• Backbone manufacturer: thermofisher
• Backbone size w/o insert (bp): 5137
• Total vector size (bp): 7195
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: H2B-HaloTag7 and EGFP
• Insert Size (bp): 2061
• Promoter: CMV/TetO
• Tag / Fusion Protein:
  • H2B (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV_Fwd
• 3′ sequencing primer: BGH_Rev

Resource Information

• Supplemental Documents:
  • pcDNA5-H2B_Halo_T2A_EGFP.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • HaloTag Vector Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Please note this plasmid appears to be unstable during growth in bacteria. We recommend screening 10 smaller colonies after streaking from the original stab. It may be necessary to cut the plasmid with a single-cutting enzyme, gel purify the high molecular weight band, and religate to restore the plasmid.