pLenti.PGK.H2B-chFP.W
(Plasmid #51007)

• Purpose: Lentivector that expresses H2B-chFP from internal human PGK promoter
• Depositing Lab: Rusty Lansford
• Publication: Huss et al Development. 2015 Jul 24. pii: dev.121392.

Backbone

• Vector backbone: pRRLsin.cPPT.PGKp.eGFP.wpre
• Backbone size w/o insert (bp): 6686
• Total vector size (bp): 7793
• Modifications to backbone: pLenti.pgk:H2B-chFP lentivector was derived from the pRRLsin.cPPT.PGKp.eGFP.wpre lentivector that contains the constitutively expressed human phosphoglycerate kinase promoter (De Palma et al., 2003). The membrane localization tag of the pLenti.pgk:membrane-mCherry lentivector (derived from pRRLsin.cPPT.PGKp.eGFP.wpre) was removed by AgeI/BamHI and replaced by the AgeI/BglII Histone 2B region of pH2B-GFP (Kanda et al., 1998) to generate pLV.pgk:H2B-chFP.
• Vector type: Mammalian Expression, Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: H2B-ChFP
• Alt name: CherryFP
• Insert Size (bp): 1107
• Promoter: human PGK
• Tag / Fusion Protein:
  • histone2B (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Age1 (not destroyed)
• 3′ cloning site: BamH1 (destroyed during cloning)
• 5′ sequencing primer: T3
• 3′ sequencing primer: SV40pA-R

Resource Information

• Articles Citing this Plasmid:
  • 6 References

Terms and Licenses

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

Depositor Comments

Please note that the H2B sequence in this plasmid was derived from H2B-GFP (Addgene plasmid# 11680) and contains the same differences (D26G and V119I) compared to GenBank reference sequences, such as NP_066402.2.