hsp70l-loxP-mCherry-STOP-loxP-H2B-GFP_cryaa-cerulean
(Plasmid #24334)

• Depositing Lab: Didier Stainier
• Publication: Hesselson et al Proc Natl Acad Sci U S A. 2009 Sep 1. 106(35):14896-901.

Backbone

• Vector backbone: pBluescript
• Backbone size w/o insert (bp): 3000
• Vector type: Worm Expression, Cre/Lox

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: H2B
• Species: D. rerio (zebrafish)
• Mutation: mCherry driven by an HSP70 promoterCerulean driven by a CrystalineA promoter*
• Tag / Fusion Protein:
  • GFP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: SV40pA

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

This plasmid was constructed using 1.5 kb of the hsp70l promoter. The floxed mCherry-STOP and H2b_GFP fragments were PCR amplified individually and sequentially cloned downstream of the hsp70l promoter. The mCherry-STOP cassette serves as a heat-shock induction marker prevents read-through translation of the H2B-GFP fusion protein, which is only expressed in cells that have undergone a Cre mediated excision event. cryaa-Cerulean was inserted downstream of HSB-GFP in the reverse orientation.

*H2B has D26G and V119I mutations. According to the depositor, GFP expression from this construct was always nuclear so there is no reason to suspect that the mutations affect the function (nuclear localization) of the H2B fragment.