pRSET-RCaMP1h
(Plasmid #42874)

• Purpose: a single-wavelength red-shifted genetically encoded calcium indicator constructed from calmodulin and cp-mRuby
• Depositing Lab: Loren Looger
• Publication: Akerboom et al Front Mol Neurosci. 2013;6:2. doi: 10.3389/fnmol.2013.00002. Epub 2013 Mar 4.

Backbone

• Vector backbone: pRSETA
• Backbone manufacturer: Life Technologies (Invitrogen)
• Backbone size w/o insert (bp): 2753
• Total vector size (bp): 4082
• Modifications to backbone: No modifications to backbone are present besides cloning in Multiple Cloning Site.
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Red Fluorescent Calcium binding Protein
• Alt name: RCaMP1.035
• Alt name: RCaMP181
• Species: R. norvegicus (rat), Synthetic
• Insert Size (bp): 1329
• Promoter: T7
• Tags / Fusion Proteins:
  • Xpress™ epitope (N terminal on insert)
  • EK (Enterokinase) Recognition Site (N terminal on insert)
  • His Tag (6x) (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: TAATACGACTCACTATAGG
• 3′ sequencing primer: GCTAGTTATTGCTCAGCG

Resource Information

• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

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

Depositor Comments

Gene was cloned using BaMHI and HindIII - But part of the vector backbone then becomes part of the protein (leader sequence containing histag) - using NdeI and HindIII will completely remove the open reading frame/gene.