Purposea single-wavelength red-shifted genetically encoded calcium indicator constructed from calmodulin and cp-mRuby
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||42874||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Backbone manufacturerLife Technologies (Invitrogen)
- Backbone size w/o insert (bp) 2753
- Total vector size (bp) 4082
Modifications to backboneNo modifications to backbone are present besides cloning in Multiple Cloning Site.
Vector typeBacterial Expression
Growth in Bacteria
Growth Strain(s)XL1 Blue
Copy numberHigh Copy
Gene/Insert nameRed Fluorescent Calcium binding Protein
SpeciesR. norvegicus (rat), Synthetic
Insert Size (bp)1329
- Promoter T7
/ Fusion Proteins
- Xpress™ epitope (N terminal on insert)
- EK (Enterokinase) Recognition Site (N terminal on insert)
- His Tag (6x) (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site NdeI (not destroyed)
- 3′ cloning site HindIII (not destroyed)
- 5′ sequencing primer TAATACGACTCACTATAGG
- 3′ sequencing primer GCTAGTTATTGCTCAGCG (Common Sequencing Primers)
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pRSET-RCaMP1h was a gift from Loren Looger (Addgene plasmid # 42874 ; http://n2t.net/addgene:42874 ; RRID:Addgene_42874)
For your References section:Genetically encoded calcium indicators for multi-color neural activity imaging and combination with optogenetics. Akerboom J, Carreras Calderon N, Tian L, Wabnig S, Prigge M, Tolo J, Gordus A, Orger MB, Severi KE, Macklin JJ, Patel R, Pulver SR, Wardill TJ, Fischer E, Schuler C, Chen TW, Sarkisyan KS, Marvin JS, Bargmann CI, Kim DS, Kugler S, Lagnado L, Hegemann P, Gottschalk A, Schreiter ER, Looger LL. Front Mol Neurosci. 2013;6:2. doi: 10.3389/fnmol.2013.00002. Epub 2013 Mar 4. 10.3389/fnmol.2013.00002 PubMed 23459413