pRSET a sfGCaMP6s-T78H
(Plasmid
#100023)
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PurposeBacterial expression of fluorescent reporter for calcium signaling, based off of GCaMP6s. cpEGFP replaced with a superfolder cpGFP variant containing T78H mutation
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 100023 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepRSET A
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Backbone manufacturerThermo Fisher Scientific
- Backbone size w/o insert (bp) 2973
- Total vector size (bp) 4218
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Vector typeBacterial Expression
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namesfGCaMP6s-T78H
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Insert Size (bp)1245
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MutationcpEGFP replaced with superfolder cpGFP and T78H mutation reintroduced
- Promoter T7
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Tags
/ Fusion Proteins
- 6x HIS tag (N terminal on backbone)
- Xpress_EK tag (N terminal on backbone)
Cloning Information
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer T7
- 3′ sequencing primer T7 term (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made byThe M13 peptide and CaM from GCaMP6s (derived from Addgene plasmid #40753 - Douglas Kim) were used in the construction of this plasmid.
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
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.
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For your Materials & Methods section:
pRSET a sfGCaMP6s-T78H was a gift from Wolf Frommer (Addgene plasmid # 100023 ; http://n2t.net/addgene:100023 ; RRID:Addgene_100023) -
For your References section:
Ratiometric Matryoshka biosensors from a nested cassette of green- and orange-emitting fluorescent proteins. Ast C, Foret J, Oltrogge LM, De Michele R, Kleist TJ, Ho CH, Frommer WB. Nat Commun. 2017 Sep 5;8(1):431. doi: 10.1038/s41467-017-00400-2. 10.1038/s41467-017-00400-2 [pii] PubMed 28874729