pcDNA-NLS-ZapCmR2
(Plasmid
#59012)
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PurposeNuclear zinc-binding FRET biosensor
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Depositing Lab
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Sequence Information
Full plasmid sequence is not available for this item.
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 59012 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA3.1(+)
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5500
- Total vector size (bp) 7200
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Modifications to backboneSV40 NLS sequence added upstream of BamHI site
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Vector typeMammalian Expression
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Selectable markersNeomycin (select with G418)
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)DH10B
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameNLS-ZapCmR2 genetically encoded Zn(II) sensor
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SpeciesSynthetic
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Insert Size (bp)1686
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MutationSee comment
- Promoter CMV
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Tag
/ Fusion Protein
- SV40 NLS (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer T7 Forward
- 3′ sequencing primer BGH Reverse (Common Sequencing Primers)
Resource Information
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Article Citing 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.
Depositor Comments
Genetically encoded, ratiometric, fluorescent biosensor. Contains NLS sequence at N-terminus. Zn(II)-binding domain derived from the first 2 zinc fingers of Zap 1 from Saccharomyces cerevisiae. Contains Clover and mRuby fluorescent proteins.
The Zn(II) binding domain of ZapCmR2 is derived from ZapCmR1.1. The first cysteine in the second zinc finger is mutated to histidine.
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:
pcDNA-NLS-ZapCmR2 was a gift from Amy Palmer (Addgene plasmid # 59012 ; http://n2t.net/addgene:59012 ; RRID:Addgene_59012) -
For your References section:
New alternately colored FRET sensors for simultaneous monitoring of Zn(2)(+) in multiple cellular locations. Miranda JG, Weaver AL, Qin Y, Park JG, Stoddard CI, Lin MZ, Palmer AE. PLoS One. 2012;7(11):e49371. doi: 10.1371/journal.pone.0049371. Epub 2012 Nov 16. 10.1371/journal.pone.0049371 PubMed 23173058
Map uploaded by the depositor.