pMinDis.X513-iNicSnFR3b-(V7)-ER
(Plasmid
#125123)
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PurposeDevelop a family of genetically encoded fluorescent biosensors for nicotine, termed iNicSnFRs thus enabling optical subcellular pharmacokinetics for nicotine
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 125123 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepMinDis.X513
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Backbone manufacturerJanelia
- Backbone size w/o insert (bp) 5005
- Total vector size (bp) 6655
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Vector typeMammalian Expression
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Selectable markersNA
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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Growth instructionsUse BL21(DE3) for bacterial expression and grow at 30 degrees C
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameiNicSnFR3a-(V7)-ER
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Alt nameiNicSnFR3a-(V7)-ER
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SpeciesMammalian
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Insert Size (bp)1650
- Promoter T7
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Tag
/ Fusion Protein
- Fusion protein -Endoplasmic retention motif (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BglII (unknown if destroyed)
- 3′ cloning site pstI (unknown if destroyed)
- 5′ sequencing primer T7
- 3′ sequencing primer BGH reverse (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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
Please note that a 6bp insertion and mutation V183A was identified in the NeoR/KanR cassette during Addgene's quality control process. We recommend testing NeoR selection before performing experiments. Vector was received from Janelia HHMI.
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:
pMinDis.X513-iNicSnFR3b-(V7)-ER was a gift from Henry Lester (Addgene plasmid # 125123 ; http://n2t.net/addgene:125123 ; RRID:Addgene_125123) -
For your References section:
Determining the pharmacokinetics of nicotinic drugs in the endoplasmic reticulum using biosensors. Shivange AV, Borden PM, Muthusamy AK, Nichols AL, Bera K, Bao H, Bishara I, Jeon J, Mulcahy MJ, Cohen B, O'Riordan SL, Kim C, Dougherty DA, Chapman ER, Marvin JS, Looger LL, Lester HA. J Gen Physiol. 2019 Feb 4. pii: jgp.201812201. doi: 10.1085/jgp.201812201. 10.1085/jgp.201812201 PubMed 30718376