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PurposeExpresses a DHFR-mCherry fusion protein and uses for a CRISPR gene deletion
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Depositing Lab
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Depositing OrganizationWashington University in Saint Louis
Located in United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 70147 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 70147-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 70147-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepUC19
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Backbone manufacturerinvitrogen
- Backbone size w/o insert (bp) 2100
- Total vector size (bp) 7317
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Modifications to backboneNO
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Vector typeExpression in Toxoplasma gondii
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Selectable markersDHFR
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 namea selectable dihydrofolate reductase-thymidylate synthase marker
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Alt nameDHFR-TS-mCherry
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SpeciesToxoplasma gondii
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Insert Size (bp)4586
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GenBank IDXM_002367211.1 XM_002367211.1
- Promoter DHFR 5' UTR
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Tag
/ Fusion Protein
- mCherry (C terminal on insert)
Cloning Information
- Cloning method Unknown
- 5′ sequencing primer M13F
- 3′ sequencing primer M13R
- (Common Sequencing Primers)
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
Kevin Brown and Shaojun Long jointly built the plasmid in David Sibley lab, for CRISPR genetic operations.
DNA (Catalog # 70147-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 70147-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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:
pLoxP-DHFR-mCherry was a gift from David Sibley (Addgene plasmid # 70147 ; http://n2t.net/addgene:70147 ; RRID:Addgene_70147) -
For your References section:
Analysis of non-canonical calcium dependent protein kinases in Toxoplasma gondii by targeted gene deletion using CRISPR/Cas9. Long S, Wang Q, Sibley LD. Infect Immun. 2016 Jan 11. doi: 10.1128/IAI.01173-15 10.1128/IAI.01173-15 PubMed 26755159
Map uploaded by the depositor.
