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PurposeTIR1-3FLAG fusion driven by a TUB1 promoter with a CAT drug selectable marker. The TIR1 CDS from Oryza sativa was codon optimized for T. gondii expression.
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Depositing Lab
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Depositing OrganizationWashington University in Saint Louis
Located in the United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 87258 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 87258-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 87258-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepBluescript SK(+)
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Backbone manufacturerStratagene
- Backbone size w/o insert (bp) 2925
- Total vector size (bp) 9655
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Vector typeToxoplasma expression
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Selectable markersCAT
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 1
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Gene/Insert nameTIR1
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SpeciesSynthetic; Oryza sativa
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Insert Size (bp)1806
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MutationCodon optimized for Toxoplasma gondii expression
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GenBank ID4335696
- Promoter TgTUB1 2717 bp
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Tag
/ Fusion Protein
- 3FLAG (C terminal on insert)
Cloning Information for Gene/Insert 1
- Cloning method Gibson Cloning
- 5′ sequencing primer cttgtgtgaagttcttgcgg
- 3′ sequencing primer gataaaggacacaggggtct
- (Common Sequencing Primers)
Gene/Insert 2
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Gene/Insert nameCAT
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Alt nameChloramphenicol acetyltransferase
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Alt nameCmR
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SpeciesSynthetic
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Insert Size (bp)663
- Promoter TgSAG1 361 bp
Cloning Information for Gene/Insert 2
- Cloning method Unknown
- 5′ sequencing primer cggttgtatgtcggtttcgc
- 3′ sequencing primer tgtgtattgacccatgtggc
- (Common Sequencing Primers)
Resource Information
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.
DNA (Catalog # 87258-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 # 87258-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:
pTUB1:OsTIR1-3FLAG, SAG1:CAT was a gift from David Sibley (Addgene plasmid # 87258 ; http://n2t.net/addgene:87258 ; RRID:Addgene_87258) -
For your References section:
Plasma Membrane Association by N-Acylation Governs PKG Function in Toxoplasma gondii. Brown KM, Long S, Sibley LD. MBio. 2017 May 2;8(3). pii: e00375-17. doi: 10.1128/mBio.00375-17. 10.1128/mBio.00375-17 PubMed 28465425