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Depositing Lab
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Depositing OrganizationRockefeller University
Located in United States of America -
Publication
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Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 8709 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 8709-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 8709-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepRc/CMV
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5500
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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)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameStat3 Y705F
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Alt nameStat3
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SpeciesM. musculus (mouse)
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Insert Size (bp)2600
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MutationY705F
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GenBank IDU06922
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Entrez GeneStat3 (a.k.a. 1110034C02Rik, Aprf)
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Tag
/ Fusion Protein
- FLAG (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NotI (not destroyed)
- 3′ cloning site ApaI (not destroyed)
- 5′ sequencing primer T7
- 3′ sequencing primer SP6
- (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
Diagnostic Restriction Digest with BamHI (3.2kb, 2.8kb, and 2.0kb). Due to a stop codon error made when tagging the Stat3 clone with FLAG (DYKDDDDK) additional residues follow the tag (GPYSIVSPKC STOP). The ApaI cloning site is GGG CCC, which encodes the G and P residues coming after the FLAG tag. Therefore, if the insert is removed with a NotI and ApaI digestion, it will not have a stop codon included.
* E16K mutation found during QC
DNA (Catalog # 8709-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 # 8709-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:
Stat3 Y705F Flag pRc/CMV was a gift from Jim Darnell (Addgene plasmid # 8709 ; http://n2t.net/addgene:8709 ; RRID:Addgene_8709) -
For your References section:
Mapping of Stat3 serine phosphorylation to a single residue (727) and evidence that serine phosphorylation has no influence on DNA binding of Stat1 and Stat3. Wen Z, Darnell JE Jr. Nucleic Acids Res 1997 Jun 1;25(11):2062-7. 10.1093/nar/25.11.2062 PubMed 9153303
