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Purposefor plasmid integration using the Flp-In system. AID at N-terminal
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Depositing Lab
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Depositing OrganizationLudwig Institute For Cancer Research, San Diego Branch
Located in United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 47330 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 47330-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 47330-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepcDNA5/FRT/TO
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Backbone manufacturerInvitrogen
- Total vector size (bp) 9850
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Vector typeMammalian Expression
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Selectable markersHygromycin
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)XL10 Gold
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameBUB1 mitotic checkpoint serine/threonine kinase B
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Alt nameBub1B
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Alt nameBubR1
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SpeciesH. sapiens (human)
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Insert Size (bp)4590
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MutationsiRNA resistant
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GenBank IDNM_001211.5
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Entrez GeneBUB1B (a.k.a. BUB1beta, BUBR1, Bub1A, MAD3L, MVA1, SSK1, hBUBR1)
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Tags
/ Fusion Proteins
- GFP (N terminal on insert)
- AID (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site Not1 (not destroyed)
- 5′ sequencing primer ACT AGT GGA TCC CTA TCA CTG AAA GAG CAA AGC CCC AGG ACT AGT TAA CT
- 3′ sequencing primer ACT CGA GCG GCC GCC TAT CAC TGA AAG AGC AAA GCC CCA GGA CTA GTT AA
- (Common Sequencing Primers)
Resource Information
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Addgene Notes
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A portion of this plasmid was derived from a plasmid made byThe AID ORF was a kind gift from Masato Kanemaki (National Institute of Genetics, Mishima, Japan).
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
Nishimura et al., 2009, Nature Methods
DNA (Catalog # 47330-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 # 47330-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:
pcDNA5-EGFP-AID-BubR1 was a gift from Don Cleveland (Addgene plasmid # 47330 ; http://n2t.net/addgene:47330 ; RRID:Addgene_47330) -
For your References section:
Inducible, reversible system for the rapid and complete degradation of proteins in mammalian cells. Holland AJ, Fachinetti D, Han JS, Cleveland DW. Proc Natl Acad Sci U S A. 2012 Dec 4;109(49):E3350-7. doi: 10.1073/pnas.1216880109. Epub 2012 Nov 12. 10.1073/pnas.1216880109 PubMed 23150568