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Depositing Lab
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Depositing OrganizationUniversity of Cambridge
Located in United Kingdom of Great Britain and Northern Ireland -
Publication
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Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 26061 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 26061-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 26061-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepCMX
- Backbone size w/o insert (bp) 4500
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Vector typeMammalian Expression
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 nameCyclin B1
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SpeciesH. sapiens (human)
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Entrez GeneCCNB1 (a.k.a. CCNB)
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Tag
/ Fusion Protein
- GFP (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (unknown if destroyed)
- 3′ cloning site BamHI (unknown if destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer GFP-R
- (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
The stop codon of cyclin B1 in the pCMX vector was mutated to a Mfe1 site to link cyclin B1 to MmGFP. Cyclin B1 was fused to the second amino acid residue of MmGFP by a 5 amino acid linker (AGAQF). The construct is cloned as a HindIII-BamHI fragment into pCMX.
The Cyclin insert contains a V5A polymorphism which has no functional consequence. More information on MmGFP can be found in the following reference: Zernicka-Goetz, M., Pines, J., M., Ryan, K., Siemering, K.R., Haseloff, J., and Gurdon, J.B. (1996) ‘An indelible lineage marker for Xenopus using a mutated green fluorescent protein.
DNA (Catalog # 26061-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 # 26061-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:
pCMX/cyclinB1-GFP (27) was a gift from Jonathon Pines (Addgene plasmid # 26061 ; http://n2t.net/addgene:26061 ; RRID:Addgene_26061) -
For your References section:
Translocation of cyclin B1 to the nucleus at prophase requires a phosphorylation-dependent nuclear import signal. Hagting A, Jackman M, Simpson K, Pines J. Curr Biol. 1999 Jul 1. 9(13):680-9. 10.1016/S0960-9822(99)80308-X PubMed 10395539