pmKO-AF8
(Plasmid
#118224)
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PurposeKusabira- Orange-CENP-A expression plasmid
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Depositing Lab
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Depositing OrganizationOsaka Metropolitan University
Located in Japan -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 118224 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 118224-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 118224-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepmKO1-MC1
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Backbone manufacturerMBL, Nagoya, Aichi, Japan
- Backbone size w/o insert (bp) 4656
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Vector typeMammalian Expression
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameCENP-A
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Alt namecentromere protein A
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SpeciesH. sapiens (human)
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Entrez GeneCENPA (a.k.a. CENP-A, CenH3)
- Promoter CMV
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Tag
/ Fusion Protein
- Kusabira orange (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BglII/BamHI (destroyed during cloning)
- 3′ cloning site KpnI (not destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer SV40pA-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
Bgl II-KpnI fragment of pEGFP-AF8 (Addgene plasmid # 117803) was recloned into BamHI and KpnI site of pmKO1-MC1.
Note that the cloning strategy adds additional amino acids onto the C-terminus of the insert (STVPELQQRIRETREQSLNKRPRL), but this does not interfere with plasmid function. The extra sequence includes an MCS for adding tags to the C-terminus of the insert.
DNA (Catalog # 118224-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
-
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.
DNA (Catalog # 118224-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:
pmKO-AF8 was a gift from Kenji Sugimoto (Addgene plasmid # 118224 ; http://n2t.net/addgene:118224 ; RRID:Addgene_118224) -
For your References section:
Construction of three quadruple-fluorescent MDA435 cell lines that enable monitoring of the whole chromosome segregation process in the living state. Sugimoto K, Senda-Murata K, Oka S. Mutat Res. 2008 Nov 17;657(1):56-62. doi: 10.1016/j.mrgentox.2008.08.005. Epub 2008 Aug 19. 10.1016/j.mrgentox.2008.08.005 PubMed 18778791
