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PurposeExpresses GFP with +9 theoretical charge in E.Coli
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Depositing Lab
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Sequence Information
Ordering
This material is available to academics and nonprofits only.
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 89247 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
Backbone
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Vector backbonepET29
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Backbone manufacturerNovagen
- Backbone size w/o insert (bp) 5152
- Total vector size (bp) 5896
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Vector typeBacterial 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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Growth instructionsCan also maintain in Mach1. For protein expression, transform into BL21 Star (DE3) cells. Induce at 0.6 OD with 0.5 mM IPTG final concentration. Growing overnight (16 hours) at 20-25 C.
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameHis6-(pos9)-GFP
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SpeciesSynthetic
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Insert Size (bp)745
- Promoter T7
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Tag
/ Fusion Protein
- His6 (N terminal on insert)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer T7 Promoter
- 3′ sequencing primer T7 Terminator (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.
How to cite this plasmid
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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:
pET-6xHis-(pos9)GFP was a gift from David Liu (Addgene plasmid # 89247 ; http://n2t.net/addgene:89247 ; RRID:Addgene_89247) -
For your References section:
Cellular uptake mechanisms and endosomal trafficking of supercharged proteins. Thompson DB, Villasenor R, Dorr BM, Zerial M, Liu DR. Chem Biol. 2012 Jul 27;19(7):831-43. doi: 10.1016/j.chembiol.2012.06.014. 10.1016/j.chembiol.2012.06.014 PubMed 22840771
