pET22b-HA-CPDSalI
(Plasmid
#38253)
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Purpose(Empty Backbone)
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Depositing Labs
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 38253 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET22b
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Backbone manufacturerNovagen
- Backbone size (bp) 6153
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Modifications to backboneHA epitope tag and Vibrio cholerae MARTX toxin (rtxA, vc1451) cysteine protease domain (CPD) (amino acids 3440-3650) cloned into SalI and XhoI sites in backbone to create fusions with target protein of interest
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Vector typeBacterial Expression
- Promoter T7
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Tags
/ Fusion Proteins
- HA (C terminal on backbone)
- Vibrio cholerae MARTX toxin cysteine protease domain (CPD) (C terminal on backbone)
- 6xHis (C terminal on backbone)
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 numberLow Copy
Cloning Information
- Cloning method Restriction Enzyme
- 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.
Depositor Comments
For protein purification, target protein of interest should be cloned into NdeI and SalI sites to create HA-tagged fusion with Vibrio cholerae MARTX toxin cysteine protease domain (CPD). HA epitope tag will remain on target protein after cleavage.
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:
pET22b-HA-CPDSalI was a gift from Matthew Bogyo & Aimee Shen (Addgene plasmid # 38253 ; http://n2t.net/addgene:38253 ; RRID:Addgene_38253) -
For your References section:
Simplified, enhanced protein purification using an inducible, autoprocessing enzyme tag. Shen A, Lupardus PJ, Morell M, Ponder EL, Sadaghiani AM, Garcia KC, Bogyo M. PLoS One. 2009 Dec 2;4(12):e8119. 10.1371/journal.pone.0008119 PubMed 19956581