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Purpose(Empty Backbone)
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Depositing Lab
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Publication
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 39264 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET26b(+)
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Backbone manufacturerEMD Biosciences
- Backbone size (bp) 5438
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Modifications to backbonepSANG10 was created by digesting pET26b(+) at the single XhoI site and at the BglI sites closest to the T7 promoter (by partial digestion) and replacing the fragment with a double stranded oligonucleotides compatible with these sites, using four annealed oligonucleotides (see Table 2 of manuscript for all oligonucleotide sequences). This introduced NcoI, PstI, NotI, HindIII restriction sites and a serine (pSANG10) immediately after the NotI site. The tri-FLAG epitope tag was inserted into pSANG10 at the HindIII site using two annealed oligonucleotides to create pSANG10-3F. The scFv encoding gene is sub-cloned at the NcoI/NotI site and the 5' HindIII site is available for insertion of peptide tags and fusion partners.
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Vector typeBacterial Expression ; Bacterial expression, bacterial expression of recombinant antibody fragments in the form of single chain Fvs (scFvs)
- Promoter T7
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Tags
/ Fusion Proteins
- 6xHis (C terminal on backbone)
- tri-FLAG (C terminal on backbone)
- pelB leader (N terminal on backbone)
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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Growth instructionsMust be transformed into BL21(DE3) bacteria and grown at 30 degrees for antibody production.
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Copy numberUnknown
Cloning Information
- Cloning method Restriction Enzyme
- 5′ sequencing primer T7 promoter
- 3′ sequencing primer T7 terminal primer (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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Articles Citing this Plasmid
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.
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:
pSANG10-3F was a gift from John McCafferty (Addgene plasmid # 39264 ; http://n2t.net/addgene:39264 ; RRID:Addgene_39264) -
For your References section:
A simple vector system to improve performance and utilisation of recombinant antibodies. Martin CD, Rojas G, Mitchell JN, Vincent KJ, Wu J, McCafferty J, Schofield DJ. BMC Biotechnol. 2006 Dec 7;6:46. 10.1186/1472-6750-6-46 PubMed 17156422