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Purpose(Empty Backbone) SGC Empty backbone for bacterial expression
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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 | 26107 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepET-28-a
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Backbone manufacturerNovagen
- Backbone size (bp) 5515
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Vector typeBacterial 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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Growth instructionsFor plasmid propagation and cloning, use any E. coli strain that does not express T7 RNA polymerase. For expression, use host cells that express T7 RNA pol, e.g. BL21(DE3).
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameNone
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GenBank IDGU452710
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Tag
/ Fusion Protein
- His6 - Z-basic - TEV (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site Ligation-Independent cloning; cut with BsaI (destroyed during cloning)
- 3′ cloning site Ligation-Independent cloning; cut with BsaI (destroyed during cloning)
- 5′ sequencing primer T7F
- 3′ sequencing primer T7R (Common Sequencing Primers)
Resource Information
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Article 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.
Depositor Comments
Primers for LIC cloning:
Upstream: add TACTTCCAATCCATG to the 5’ end (ATG in-frame with the desired coding
sequence).
Downstream: add TATCCACCTTTACTG to 5’ end of downstream primer; add termination
codon, if necessary.
Detailed cloning method available in the paper (Savitsky et al., J Struct Biol., 2010)
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:
pNIC-ZB was a gift from Opher Gileadi (Addgene plasmid # 26107 ; http://n2t.net/addgene:26107 ; RRID:Addgene_26107) -
For your References section:
High-throughput production of human proteins for crystallization: The SGC experience. Savitsky P, Bray J, Cooper CD, Marsden BD, Mahajan P, Burgess-Brown NA, Gileadi O. J Struct Biol. 2010 Jun 10. ():. 10.1016/j.jsb.2010.06.008 PubMed 20541610