pETcLIC-GFP
(Plasmid
#183514)
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Purpose(Empty Backbone) The plasmid contains LIC cassette upstream of folding reporter GFP and TEV-cleavable 10x histidine purification tag. Compatible with T7 promoter-based expression system in E. coli
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 183514 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepETcLIC-GFP
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Backbone manufacturerMatthew Bennett
- Backbone size (bp) 5852
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Vector typeBacterial Expression
- Promoter T7 lac
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Tags
/ Fusion Proteins
- folding reporter GFP (C terminal on insert)
- 10x His-tag (C terminal on insert)
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 numberUnknown
Cloning Information
- Cloning method Ligation Independent Cloning
- 5′ sequencing primer T7 -for : TAA TAC GAC TCA CTA TAG GG
- 3′ sequencing primer T7-rev: CTA GTT ATT GCT CAG CGG T (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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
pET vector backbone originates from pETDuet-1 (Novagen).
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LIC cassette together with folding reporter GFP and purification tag sequences originate from pREcLIC-GFP.
Reference: Geertsma, E., Poolman, B. High-throughput cloning and expression in recalcitrant bacteria. Nat Methods 4, 705–707 (2007). doi: 10.1038/nmeth1073 PubMed 17643108
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:
pETcLIC-GFP was a gift from Martin Högbom (Addgene plasmid # 183514 ; http://n2t.net/addgene:183514 ; RRID:Addgene_183514) -
For your References section:
High-throughput strategy for identification of Mycobacterium tuberculosis membrane protein expression conditions using folding reporter GFP. Grave K, Bennett MD, Hogbom M. Protein Expr Purif. 2022 Oct;198:106132. doi: 10.1016/j.pep.2022.106132. Epub 2022 Jun 21. 10.1016/j.pep.2022.106132 PubMed 35750296