pYUBcLIC-GFP
(Plasmid #183509)

• 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 mycobacteria.
• Depositing Lab: Martin Högbom
• Publication: Grave et al Protein Expr Purif. 2022 Oct;198:106132. doi: 10.1016/j.pep.2022.106132. Epub 2022 Jun 21.

Backbone

• Vector backbone: pYUBcLIC-GFP
• Backbone manufacturer: Matthew Bennett
• Backbone size (bp): 5611
• Vector type: Bacterial Expression
• Promoter: T7 lac
• Tags / Fusion Proteins:
  • folding reporter GFP (C terminal on insert)
  • 10x His-tag (C terminal on insert)

Growth in Bacteria

• Bacterial Resistance(s): Hygromycin, 200 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

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

Resource Information

• Supplemental Documents:
  • pYUBcLIC-GFP_GeneBank.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

pYUB vector backbone originates from pYUB28b (Addgene plasmid # 37277 ; http://n2t.net/addgene:37277 ; RRID:Addgene_37277).

Reference: Metabolic engineering of cofactor F420 production in Mycobacterium smegmatis. Bashiri G, Rehan AM, Greenwood DR, Dickson JM, Baker EN. PLoS One. 2010 Dec 29;5(12):e15803. doi: 10.1371/journal.pone.0015803 PubMed 21209917
****
LIC cassette together with folding reporter GFP and purification tag sequence 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