LEGO-GM55
(Plasmid #217413)

• Purpose: Luciferase expression vector with the minimal GM-CSF promoter
• Depositing Lab: Peter Cockerill
• Publication: Cockerill Lab Luciferase Plasmids 2024 (unpublished)

Backbone

• Vector backbone: LeGO-iG
• Backbone manufacturer: Boris Fehse (Addgene #27358)
• Vector type: Mammalian Expression, Lentiviral, Luciferase

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Luciferase, GFP
• Alt name: SV40 PolyA sites, Human -55 to +28 minimal CSF2 promoter, Promega pGL3 LUC+, IRES-EGFP
• Species: H. sapiens (human); Firefly
• Promoter: Human -55 to +28 minimal CSF2 promoter

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: n/a

Resource Information

• Supplemental Documents:
  • LEGO-GM55.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This is a minimal activity lentiviral luciferase expression vector that contains the human -55 to +28 CSF2 promoter which can be used as a vehicle for testing the activities of other inducible or tissue-specific DNA elements. This vector also has 2 copies of the SV40 transcription termination/polyadenylation signal upstream of the multiple cloning site, which blocks background read-through transcription originating in the vector backbone, which itself can be substantial (Bert et al, 2000). LEGO-GM55 contains the vector backbone from LeGO-iG (Weber et al, 2008) (Addgene #27358) and was made by deleting an array of 3 AP-1 sites from the plasmid LEGO-AP1x3-GM55. The inserted luciferase gene and SV40 polyA site sequences originate from the plasmid pXPG (Bert et al, 2000) (Addgene #71248). The Luc+ firefly luciferase gene originates from the plasmid pGL3 (Promega).
References:
Bert et al. Plasmid. 2000 Sep;44(2):173-82. PMID 10964627.
Weber et al. Mol Ther. 2008 Apr. 16(4):698-706. PMID 18362927.