pGLuc Mini-TK 2 Vector

This product has been discontinued and replaced by N8082.

  • This product was discontinued on 01/01/2017

Ordering Information

N8086
  • Discontinued
    Discontinued
  • Product Information
    pGLuc Mini-TK 2 is a cloning vector for mammalian cells, containing a minimal promoter fragment from the HSV thymidine kinase (TK) promoter adjacent to a reporter gene, the secreted luciferase from the copepod Gaussia princepsGaussia Luciferase (GLuc) is a 19 kDa protein encoded by a "humanized" sequence, and it contains a native signal peptide at the N-terminus that allows it to be secreted from mammalian cells into the cell culture medium (1,2). The pGLuc Mini-TK 2 Vector contains a multiple cloning site (MCS) upstream of the minimal TK promoter for cloning promoter or enhancer elements. A neomycin resistance gene under the control of the SV40 promoter allows selection for stable integration of the plasmid into the mammalian cell genome using G418.

    Figure 1: Figure 1
     A genomic DNA fragment containing putative estrogen response elements was inserted into the polylinker of pGLuc Mini-TK 2 Vector. The properties of this fragment were assessed in a transfection assay. GLuc activity was measured in the culture supernatant of MCF-7 cells transfected with the construct
    and treated with vehicle control (C) or estradiol (E2).

    Figure 2: The pGLuc Mini-TK 2 Vector MCS
    Figure 2: The pGLuc Mini-TK 2 Vector MCS
    Figure 2: The pGLuc Mini-TK 2 Vector MCS
    The TATA box is underlined and the first amino acids of GLuc are indicated above the DNA sequence.

    DNASU and Addgene are central repositories for plasmid clones and collections that may also be helpful.

    Highlights

    • Polylinker MCS: 12–68
    • Minimal promoter from Herpes simplex virus Thymidine Kinase (Mini-TK): 69–131
    • GLuc coding: 146–703
    • Start codon: 146–148
    • Stop codon: 701–703 • Signal peptide: 146–196 • Synthetic poly-A site: 712–760
    • Neo promoter (SV40): 1346-1681
    • Neomycin resistance gene: 1733-2527
    • Bacterial replication ori (pMB1): 3861-3273
    • Amp resistance: 4892-4032
    • All pGLuc-2 vectors have improved polyadenylation- transcription termination of the luciferase transcript. The polyadenylation signal is a synthetic polyadenylation sequence based on the b-globin gene (5).

    Product Source

    Isolated from E. coli strain NEB 10-beta by a standard DNA purification procedure.
    Product Categories:
    Gaussia Luciferase Products,
    DNA Plasmid Products
    Applications:
    Gaussia Luciferase
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