• All pGLuc-2 vectors have improvedpolyadenylation-transcription termination of theluciferase transcript. The polyadenylation signal isa synthetic polyadenylation sequence based on theβ-globin gene (5).
• Start codon: 920–922
• Stop codon: 1475–1477
• Signal peptide: 920–970
• Synthetic poly-A site: 1486–1534
• Neo promoter (SV40): 2120–2455
• Neomycin resistance gene: 2507–3301
• Bacterial replication ori (pMB1): 4635–4047
• Amp resistance: 5666–4806
Description:
The pCMV-GLuc 2 Control Plasmid is a mammalian expression vector that encodes the secreted luciferase from the copepod Gaussia princeps as a reporter, under the control of the constitutive CMV (cytomegalovirus) promoter. Gaussia luciferase (GLuc) is a 12 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). A neomycin resistance gene under the control of an SV40 promoter allows selection for stable integration of the plasmid into the mammalian cell genome using G418.
Recommended Sequencing Primers for pCMV-GLuc 2Control Plasmid (not available from NEB)
T7 Universal Primer (20-mer) TAATACGACTCACTATAGGG (863–882)
pBasic Reverse Primer (25-mer) TCAGAAGCCATAGAGCCCACCGCAT (1629–1605)
GLuc 3´ End Forward Primer (20-mer) GCCAGCAAGATCCAGGGCCA (1424–1443)
GLuc 5´ End Reverse Primer (24-mer) TCAGGGCAAACAGAACTTTGACTC (947–924)
Source:
Isolated from E. coli strain NEB10β by a standard DNA purification procedure.
Concentration:
500 μg/ml
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C
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