pCMV-Script BioVector NTCC质粒载体菌种细胞基因保藏中心

pCMV-Script BioVector NTCC质粒载体菌种细胞基因保藏中心
The pCMV-Script vector is derived from a high-copy-number pUC-based plasmid and is designed to allow protein expression in mammalian systems. Mammalian expression is driven by the human cytomegalovirus (CMV) immediate early promoter to promote constitutive expression of cloned inserts in a wide variety of cell lines. Selection is made possible in bacteria by the kanamycin-resistance gene under control of the prokaryotic β-lactamase promoter. The neomycin-resistance gene is driven by the SV40 early promoter, which provides stable selection with G418 in mammalian cells. 1 The pCMV-Script vector does not contain an ATG initiation codon. A translation initiation sequence must be incorporated if the DNA fragment to be cloned does not have an initiating ATG codon or an optimal sequence for initiating translation, such as the Kozak sequence [GCC(A/G)CCATGG].2
The multiple cloning site (MCS) contains fifteen unique restriction enzyme recognition sites organized with alternating 5´ and 3´ overhangs to allow serial exonuclease III/mung bean nuclease deletions. T3 and T7 RNA polymerase promoters flank the polylinker for in vitro RNA synthesis. The choice of promoter used to initiate transcription determines which strand of the DNA insert will be transcribed. The pCMV-Script vector can be rescued as single-stranded (ss) DNA. The plasmid contains a 454-nucleotide filamentous f1 phage intergenic region (M13-related) that includes the 307 bp origin of replication. The orientation of the f1 origin in pCMV-Script allows the rescue of antisense ssDNA by a helper phage. This ssDNA can be used for dideoxynucleotide sequencing (Sanger method) or site-directed mutagenesis.
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