pEE14.1 GS expression vector高效表达载体GS加压筛选表达质粒 BioVector NTCC质粒载体菌种细胞基因保藏中心
- 价 格：¥498965
- 货 号：pEE14.1 GS expression vector高效表达载体GS加压筛选表达质粒
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pEE14.1 GS expression vector高效表达载体GS加压筛选表达质粒 BioVector NTCC质粒载体菌种细胞基因保藏中心
Map of pEE14 GS expression vector. pEE14 (~9.4 kb in length) contains a GS minigene as the selectable marker which has a single intron and GS polyadenylation signals and is driven from an SV40 late promoter. The hCMV-MIE promoter-enhancer and 5¢ untranslated region are used to express the gene of interest and the remainder of the plasmid contains an ampicillin-resistance gene and replication origin for replication in E. coli. The plasmid was constructed as follows. A 900-bp EcoRI fragment from the cDNA clone lgs1.1 (Hayward et al., 1986) was assembled with a 3.4-kb EcoRI-SacI hamster GS genomic fragment from pGS1 (Sanders and Wilson, 1984), which provides the 3¢ end of the minigene. (The SacI site was converted to a BamHI site to facilitate vector construction.) The EcoRI site within the GS coding sequence was destroyed by site-directed mutagenesis without altering the amino acid sequence and a HindIII site in GS 3¢-flanking DNA was destroyed by digestion with HindIII, filling in the single-stranded ends, and religation. A 340-bp SV40 late promoter (Cockett et al., 1990) was added to the 5¢ end as a BamHI-EcoRI fragment and the EcoRI site between the promoter and the GS sequences was destroyed by filling in. The resulting 4.5-kb BamHI fragment was inserted into pEE6hCMV (Stephens and Cockett, 1989) at a single BglII site upstream of the hCMV enhancer (hence destroying the BglII and BamHI sites) to form pEE14. The resulting SV40-GS minigene in pEE14 is functionally equivalent to that in pSVLGS.1 (Bebbington and Hentschel, 1987) but has been deleted of EcoRI and HindIII sites.
【Supplier来源】BioVector NTCC Inc.
TEL：+86-010-53513060
【Website网址】 http://www.biovector.net
Map of pEE14 GS expression vector. pEE14 (~9.4 kb in length) contains a GS minigene as the selectable marker which has a single intron and GS polyadenylation signals and is driven from an SV40 late promoter. The hCMV-MIE promoter-enhancer and 5¢ untranslated region are used to express the gene of interest and the remainder of the plasmid contains an ampicillin-resistance gene and replication origin for replication in E. coli. The plasmid was constructed as follows. A 900-bp EcoRI fragment from the cDNA clone lgs1.1 (Hayward et al., 1986) was assembled with a 3.4-kb EcoRI-SacI hamster GS genomic fragment from pGS1 (Sanders and Wilson, 1984), which provides the 3¢ end of the minigene. (The SacI site was converted to a BamHI site to facilitate vector construction.) The EcoRI site within the GS coding sequence was destroyed by site-directed mutagenesis without altering the amino acid sequence and a HindIII site in GS 3¢-flanking DNA was destroyed by digestion with HindIII, filling in the single-stranded ends, and religation. A 340-bp SV40 late promoter (Cockett et al., 1990) was added to the 5¢ end as a BamHI-EcoRI fragment and the EcoRI site between the promoter and the GS sequences was destroyed by filling in. The resulting 4.5-kb BamHI fragment was inserted into pEE6hCMV (Stephens and Cockett, 1989) at a single BglII site upstream of the hCMV enhancer (hence destroying the BglII and BamHI sites) to form pEE14. The resulting SV40-GS minigene in pEE14 is functionally equivalent to that in pSVLGS.1 (Bebbington and Hentschel, 1987) but has been deleted of EcoRI and HindIII sites.
【Supplier来源】BioVector NTCC Inc.
TEL：+86-010-53513060
【Website网址】 http://www.biovector.net
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