形狀
buffered aqueous solution
分子量
size 5346 bp
菌種選擇
kanamycin
哺乳動物細胞選擇
puromycin
複製起點
pUC (500 copies)
肽切割
no cleavage
啟動子
Promoter name: CMV
Promoter activity: constitutive
Promoter type: mammalian
報告基因
none
運輸包裝
ambient
儲存溫度
−20°C
一般說明
This plasmid contains two promoters that terminate transcription at the same poly-adenylation signal allowing the expression of two genes from one expression cassette where the second gene is the puromycin resistance gene (puro) for antibiotic selection in mammalian cells. The second promoter (PGK) is approximately 10-fold weaker than the upstream CMV promoter in most commonly used cell lines.
Promoter Expression Level: This plasmid contains the mammalian CMV promoter to drive gene expression. We have tested all of our mammalian promoters in a range of cell types and CMV is consistently the strongest in those we have studied. However there are many reports of the CMV promoter demonstrating silencing by methylation in long-term culture.
Promoter Expression Level: This plasmid contains the mammalian CMV promoter to drive gene expression. We have tested all of our mammalian promoters in a range of cell types and CMV is consistently the strongest in those we have studied. However there are many reports of the CMV promoter demonstrating silencing by methylation in long-term culture.
應用
Cloning in a gene: This plasmid has been designed to be compatible with a range of cloning techniques. The multiple cloning site contains a range of standard commonly used restriction sites for cloning. Using these sites genes can be inserted using standard cloning methods with DNA ligase. Other methods such as ligase independent cloning (LIC) Gibson Assembly InFusionHD or Seamless GeneArt can also be used and because all of our plasmids are based on the same backbone the same method can be used for cloning into all of our catalogue vectors.
Multiple cloning site notes: There are a few important sites within the MCS. These include the NcoI site the XbaI site and the BsgI and BseRI sites. The NcoI site contains a start codon that is immediately downstream of both a Kozak and Shine-Dalgarno ribosomal binding site. These allow for optimal positioning of genes when the start codon is placed in this location. If this is not required and you wish to use a downstream site for gene cloning you can remove the NcoI site by cleaving the plasmid with KpnI.
The XbaI site contains a stop codon. This stop codon is positioned in a specific position in relation to the BsgI and BseRI sites that are immediately downstream. When either BseRI or BsgI cleave the plasmid they produce a TA overhang from the stop codon in the XbaI site that is compatible with all of our peptide tag plasmids cut with the same sites. BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding site.
Whenever we clone a gene into our multiple cloning site we always position the start and stop codon in the same positions in the MCS. If the start and ends of the genes are not compatible with NcoI and XbaI we extend the sequence to the nearest external sites but keep the start and stop codons locations consistent.
Multiple cloning site notes: There are a few important sites within the MCS. These include the NcoI site the XbaI site and the BsgI and BseRI sites. The NcoI site contains a start codon that is immediately downstream of both a Kozak and Shine-Dalgarno ribosomal binding site. These allow for optimal positioning of genes when the start codon is placed in this location. If this is not required and you wish to use a downstream site for gene cloning you can remove the NcoI site by cleaving the plasmid with KpnI.
The XbaI site contains a stop codon. This stop codon is positioned in a specific position in relation to the BsgI and BseRI sites that are immediately downstream. When either BseRI or BsgI cleave the plasmid they produce a TA overhang from the stop codon in the XbaI site that is compatible with all of our peptide tag plasmids cut with the same sites. BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding site.
Whenever we clone a gene into our multiple cloning site we always position the start and stop codon in the same positions in the MCS. If the start and ends of the genes are not compatible with NcoI and XbaI we extend the sequence to the nearest external sites but keep the start and stop codons locations consistent.
序列
To view sequence information for this product, please visit the product page
分析報告
To view the Certificate of Analysis for this product, please visit www.oxgene.com
儲存類別代碼
12 - Non Combustible Liquids
閃點（°F）
Not applicable
閃點（°C）
Not applicable
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商品
Learn more about relevant restriction site functions in the SnapFast™ plasmid system. All DNA sections are pre-screened, and where possible modified, to remove any of the restriction sites found within the core SnapFast plasmids to maintain their flexibility.
A range of forward and reverse sequencing primers that allow you to sequence any insert that you make into a particular position within any plasmid. Where possible, the binding sites for each of these primers is conserved.
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