OGS394

PSF-CMV-PGK-PURO - DUAL PROMOTER PUROMYCIN PLASMID

plasmid vector for molecular cloning

• 別名: クローニングベクター, プラスミド, プラスミドベクター, ベクター, 分子クローニングベクター, 発現ベクター
• UNSPSCコード: 12352200
• NACRES: NA.85

特性

• 形状: buffered aqueous solution
• 分子量: size 5346 bp
• 微生物選択: kanamycin
• 哺乳類細胞選択: puromycin
• 複製起点: pUC (500 copies)
• ペプチド切断: no cleavage
• プロモーター: Promoter name: CMV; Promoter activity: constitutive; Promoter type: mammalian
• レポーター遺伝子: none
• 輸送温度: ambient
• 保管温度: −20°C

詳細

詳細

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.

アプリケーション

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.

シーケンス

To view sequence information for this product, please visit the product page

アナリシスノート

To view the Certificate of Analysis for this product, please visit www.oxgene.com

安全性情報

• 保管分類コード: 12 - Non Combustible Liquids
• 引火点(°F): Not applicable
• 引火点(℃): Not applicable