Plasmid Info:

Plasmid Purpose:

Frosty CFP (Cyan Fluorescence Protein) expression plasmid with a multiple cloning site upstream to allow your preferred promoter to be inserted. Also contains a poly A site upstream to reduce background readthrough in mammalian systems

Sequence and Map:

Other Info:

This plasmid contains three alternative transcription terminators for mammalian bacterial and bacteriophage (T7) expression. This means that only the promoter needs to be changed to alter the expression system you are using. We sell multiple promoters that can be used in each of these systems. The presence of each terminator does not reduce expression in the alternative systems.

Cloning:

This plasmid contains a gene within the main multiple cloning site (NotI-ClaI). Any plasmid that we sell where the gene is this configuration will be located in the exact same position in relation to the start and stop codon of the gene. The only exceptions to this rule are fusions proteins where the fusion gene may be positioned at the front or end of the MCS to allow gene fusion.

By positioning all of our genes in the same location it allows them to be transferred between plasmids using the same cloning method and restriction sites regardless of the plasmid being used from our product range. Inserting a new gene into this plasmid should be easily possible using a range of standard restriction enzyme sites that flank the gene currently in the vector.

In the multiple cloning site there are two important restriction sites called BsgI and BseRI sites. These sites both cut the DNA at the same position and cleave the stop codon of the gene in the multiple cloning site in this plasmid thereby producing a TA overhang. This overhang is compatible with any of our peptide or reporter fusion tag plasmids also cut with either of these enzymes. This allows seamless C-terminal fusions to be made with the gene in this multiple cloning site using a single cloning step from our C-terminal peptide and reporter tag product range. Normally the easiest method is to clone the C-terminal tag from our other plasmid products into this plasmid using BsgI or BseRI and the downstream ClaI restriction site.

BseRI and BsgI sites are non-palindromic and cleave a defined number of bases away from their binding sites. This allows them to cut the upstream stop codon in the gene in this plasmid regardless of the gene sequence.

IP Status:

This product is part of our SnapFast plasmid range, for more information on the Intellectual property status of this plasmid please click here

Oxford Genetics公司位于英国牛津，由来自生物各领域中具有多年基因工程经验的专家组建而成，专注于为全球生物不同领域的科研用户和生物技术工业用户提供多种DNA表达载体。 其所有产品基于一个中心质粒骨架(SnapFast )SnapFast 载体经过精心设计，不同的构建中配以不同的DNA元件，在特异酶切位点更换需要的元件，可进行上千种改造，满足各种生物学研究的需要。该公司还为科学家提 供万能克隆平台和DNA插入匹配库，有针对哺乳动物，细菌和噬菌体等多个系统的各种元件，包括启动子、报告基因、抗性标记、标签蛋白、核糖体位点、复制起 点、多克隆位点、融合蛋白、信号肽、终止子及其他DNA元件(SV40原点、P2A肽段、CMV增强子、小启动子等)。所有的元件都可以在不同载体间互换 使用，并且和很多市场上应用的克隆载体和穿梭载体都兼容，利于基因片段的转移。该系统非常灵活、高效、经济、知识产权友好，可解决目前基因工程研究中普遍 存在的克隆问题。