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Description_x000D_
General description_x000D_
This plasmid contains the major immediate early cytomegalovirus (CMV) enhancer region upstream of the human secreted alkaline phosphatase (SEAP) reporter gene. The enhancer can be used to increase the strength of weaker promoters whilst often retaining any regulation that they already have such as tissue specificity or transcription factor activation in response to stimulus. This enhancer sequence has been inserted upstream of a synthetic poly-adenylation signal that reduces non-specific transcription of any gene that is inserted into the multiple cloning site._x000D_
Promoter Expression Level:_x000D_
Application_x000D_
Cloning in a gene: 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._x000D_
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 sIt`s 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 sIt`s that flank the gene currently in the vector._x000D_
Multiple cloning site notes: In the multiple cloning site there are two important restriction sIt`s called BsgI and BseRI sIt`s. These sIt`s 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._x000D_
BseRI and BsgI sIt`s are non-palindromic and cleave a defined number of bases away from their binding sIt`s. This allows them to cut the upstream stop codon in the gene in this plasmid regardless of the gene sequence._x000D_
Sequence_x000D_
Quick-reference Plasmid Map_x000D_
Please select the file type you require. For reference most cloning programs will import a .gb (Genbank) file and will show all of the plasmids features automatically when downloaded and imported._x000D_
Genebank Vector Sequence File_x000D_
FASTA Vector Sequence File_x000D_
Full Plasmid Map_x000D_
Analysis Note_x000D_
To view the Certificate of Analysis for this product, please visit www.oxfordgenetics.com._x000D_
Other Notes_x000D_
Looking for more vector options to move your experiments forward faster/ Consider a custom cloning vector designed and built by Oxford Genetics™. Find out more at Oxford Genetics - Sigma's partner for cloning and expression vectors for molecular biology and synthetic biology applications._x000D_
Legal Information_x000D_
Oxford Genetics is a trademark of Oxford Genetics Ltd
- Related Categories Cloning and Expression, Molecular Biology, SnapFast Cloning Vectors, SnapFast Vectors for Mammalian Host (untagged) form