pSF-CMV-Ub-Zeo AscI (OG23) Zeocin Selection Plasmid

Product Description

Zeocin resistance plasmid where the Zeocin resistance gene is driven by the Ubiquitin promoter away from the CMV expression cassette.

$220.92
Product code: OG23
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Plasmid Information

Product Name: pSF-CMV-Ub-Zeo_AscI

Product Code: OG23

Size (bp): 5927 bp

Bacterial Antibiotic Selection: KanR

Origin and Compatibility: pUC high copy derived from pBR322

Bacterial Copy Number: 500-700 per cell

Promoter: Cytomegalovirus (CMV) immediate early promoter / Human Ubiquitin Promoter

Plasmid Purpose:

This vector contains a Zeocin (Zeo) resistance expression cassette under control of the Ubiquitin (Ub) promoter that is designed as an add-on to any of our vectors. The entire Ub Zeo cassette is flanked by AscI site. This vector can be used to force recombination into the genome of mammalian cells following transfection and selection although this method can be inefficient. It can also be used conjunction with the SV40 origin of replication in cells expressing the SV40 large T antigen (such as 293T cells) to make stable cells lines following transfection and selection.

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. For this reason we stock a range of other promoters that are compatible with this plasmid and are available on request.

Vector Sequence Files

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.

Transcription Termination:

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 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.

Intellectual Property Status

According to our IP-friendly policy this plasmid is sold free of reach-through rights and can be used to make commercial products. However the plasmid itself (or derivatives) cannot be sold.