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Alternate Multiple Cloning Site

OG106 - pSF-CMV-pUC19

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CMV Expression Plasmid with the pUC19 MCS

pSF-CMV-pUC19

This plasmid contains the CMV promoter upstream of the multiple cloning site from pUC19.           

 

 

Description

Full Plasmid Details (PDF)

Sequence (txt)

This plasmid is compatible with all of the other DNA sequences available on this site, which are all sold in the same backbone. This flexibility allows you to create any vector you require, with a variety of functions, simply by compiling the relevant sections from our product range.

Product Code: OG106

Size (bp) 4309

Bacterial Antibiotic Selection: Ampicillin

Origin and Compatibility: pUC high copy, derived from pBR322

Copy Number: 500-700 copies per cell

Promoter: Cytomegalovirus (CMV) immediate early promoter

Purpose: A versatile cloning plasmid for the expression of genes in mammalian cells. This plasmid contains the multiple cloning site (MCS) from pUC19, however, it has been modified slightly to accommodate some restriction sites in the SnapFastTM system. These changes are described in the full plasmid details PDF. The use of this MCS, instead of the normal SnapFastTM vector MCS, will limit the ability to use some of the inserts that we sell that immediately flank, or are inserted within, the standard MCS. This primarily includes N-terminal tags and signal peptides. This is because these inserts are flanked by restriction sites that are not compatible with the pUC19 MCS. Most other inserts should still be compatible with this plasmid.

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.

Inserting a Gene: Each restriction site in this vector has a purpose, and allows the insertion of specific pre-designed DNA sequences. However, the presence of the pUC19 MCS in this plasmid, rather that our standard MCS, will make some of our products incompatible. If you would like to learn more about the restriction sites that are shared between this plasmid and our other plasmids please click here.

Intellectual Property Status: According to our IP-friendly policy, this plasmid is sold free of reach-through rights on any derivatives you may create.

Quality Sequencing Analysis: This plasmid has been fully sequenced using primers F1-F10. These are available through our website using the sequencing primers link on the left hand lower menu.

Genetic Modifications to standard parts:

CMV promoter – an NcoI site has been ablated in the CMV promoter. This change does not reduce expression.

AmpR cassette – This AmpR resistance region is not a wild type sequence, it has been modified to remove all restriction sites that conflict with the SnapFast system and to retain a high level of antibiotic resistance. It has been validated in E.coli.

Restriction site notes: 

  • Bgl2, KpnI, SwaI and PmeI each cut the plasmid at two sites positioned to flank the promoter, origin, and AmpR, respectively.
  • BsgI and BseRI recognise non-palindromic DNA sequences and cleave upstream of their recognition sites, within the stop codon that is found inside the XbaI restriction site.