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SV40 Origins

OG114 - pSF-CMV-SV40 Ori Sbfl

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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: OG114

Size (bp): 4436

Bacterial Antibiotic Selection: Kanamycin

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 vector for the expression of genes in mammalian cells. This vector also contains the origin of replication from Simian Virus 40 (SV40), which enables it to be replicated in cells that express the SV40 large T antigen (such as 293T cells). The SV40 origin can be used to increase the level of transcription in cells that are transfected with the plasmid because the plasmid is replicated. It is also reported that by using selectable markers in conjunction with the SV40 origin, stable cell lines can be produced. However, we have not validated it for this purpose. We currently sell a range of antibiotic selection genes that can be used for this purpose, in a range of positions within the SnapFastTM system. In this vector, the SV40 origin is inserted within the SbfI site, and is flanked by two SbfI sites.      

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. To insert a gene, we recommend using any sites between (and including) the HindIII and XbaI sites. By doing this you will be able to insert most of our other products upstream, or downstream, of your gene. Click here to read more about the purpose of each restriction site.

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 vector has been fully sequenced using primers F1-F10. The sequences and details of these primers are available through our website. 

Genetic Modifications to standard parts:

  • CMV promoter – an NcoI site has been ablated in the CMV promoter. This change does not reduce expression.
  • KanR cassette – This KanR resistance region is not a wild type sequence, it has been modified to remove all restriction sites that conflict with the SnapFastTM 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, start codon, origin, and KanR, 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.
  • The SV40 origin in this vector contains multiple restriction sites, including NcoI, BamHI, ClaI and BseRI sites, as well as two StuI sites. These sites are currently being ablated, when completed; the ablated version will replace this vector. If you would prefer the vector without these sites, please contact us, as it may be nearing completion.