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OG177 - pSF-T7-T7-Term


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Sequence (txt)

Vector: pSF-T7-T7 Term

Product Code: OG177

Quantity Provided: 5µg

Size (bp): 3448

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: This vector allows transcription of un-polyadenylated and un-capped RNA using the T7 bacteriophage polymerase. Some in vitro transcription kits will enable the production poly-adenylated and capped RNA using this vector, however, this will depend on the kit used. This vector contains the T7 terminator only, whereas most of our vectors contain mammalian, T7 and bacterial termination signals together. This plasmid is normally only used if the presence of the mammalian and bacterial terminators is undesirable.  

This vector can also be digested at a restriction site that is 3’ to the end of your gene to allow run-off of the polymerase, rather than exploiting the hairpin T7 terminator. If you are expressing your gene in a Vaccinia virus-free mammalian T7 system it will require an IRES upstream of your gene because of the absence of a 5’ cap on the RNA produced, please see OG85 – pSF-T7-EMCV if this is required.    

Transcription Termination: This plasmid contains only the T7 terminator after the multiple cloning site. The mammalian and bacterial terminators have been deleted. 

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. 

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:

  • 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 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 vector 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.