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Phage

OG37 - pSF-T7

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$307.00

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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: OG37

Size (bp): 3862

Bacterial Antibiotic Selection: Kanamycin

Origin and Compatibility: pUC high copy, derived from pBR322

Copy Number: 500-700 copies per cell

Promoter: T7 bacteriophage Promoter

Purpose: This plasmid allows transcription of un-polyadenylated and un-capped RNA using the T7 bacteriophage polymerase. Some in vitro transcription kits will enable the production of poly-adenylated and capped RNA using this vector, however, this will depend on the kit used. All SnapFastTM vectors contain a T7 terminator downstream of the multiple cloning site that will enable transcription termination. If required, the 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.

Transcription Termination: Although intended mainly for expression using T7 polymerase, 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 Validation: This plasmid has been demonstrated to express reporter genes to high levels following T7 in vitro transcription to produce capped mRNA, followed by RNA transfection into the lung carcinoma cell line, A549. The start codon of these reporters was positioned within the NcoI site and the stop codon was positioned within the XbaI site.

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 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.