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Multiple Cloning Site to Insert a Promoter

OG402 - No-Promoter Multiple Cloning Site Plasmid -pSF-PromMCS

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No-Promoter Multiple Cloning Site Plasmid -pSF-PromMCS

This plasmid is ideal for inserting new promoters upstream of the main MCS.       

 

Featuring SnapFastTM Technology 

 

 

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: OG402

Size (bp) 3806

Bacterial Antibiotic Selection: Kanamycin

Origin and Compatibility: pUC high copy, derived from pBR322

Copy Number: 500-700 copies per cell

Promoter: Not applicable

Purpose: This vector does not contain a promoter upstream of the main multiple cloning site. Instead this vector contains a short multiple cloning site (SalI to BstBI) in the same position that all of our promoters are normally inserted (flanked by Bgl2 sites). This plasmid is therefore ideal for inserting new promoters.

Poly A signals: Unlike pSF-pA-PromMCS, this plasmid does not contain a synthetic polyadenylation signal immediately upstream of the promoter multiple cloning site to reduce background transcription into the promoter MCS. This makes this plasmid useful when transferring any promoter into retroviral vector because the presence of a polyA can prevent full length virus genome production by causing premature transcription termination.  

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 grows to high titer in Escherichia coli using Ampicillin at a final concentration of 50µg/ml.

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:

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 region, 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.