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.
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.