pCpGfree-vitro Hygromycin

Selectable CpG-free Plasmids - Hygromycin Resistance

pCpGfree-vitro Hygromycin is available with a CpG-free allele of the LacZ gene or a multiple cloning site (MCS). This MCS contains several commonly used restriction sites, for convenient cloning of a CpG-free gene or any open reading frame or cDNA.


CpG-free plasmid backbone

Selectable in E. coli and mammalian cells with hygromycin

pCpGfree-vitroHLacZ: a LacZ expression plasmid

pCpGfree-vitroHmcs: contains a multiple cloning site (MCS)

These products are covered by a Limited Use License (See Terms and Conditions).


• 20 μg of pCpGfree-vitroH plasmid provided as lyophilized DNA
E. coli GT115 strain provided lyophilized on a paper disk
• 4 pouches of E.coli Fast-Media® Hygro (2 TB and 2 Agar)


pCpGfree-vitro is a new family of expression vectors completely devoid of CpG dinucleotides that are selectable in mammalian cells. Similarly to the other pCpGfree plasmids (i.e. pCpGfree-lacZ, pCpGfree-mcs, and pCpGfree-siRNA), all the elements required for replication and selection of the plasmids in bacteria, and gene expression in mammalian cells have been modified to remove all CpG dinucleotides.

Composite CpG-free promoter combining the mouse CMV enhancer, the human elongation factor 1α core promoter and 5’UTR containing a synthetic intron (I 126). This composite promoter yields high and ubiquitous expression of the gene cloned into the mcs.
Multiple cloning site (MCS) or CpG-free allele of the lacZ gene. This reporter gene can be easily subcloned and replaced by a gene of interest.
CpG-free polyadenylation signals (pAn): The polyadenylation signals utilized are CpG-free versions of the SV40 late and human β-globin polyadenylation signals. These polyA enable efficient cleavage and polyadenylation reactions resulting in high levels of steady-state mRNA.
CpG-free matrix attached regions (MARs) are AT-rich sequences that are able to form barriers between independent expression cassettes.
• CpG-free Hygromycin resistance gene (hph-∆CpG): The CpG-free HygroR gene is active both in E. coli and mammalian cells.
CpG-free SV40 promoter works in tandem with a bacterial promoter located within a synthetic intron (I-EC2K). This composite promoter drives the expression of the resistance gene in both mammalian cells and E. coli.
CpG-free E. coli R6K gamma origin of replication: This origin is activated by the R6K specific initiator protein π, encoded by the pir gene. Expression of the pir gene is necessary for the replication and amplification of pCpGvitro plasmids. E. coli GT115 strain expresses a pir mutant gene that allows higher plasmid copy number.


pCpGfree-vitro backbone


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