pCpGfree

CpGfree is a family of plasmids compley devoid of CpG dinucleotides. Typically, the elements required for replication and selection of the plasmid in E. coli and gene expression in mammalian cells are rich in CpG. In the pCpGfree plasmids these elements are either naturally CpG-free, were modified to remove all CpGs, or entirely synthesized.

Specifications:
• pCpGfree-mcs contains a multiple cloning site (MCS)featuring several commonly used restriction sites, for convenient cloning of a CpG-free gene, such as the genes provided in the pSELECT plasmid or any open reading frame or  cDNA.
• pCpGfree-LacZ expresses the lacZΔCpG gene, a CpG-free allele of the LacZ gene.
• pCpGfree-mSEAP expresses the mSEAPΔCpG gene, a CpG-free allele of the mouse secreted embryonic alkaline phosphatase gene.
• pCpGfree-Lucia expresses a CpG-free allele of the novel synthetic secreted luciferase (Lucia) gene.

Selectable in E. coli with Zeocin™

These products are covered by a Limited Use License (See Terms and Conditions).
Contents:
20 μg of lyophilized DNA
- E. coli GT115 strain provided lyophilized on a paper disk
- 4 pouches of E.coli Fast-Media® Zeo (2 TB and 2 Agar)
Description:
CpG-free Plasmid Backbone
pCpGfree is a family of plasmids compley devoid of CpG dinucleotides.  Typically, the elements required for replication and selection of the plasmid in E. coli and gene expression in mammalian cells are rich in CpG. In the pCpGfree plasmids these elements are either naturally CpG-free, were modified to remove all CpGs, or entirely synthesized.

• Origin of replication: The E. coli R6K gamma ori has been modified to remove all CpGs. This origin is activated by the R6K specific initiator protein π, encoded by the pir gene [1].
• Bacterial promoter: EM2K is a CpG-free version of the bacterial EM7 promoter.
• Mammalian promoter: The CpG-free promoter combines the mouse CMV enhancer, the human elongation factor 1 alpha core promoter and 5’UTR containing a synthetic intron.
• Polyadenylation signal: The polyadenylation signal is a CpG-free form of the late SV40 polyadenylation signal.
• MAR: Matrix attached regions (MARs) are sequences typically AT-rich that are able to form barriers between independently regulated domains [2]. pCpG plasmids contain two MARs, from the 5’ region of the human IFN-β gene or β-globin gene that were chosen because they are naturally CpG-free. The MARs are placed between the bacterial and mammalian transcription units.
• Selectable marker: The Zeocin™ resistance gene is a small gene (<400 bp) that contains numerous CpG dinucleotides. A synthetic new allele was created that contains no CpGs.

Furthermore, all Dam methylation sites (GATC) have been removed to prevent prokaryotic methylation.

Provided with the E. coli GT115 Strain
Due to the presence of the R6K gamma origin of replication, pCpGfree plasmids can only be amplified in an E. coli mutant strain expressing a pir mutant gene. They will not replicate in standard E. coli strains. Therefore, pCpGfree plasmids are provided with the E. coli GT115 strain, a pir mutant also deficient in Dcm methylation.

Details:•2012 - Blood, 120(18):3699-3707
Cytosolic sensing of extracellular self-DNA transported into monocytes by the antimicrobial peptide LL37.
Chamilos G, Gregorio J, Meller S, Lande R, Kontoyiannis DP, Modlin RL, Gilliet M
 

pCpGfree-mcs

pCpGfree-LacZ

pCpGfree-Lucia

pCpGfree-mSEAP

InvivoGen pcpgf-promlc pCpGfree-promoter-Lucia 20 µg
InvivoGen kcpgf-sirna pCpGfree-siRNA 1 kit
InvivoGen kcpg-sirna2 pCpGfree-siRNADUO 1 kit
InvivoGen pcpgvtb-lz pCpGfree-vitroBLacZ 20 µg
InvivoGen pcpgvtb-mcsg2 pCpGfree-vitroBmcs 20 µg
InvivoGen pcpgvth-lz pCpGfree-vitroHLacZ 20 µg
InvivoGen pcpgvth-mcsg2 pCpGfree-vitroHmcs 20 µg
InvivoGen pcpgvtn-lz pCpGfree-vitroNLacZ 20 µg
InvivoGen pcpgvtn-mcsg2 pCpGfree-vitroNmcs 20 µg
InvivoGen pcpgr-mcs pCpGrich-mcs 20 µg
InvivoGen inh-pd32 PD0325901 2 mg
InvivoGen tlrl-pd98 PD98059 10 mg
InvivoGen pdn-hasc pDeNy-hASC 20 µg
InvivoGen pdn-hirak pDeNy-hIRAK 20 µg
InvivoGen pdn-hmyd88 pDeNy-hMyD88 20 µg
InvivoGen pdn-h pDeNy-h 20 µg
InvivoGen pdn-hnod2 pDeNy-hNOD2 20 µg
InvivoGen pdn-hpkr pDeNy-hPKR 20 µg
InvivoGen pdn-hrigi pDeNy-hRIG-I 20 µg
InvivoGen pdn-htirap pDeNy-hTIRAP 20 µg
InvivoGen pdn-htraf6 pDeNy-hTRAF6 20 µg
InvivoGen pdn-htram pDeNy-hTRAM 20 µg
InvivoGen pdn-htrif pDeNy-hTRIF 20 µg
InvivoGen pdn-masc pDeNy-mASC 20 µg
InvivoGen pdn-mcs pDeNy-mcs 20 µg
InvivoGen pdn-mmyd88 pDeNy-mMyD88 20 µg
InvivoGen pdv5-gfp-1 pDRIVE5-GFP-1 20 µg
InvivoGen pdv5-gfp-10 pDRIVE5-GFP-10 20 µg
InvivoGen pdv5-gfp-2 pDRIVE5-GFP-2 20 µg
InvivoGen pdv5-gfp-3 pDRIVE5-GFP-3 20 µg
InvivoGen pdv5-gfp-4 pDRIVE5-GFP-4 20 µg
InvivoGen pdv5-gfp-5 pDRIVE5-GFP-5 20 µg
InvivoGen pdv5-gfp-6 pDRIVE5-GFP-6 20 µg
InvivoGen pdv5-gfp-7 pDRIVE5-GFP-7 20 µg
InvivoGen pdv5-gfp-8 pDRIVE5-GFP-8 20 µg
InvivoGen pdv5-gfp-9 pDRIVE5-GFP-9 20 µg
InvivoGen pdrive5lc-afphafp pDRIVE5Lucia-AFP-hAFP 20 µg
InvivoGen pdrive5lc-ceahcea pDRIVE5Lucia-CEA/hCEA 20 µg
InvivoGen pdrive5lc-chef1 pDRIVE5Lucia-chEF1 20 µg
InvivoGen pdrive5lc-chef1ru5 pDRIVE5Lucia-chEF1/RU5' 20 µg
InvivoGen pdrive5lc-cmvferlef1 pDRIVE5Lucia-CMV-hFerL-chEF1 20 µg
InvivoGen pdrive5lc-hendoglin pDRIVE5Lucia-Endoglin 20 µg
InvivoGen pdrive5lc-hflt1 pDRIVE5Lucia-FLT1 20 µg
InvivoGen pdrive5lc-hafp pDRIVE5Lucia-hAFP 20 µg
InvivoGen pdrive5lc-hb29 pDRIVE5Lucia-hB29 20 µg
InvivoGen pdrive5lc-hbact pDRIVE5Lucia-hbAct 20 µg
InvivoGen pdrive5lc-hbactru5 pDRIVE5Lucia-hbAct/RU5' 20 µg
InvivoGen pdrive5lc-hcckar pDRIVE5Lucia-hCCKAR 20 µg
InvivoGen pdrive5lc-hcd14 pDRIVE5Lucia-hCD14 20 µg
InvivoGen pdrive5lc-hcd43 pDRIVE5Lucia-hCD43 20 µg
InvivoGen pdrive5lc-hcd45 pDRIVE5Lucia-hCD45 20 µg 

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