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pMOD2 bearing bacterial synthetic Pac gene
Resistance to the antibiotic puromycin is conferred by the Pac gene encoding a puromycin N-acetyl-transferase isolated from a Streptomyces strain. The DNA sequence of this gene was modified by optimizing the codon usage and reducing CpG motifs to 16 without changing the amino acid sequence of the wild type protein.
The low-CpG Pac gene is provided in the bacterial plasmid pMOD2 which is selectable with Ampicillin. To facilitate the excision and subcloning of the low-CpG Pac gene into another vector, this gene is flanked by a unique restriction site at the 5’ and 3’ end.
Gene: Low-CpG Pac
Description: Synthetic Puromycin resistance gene
93 CpG in the native gene, 16 CpG in the synthetic geneBack to the top
• 20 µg of lyophilized pMOD2-Puro plasmid DNA
• 4 pouches of E. coli Fast-Media® Amp (2 LB and 2 Agar)
• Multiple cloning sites
MCS1, located upstream of the synthetic gene, contains the following restriction sites:
Nde I, Bst EII, Avr II, Mfe I, Bgl II, Hind III, Pme I, Bsp HI
MCS2, located downstream of the synthetic gene, contains the following restriction sites:
Eco RI, Bam HI, Nhe I, Eco RV, Sac I, Pac I
• Synthetic Puromycin resistance gene [Puro-lowCpG]: The Pac gene encoding a puromycin N-acetyl-transferase has been isolated from a Streptomyces strain. Its expression confers puromycin resistance to pac-transfected cells. The size of the Pac gene is small but its CpG content is very high (93 CpGs for a total of 597 bp). The number of CpG dinucleotides has been reduced to 16 and the codon usage optimized.
• rnpb term: The E. coli rnpb terminator allows efficient transcription termination of the Puro-lowCpG gene.
• pMB1 Ori: To limit vector size, InvivoGen uses a minimal E. coli origin of replication with the same activity as the longer Ori.
• Amp: The ampicillin resistance gene allows the selection of transformed E. coli carrying a pMOD2 plasmid.