Gateway® entry vectors are designed to clone DNA sequences using restriction endonucleases and ligase to create a Gateway® entry clone. The resulting entry clone is ready for recombination with a destination vector to create an expression clone.
The Gateway® entry vectors (Table 1) offer the following:
• attL1 and attL2 sites for site-specific recombination of the entry clone with a Gateway® destination vector to ensure cloning of the gene of interest in the correct orientation for expression
• Kozak consensus sequence for efficient translation initiation in eukaryotic systems
• Ribosome binding site for efficient translation initiation in prokaryotic systems (pENTR™ 1A Dual Selection, pENTR™3C Dual Selection, and pENTR™11 Dual Selection vectors only)
• rrnB transcription termination sequences to prevent basal expression of the PCR product of interest in E. coli
• pUC origin for high-copy replication and maintenance of the plasmid in E. coli
• Kanamycin resistance gene for selection in E. 查看更多coli
• The ccdB⁄chloramphenicol fusion gene located between the two attL sites for
o negative selection and
o Chloramphenicol selection in E. coli
• Kanamycin resistance gene for selection in E. coli