PBR325 Plasmid

The pBR325 plasmid is a classic cloning vector used in molecular biology. It is derived from the pBR322 plasmid, which itself is one of the first and most widely used plasmids for cloning purposes.

Benefits

• Multiple Antibiotic Resistance Genes: pBR325 contains three antibiotic resistance genes: the ampicillin resistance gene, the tetracycline resistance gene, and the chloramphenicol resistance gene. This feature allows for selective growth of bacteria containing the plasmid in the presence of these antibiotics and provides the flexibility to use different selection pressures depending on the experimental needs.
• Origin of Replication: pBR325 contains the ColE1 origin of replication, allowing it to replicate independently in a bacterial host, usually E. coli. The ColE1 origin ensures a moderate to high copy number of the plasmid within the bacterial cells.
• Multiple Cloning Site (MCS): pBR325 has several unique restriction enzyme sites for the insertion of foreign DNA. Cloning into some of these sites will disrupt the Tetracycline resistance gene, providing a means of identifying insertions through loss of tetracycline resistance.
• Size: The plasmid is 5996 bp, which facilitates its manipulation and transformation into bacterial cells.
• Compatibility with Common Laboratory Strains of E. coli: pBR325 can be easily introduced into and maintained in various lab strains of E. coli, making it a versatile tool for cloning and genetic engineering.

Product Specifications

Concentration: 0.2 μg/μL in TE
Volume: 50 μL
Backbone size: 5995 bp
Vector type: Cloning vector
Bacterial Resistance(s): Ampicillin, Tetracycline, Chloramphenicol
Copy number: High Copy
Genbank Assession Number: L08855

Vector Map

Sequence

GoldBio Vector Map and Sequence for pBR325

Note: There are several minor mutations in GoldBio pBR325 from what is reported in the Genbank sequence. These mutations include:

• Sequence position 1269: T for C (in Genbank) in the Tetracycline resistance gene. This changes the amino acid from Isoleucine to Threonine. This appears to have no effect on Tetracycline resistance.
• Sequence position 2027-2028: Missing T (Position 1893 in Genbank)
• Sequence position 2048-2049: Missing A (Position 1915 in Genbank)
• Sequence position 5397: Addition of T in a position downstream of the Chloramphenicol resistance gene. This is in a non-coding section of the vector and appears to have no effect on cloning.
• Sequence position 5831: A for G (in Genbank) in a position downstream of the Chloramphenicol resistance gene. This is in a non-coding section of the vector and appears to have no effect on cloning.

Storage/Handling

pBR325 DNA is at 0.2 μg/μL concentration in TE buffer.
Store all plasmids at -20°C.