The Thermo Scientific pUC18 vector is a smaller, high-copy-number E. coli plasmid in length 2686 bp. It contains the same multiple cloning site (MCS) as the pUC19 vector, but is arranged in the opposite direction.

Product Advantages

• Purified by chromatography using proprietary patented technology
• More than 90% supercoiled form
• From Escherichia coli (dam+, dcm+)
• For pUC18 DNA sequence, pUC19 DNA sequence, sequence analysis and map creation, please See Free online REviewer tool.

Applications

• Cloning
• Insert DNA sequencing, pUC18 DNA: Preparation of DNA molecular weight standards

pUC18/19 Plasmid Contents and Usage Notes - The pUC18/19 plasmid contains:
• The pMB1 repliconrep responsible for plasmid replication (Source – PlasmidpBR322). The high copy number of the pUC plasmid is due to the lack of therop gene and a single point mutation in the pMB1 repliconrep. The
•bla gene (encoding beta-lactamase) confers resistance to ampicillin (source – plasmid pBR322). It differs from pBR322 in two point mutations
• E. coli lac operator region, including the CAP protein binding site, promoter P_lac, < The i>lac repressor binding site and the 5' terminal portion of the lacZ gene encoding the N-terminal fragment of β-galactosidase (source – M13mp18/19). This fragment is inducibly synthesized by IPTG and is capable of intra-allelic (α) complementation utilizing a defective form of β-galactosidase encoded by the host (mutated delta(lacZ)M15). In the presence of IPTG, the bacteria synthesize these two fragments of the enzyme and form blue colonies on medium containing X-Gal. DNA insertion into the MCS located within the lacZ gene (codons 6-7 of lacZ are replaced by MCS) can lead to the inactivation of the N-terminal fragment of β-galactosidase and Eliminate alpha complementarity. Therefore, bacteria carrying the recombinant plasmid will produce white colonies

The diagram shows the enzyme that performs one cleavage of pUC18 DNA. Enzymes produced by Thermo Scientific are shown in orange. Coordinates refer to the position of the first nucleotide in each recognition sequence.

The exact location of the genetic element (including the stop codon) is shown in the map. bla gene nucleotides 2486-2418 (complementary strand) encode a signal peptide. The LacZ polypeptide corresponding to wt β-galactosidase and required for blue-white screening terminates at nt position 236 (complementary strand). Another 30 codon in the same reading frame is derived from pBR322. The indicatedrep regions are sufficient to facilitate replication. DNA replication is initiated at position 866 (± 1) and proceeds in the direction indicated. Plasmids carrying the pMB1 and ColE1 replicons are incompatible, but they are fully compatible with plasmids carrying the p15A replicon (pACYC177, pACYC184). The pMB1-derived plasmid can be amplified using chloramphenicol.

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