Our DNA cloning vectors may be used for a variety of cloning reactions, such as target gene cloning and expression, site-directed mutagenesis, and expression of genes that can be toxic to host cells. The suite of cloning vectors includes pKF18k-2, pKF19k-2, pSTV28, pSTV29, pTV118N, pTWV228, and pUC118.
pKF18k-2, PKF19k-2 DNA
The pKF pUC-derived vectors contain two amber stop mutations at the kanamycin resistance gene. When transformed with pKF, supE strains, like JM109, will grow on kanamycin-containing plates (sup0 strains, such as MV1184, will not). The pKF vectors can be used to perform site-directed mutagenesis based on the Oligonucleotide-directed Dual Amber (ODA) method. Moreover, pKF DNA is available for target gene cloning and expression via the lac promoter when transformed into supE hosts. Target genes cloned at the initiation codon (ATG) of the Nde I restriction site (CATATG) have a higher translation efficiency.
pSTV28, pSTV29 DNA
The pSTV vectors contain a beta-galactosidase gene, a pACYC184 origin of replication, a chloramphenicol resistance gene, and a pUC119 multiple cloning site. Since the pSTV plasmids produce fewer copy numbers compared with pUC-type high copy number plasmids, they are suitable for the expression of genes that may be toxic to host cells. The pSTV28 and pSTV29 vectors contain the pACYC184 origin of replication and can be co-transformed with pUC or pBR vectors.
The pTV118N DNA, a phagemid vector, is constructed from a modified pUC118 plasmid. pTV118N DNA contains the sequence CCATGG, which includes the cleavage sequence for the restriction enzyme NcoI as well as the start codon (ATG) for lacZ. This enables target gene expression at the NcoI site, while protein expression is enabled by the vector's lac promoter. The pTV118N vector also contains a lacZ SD sequence. There are eight bases between the lacZ SD sequence and the initiation codon, allowing high level expression of target genes. Induction of single-stranded DNA by helper phage and its subsequent sequencing with RV-N primer enables accurate sequencing from the start codon site, ensuring an insert's correct translation frame.
The pTWV228 vector contains a pBR322 origin of replication, an ampicillin-resistance gene, an intergenic region (IG region) of the M13 phage DNA, and a beta-galactosidase gene containing the multiple cloning site of pUC118. This vector is low-copy number, which makes it useful during the expression of genes that present potential toxicity to their host.
The pUC118 vectors can be used to prepare single-stranded DNA. pUC118 DNA was constructed by inserting the intergenic region (IG region) of the M13 phage DNA into a pUC18 plasmid. Co-transformation of pUC118 with the M13K07 helper phage induces the production of single-stranded DNA that is packaged into phage particles and released from bacterial cells. Using this system, large (up to 7 kb) and deletion-free single-stranded DNA can be stably obtained.
250–1000 µg/ml (except as specifically listed below)
pTV118N: 0.5 µg/µl
pUC118 BAP-treated DNA: 0.1 µg/µl
pKF 18k-2/19k-2 DNA: 0.5 µg/µl
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.