Description

pUC118/119 DNA
pUC118/119 are plasmid vectors developed for the preparation of single stranded DNA. pUC118/119 was constructed by inserting intergenic region (IG region) of the M13 phage DNA into the pUC18/pUC19 plasmid.
Therefore, infection by the helper phage M13K07 induces the production of pUC118/pUC119 as single stranded DNA which is predominantly packaged into phage particles and is then released from bacterial cells. In addition, there is almost no contamination by the single stranded DNA of the helper phage. Using this system, single stranded DNA from large DNA fragments (up to 7 kb) can be stably obtained without deletion. pUC118 and pUC18 contains the consensus multi-cloning sites, samely pUC119 and pUC19 also.

pUC118 DNA (BAP treated)
pUC118 DNA (BAP-treated) is a useful cloning vector, prepared by cleavage with a frequent restriction endonuclease which is cleavable only single site in multicloning sites of pUC118, then dephosphorylated by alkaline phosphatase purified from E. coli (BAP). This DNA is able to prevent self-ligation of vectors, and also able to decrease blank value of transforming cells. Thus, this DNA is available to cloning without other treatment.

pTV118N DNA
pTV118N DNA is plasmid vector (phagemids vector) constructed by modifying pUC118 for the preparation of single stranded DNA like pUC118/119. This plasmid codes lacZ α peptide and is constructed by inserting Nco I cleavage sequence (CCATGG) into the initiation codon (ATG) site of pCU118 plasmid. Therefore, this plasmid induces the expressoin of foreign gene which is inserted into the Nco I site by utilizing lac promoter, lacZ SD sequence, and the initiation codon and by adjusting the translation frame. The number of bases between SD sequence and the initiation codon is 8, which allows high level expression. In addition, recovery of single-stranded DNA by using helper phage and the subsequent sequencing with RV-N primer enables to read the sequence from the initiation codon site and the translation frame can be confirmed easily.

Note:

The single-stranded DNA obtained from pTV118N DNA can not be sequenced with M13 forward primer since the direction of M13 IG region for lacZ of pTV118 is opposite to of pUC118.

Storage

–20°C

Concentration

pUC118, pUC119, pTV118N	:	250-1,000 µg/ml
pUC118 DNA (BAP treated)	:	50-250 µg/ml

Chain length (Calculated with Messing method)

pUC118/119	:	3,162 bp
pTV118N	:	3,163 bp

Purity

·	Double-strand covalently closed circular form I (RF I) comprises more than 70%.
·	It is confirmed that single-strand DNA is secreted upon infection of helper phage (M13KO7).
·	It is confirmed that the multi-cloning site is well-maintained upon sequence analysis by dideoxy method.
pUC118/119 DNA
·	It is confirmed that the vector contains a single cleavage site for EcoR I, Sac I, Kpn I, Sma I, BamH I, Xba I, Sal I, Pst I, Sph I and Hind III upon restriction analysis by the enzymes.
pUC118 DNA (BAP treated)
·	The following were confirmed for pUC118 (BAP treated); This product shows single band on 1% agarose gel electrophoresis. When transformation was performed by this product after treated with T4 DNA ligase, back ground transformants was detected below 1% compared with the case of using uncleaved pUC118. When this product was used on transformation of E. coli JM109 competent cells (1 × 107 transformants/1 µg pUC118) after treatment with polynucleotide kinase and subsequent treatment with T4 DNA ligase, it was shown that efficiency of was above 1 × 106 transformants/1 µg DNA. White colonies were detected below 1% in addition of IPTG and X-Gal.
pTV118N DNA
·	It is confirmed that the vector contains a single cleavage site for EcoR I, Sac I, Kpn I, Sma I, BamH I, Xba I, Sal I, Pst I, Sph I, Hind III and Nco I upon restriction analysis by the enzymes.

GenBank

	Entry Name	Accession No.
pUC118	CVU07649	U07649
pUC119	CVU07650	U07650

Intended usage

• Cloning of a target gene
• DNA sequencing by using M13 primers
• Determination of long DNA sequences using Deletion Kit for Kilo-Sequencing (Cat.# 6030)
• Gene expression using lac promoter

Form

10 mM Tris-HCI (pH8.0), 1 mM EDTA

The sequence of multi-cloning site is the same between pUC118/119 and pUC18/19.
pTV118N has the same sequence in multi-cloning as pUC18.
Plasmid vector map of pUC118/119, pTV118N

Notice

The single-stranded DNA obtained from pTV118N DNA can not be sequenced with M13 forward primer since the direction of M13 IG region for lacZ of pTV118 is opposite to of pUC118.