Description

This DNA is a plasmid vector suitable for DNA sequencing by dideoxy method. This DNA may allow successful cloning of DNA fragment longer than the case of M13 phage vector used. Since it has multi-cloning sites on lacZ' region, any DNA insertion into this vector can be easily verified using plates containing IPTG and X-Gal. Moreover, expression of the cloned DNA can be performed by using of lac promoter. M13 primer is available for DNA sequencing.
Using Deletion Kit for Kilo-Sequencing (Cat.# 6030), DNA sequencing of the cloned DNA with kilo-bases can also be performed.
Those are pUC type of cloning vector plasmid, pHSG298 and pHSG299 possessing kanamycin resistancy as selective markers, while pHSG396, pHSG397, pHSG398 and pHSG399 possessing chloramphenicol resistancy, respectively. pHSG298 and pHSG398 contain the consensus multi-cloning sites as same as pUC18, while pHSG299 and pHSG399 contain the consensus one as the same as pUC19 (even though Hind III and Sma I sites in pHSG298 and pHSG299 are not available). Multi-cloning sites of pHSG396 and pHSG397 are partially converted sites of pHSG399.

Storage

-20°C

Concentration

250-1,000 µg/ml

GenBank

	Entry Name	Accession No.
pHSG299	SYNHSG299	M19415

• Except the sequencing the multi-cloning site, pHSG298 has the same sequence at pHSG299. But pHSG298 lacks G at 1,446 in the sequence of pHSG299 registered with GenBank.
• The sequence of pHSG299 DNA (Cat.#3299) is different at the following position from the sequence of pHSG299 registered with GenBank.
  1663  C → deletion,  1808  C → T,  2228-2229  AT → TA,  2474  G → A,  2667-2668  GA → deletion

Chain length

pHSG298 : 2,675 bp
pHSG299 : 2,673 bp

Purity

• Double-strand covalently closed circular form I (RF I) comprises more than 70%.
• It is confirmed that the multi-cloning site is well-maintained upon sequence analysis by dideoxy method.

Intended usage

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

Form

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

Plasmid vector map of pHSG298/299