Restriction digests of the clone give the following sizes (kb): NotI--5.5; BglII--5.5; SalI--5.5; EcoRI--5.5; SacI--5.5. pJS97 (ATCC 77153) and pJS98 (ATCC 77154) are used together to construct artificial chromosomes. Each plasmid is digested with ClaI and the cloning enzyme. To recover both ends of a YAC insert as plasmid, DNA of a YAC-containing strain is digested to completion with BamHI, SphI or SacI, diluted, ligated, and transformed into E. coli (to ampicillin resistance). The insert of a resultant plasmid (which includes the T7 promoter) extends from an end of the YAC insert to the first site of the enzyme used for digestion. YL-type shuttle vector useful for construction of genomic libraries. Contains promoters for in vitro RNA synthesis and permits recovery of end fragments as plasmids in E. coli. The order of the major features in this plasmid is: ClaI/MCS/EcoRI - T7 promoter - ARSH4 - TRP1 - pMB1 ori - bla - SacI - KpnI - BamHI - SphI - TEL. |
