pBP103

Generic chromosome fragmentation vector containing promoters for in vitro RNA synthesis. The unique cloning sites can be used to insert targeting sequences, such as cDNAs. [1] Any unique restriction site between the targeting sequence and the telomere-adjacent sequence can be used to linearize the plasmid before transformation. [1] The EcoRV and ClaI sites between the SP6 promoter and the telomere sequence can be used for recovery from deletion derivatives of YAC insert sequences adjacent to the fragmentation site. [1] A 0.24 kb EcoRI/SalI fragment containing the telomere-adjacent sequence from YTCA-1x was ligated into the MCS of pSP73. The EcoRI and BamHI sites of the MCS were destroyed and a 0.3 kb NdeI/PvuII fragment was inserted into the PvuII site of pRS303. [1] Restriction digests of the clone give the following sizes (kb): BamHI–4.6; EcoRI–4.6; SalI–4.6; SalI/EcoRI–4.4, 0.3. (ATCC staff) The order of the major features in this plasmid is: bla – HIS3 – lacZ’/MCS – HindIII – SphI – PstI – AccI – SalI – TEL – ClaI – EcoRV – SP6 promoter. [1] Medium is 1227 LB plus ampicillin. Hosts: E.coli HB101, Saccharomyces cerevisiae, E.coli, Saccharomyces cerevisiae YPH252, Saccharomyces cerevisiae CGY2516 [ATCC 74013].