Restriction digests of the clone give the following sizes (kb): ClaI--4.3 (doublet); EcoRI--7.0, 1.6; SacI--4.6, 4.0; BamHI--4.7, 2.8, 1.2. Acentric chromosome fragmentation vector targeting LINE sequences. Contains promoters for in vitro RNA synthesis. Any unique restriction site between the targeting sequence and the telomere-adjacent sequence can be used to linearize the plasmid before transformation. 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. Constructed from pBP103 (ATCC 77087) by inserting a 4.0 kb LINE-containing SacI fragment from pL1.1A into the SacI site. pBP110 (ATCC 77090) and pBP111 (ATCC 77091) differ in the orientation of the LINE sequence. The order of the major features in this plasmid is: bla - HIS3 - lacZ'/MCS/3'LINE5' - HindIII - SphI - PstI - AccI - SalI - TEL - ClaI - EcoRV - SP6 promoter. |
