Restriction digests of the clone give the following sizes (kb): ClaI--9.3; HindIII--4.9, 2.55, 1.85; XbaI--uncut; NotI--9.3. YI-type shuttle integrative vector targeting human LINE sequences in YAC libraries derived from somatic cell hybrids. Constructed by replacing a 2.2 kb XbaI fragment of the LINE element in pL1.1a with a blunt-ended 1.2 kb fragment containing HIS3. The replaced element, nt 3392 to 5607 of the LINE consensus sequence, contained conserved sequences which might have permitted targeting to murine-derived YACs. Linearizing with NotI and ClaI removes plasmid sequences from the LINE ends for targeting integration. The NotI site is at the 5' end of the LINE sequence. |
