The insert contains the following restriction sites (nt from the 5' end): RsaI--323; BamHI--420. Restriction digests of the clone give the following sizes (kb): HindIII--4.6; EcoRI--4.6; AvaI--4.6; BamHI--3.5, 1.05; PstI--3.9, 0.68. The insert extends from the second nucleotide of the codon for amino acid -15 of the signal peptide to the poly(A) tract. Detects genomic EcoRI fragments of 8.0, 6.6, and 4.3 kb in human cell lines, and fragments of 6.7, 2.1, and 1.9 kb in mouse. This probe also reveals a HindIII polymorphism which appears to be linked to the PvuII polymorphism. The vector pKT218 is pBR322 with an approximate 600 bp deletion of the ampR gene. The deletion starts near the beginning of the amp gene but is distal to the amp promoter and initiation ATG site and ends at the PstI site. The sequence from the ATG start of the amp gene to the PstI site is :
ATG AGT ATT CAA GCT GCA G This construct comes as 200 ng of purified plasmid DNA. It is also available already in E. coli as ATCC# 59944. |
