The insert contains the following sites (with nucleotide positions according to the published sequence): NaeI at -426; HindIII at -420; SacI at -408 and PstI at -307. Restriction digests of the clone give the following sizes (kb): EcoRI--3.0, 0.8; HindIII--3.65, 0.15; BamHI--3.8; PstI--3.8; XbaI/SacI--2.9, 0.7, 0.2. The 5' clone phgJE3-3 (ATCC 61366/61367) and the 3' clone phgJE2-1 (ATCC 61368/61369) together generate a complete gene. This insert was subcloned from lambdahJE-7 and extends from -531 relative to the mRNA cap site into the first intron which begins as +228. It is oriented in this clone with transcription from the SP6 promoter giving the + strand. |