Restriction digests of the clone give the following sizes (kb): BglI--1.8 (doublet); EcoRI--3.6; PstI--2.1, 1.55. Restriction digests of the plasmid give the following sizes (kb): BamHI--3.574, 0.053; BglI--1.843, 1.784; PstI--2.072, 1.555; PvuI--2.063, 1.564; PvuII--2.972, 0.655; SacI--3.257, 0.37. Inserts can be cloned using the 5' StuI site and one other MCS restriction site matching the 3' end of the insert. The vector supplies the AUG initiator codon. The insert to be cloned should have a phosphorylated 5' blunt end with the first base corresponding to the first base of the first codon. Expression vector permitting the production of a fusion protein that can be trimmed by cleavage with the coagulation factor Xa protease. A modified lacZ' allows blue/white screening of colonies. Especially useful for the expression of PCR products. The order of the major features of the plasmid is: pMB1 ori - ampR - lambdaPL promoter - lambda cII - factor Xa recognition sequence - StuI/MCS/HindIII/modified lacZ'. |
