Restriction digests of the clone give the following sizes (kb): BglI--1.75 (doublet); EcoRI--3.5; PstI--2.1, 1.45. Restriction digests of the plasmid give the following sizes (kb): BamHI--3.507; BglI--1.784, 1.723; PstI--2.072, 1.435; PvuI--2.063, 1.444; PvuII--2.852, 0.655; SacI--3.257, 0.25. Inserts can be cloned into the vector using a filled-in NcoI site and a 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 producing proteins with the one additional N-terminal methionine. A modified lacZ' allows blue/white screening of colonies. Constructed from pfXblue (ATCC 77333) by deletion of lambda cII and coagulation factor Xa protease recognition sequences (nt 824-943). The order of the major features of the plasmid is: pMB1 ori - ampR - lambdaPL promoter - NcoI/MCS/HindIII/modified lacZ'. |
