This vector contains the following restriction sites (in approximate kb from one PstI site): KpnI--0.8, 3.8; BamHI--4.4, 10.7, 13.4; HpaI--5.8; EcoRI--8.3; SalI--10.3; XhoI--12.5. The original vials from the depositor are being used for distribution. The pGA22 portion of the construct contains the drug resistance genes. cmlR can be insertionally inactivated by cloning into the EcoRI site; kanR can be inactivated by cloning into the HindIII or XhoI sites. Growth at 30C in the presence of chloramphenicol, kanamycin, or tetracycline was verified. Recommended tetracycline concentration is 1.5 ug/ml on LB agar. This was constructed by ligating pCG2 (6.6 kb) linearized with PstI to pGA22 (7.9 kb) linearized with PstI. |
