Vector Database

Deposited by: Walter Messer Vector containing the attP sequence for integrating DNA into the lambda attachment site attB. Requires the lambda integrase (int) such as that provided by pLDR8 (ATCC 77357). [1] When a recombinant construct is digested with NotI, the origin is separated from the insert. Each fragment has a distinct selectable marker. After religation to form origin-free circles, the products are transformed into bacteria with pLDR8. [1] Expression of int in pLDR8 is regulated by lambda PR, by the temperature sensitive cI857 repressor. Replication of pLDR8 is also temperature sensitive. [1] Therefore, at 30C replication of the plasmid is normal and int is not expressed. At 42C, int is expressed but pLDR8 is no longer replicated, resulting in loss of int after a few cell generations. [1] The order of the major features in this plasmid is: NotI - ampR - ClaI/MCS/EcoRI - attP - NotI - pMB1 ori - cmlR. [1] Restriction digests of the clone give the following sizes (kb): EcoRI/BglII--1.8, 1.6, 0.5; BamHI--3.9; XbaI--3.9; NotI--2.0, 1.9. (ATCC staff) Medium is 1227 LB plus ampicillin. E.coli LJ24 for integration Hosts: E.coli, E.coli WM1202, E.coli JM103, E.coli JM109, E.coli LJ24. (Information source: VectorDB ( http://seq.yeastgenome.org/vectordb ).)