Vector Database

Deposited by: Ichiro N. Maruyama Restriction digests of the clone give the following sizes (kb): HindIII--24.1, 17.6; EcoRI--22.0, 20.0; BamHI--24.8, 16.9; XbaI--32.7,9.0; NotI--41.7. (ATCC staff) Vector useful for constructing cDNA libraries. Permits positive selection for inserts using the Spi- phenotype, and excision of phagemid by lox/cre site-specific recombination. [1] To prepare phagemid from lambdaMGU2, grow recombinants on a RecA- host expressing the Cre protein (E. coli 1046[pCRE1], ATCC 77368) and select for ampicillin resistance. The pMGU product is 4.185 kb. [1] The order of the major features in the cloning region of the lambda vector is: lambda J - SmaI - SalI - loxP - EcoRI - M13 ori - ampR - pMB1 ori - HindIII - 3'gam/BamHI/5'gam - XhoI - loxP - SalI - lambda N. [1] Inserts can be amplified using the following primers flanking the BamHI cloning site: upstream 5'-AAGAGGCAGAACTGGCAG-3' and downstream 5'-ATCGATGCATAGCGATTC-3'. [1] Efficiency of phagemid recovery is approximately 20%. Plasmid pCRE1 may be a low level contaminant, but is easily distinguished from pMGU DNA.[1] To enable the positive selection of inserts, the library should be plated on a P2 lysogen such as Escherichia coli Q359 (ATCC 47019). [1] Medium is 1592 SM buffer. EcoRI-HindIII fragment is 1039 bp. This sequence comes from Fig. 3. [1] NCBI gi: 258428 Hosts: E.coli, E.coli Q358, bacteria-free lysate. (Information source: VectorDB ( http://seq.yeastgenome.org/vectordb ).)