lambdaMGU2 (ATCC® 77367)

Clone Type: Clone  /  Depositors: IN Maruyama

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Designations lambdaMGU2
Depositors IN Maruyama
Vector for constructing cDNA libraries
Vector permitting positive selection for inserts
Vector permitting production of single-stranded DNA
Vector Vector type: phage
Vector size (kb): 41.7
Markers: ampR
Replicons: lambda, pMB1, M13
Cloning site: BamHI
Suggested host: E. coli Q358
Related Products
Component of: ATCC 77369
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Shipping Information Freeze-dried bacteria-free lysate
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.

Efficiency of phagemid recovery is approximately 20%. Plasmid pCRE1 may be a low level contaminant, but is easily distinguished from pMGU DNA.

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.

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.

To enable the positive selection of inserts, the library should be plated on a P2 lysogen such as Escherichia coli Q359 (ATCC 47019).

Inserts can be amplified using the following primers flanking the BamHI cloning site: upstream 5'-AAGAGGCAGAACTGGCAG-3' and downstream 5'-ATCGATGCATAGCGATTC-3'.

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.

Maruyama IN, Brenner S. A selective lambda phage cloning vector with automatic excision of the insert in a plasmid. Gene 120: 135-141, 1992. PubMed: 1327972