OmniGene, Inc., P Mahler, OmniGene, Inc.
Size (kb): 14.8000001907348600
Vector: pKB1020 (plasmid)
Promoters: Promoter lambda PL
Construction: pEG5005, lambda cassette
Construct size (kb): 14.80000019073486
Features: marker(s): ampR
promoter: lambda PL
repressor gene: Mu cts62
repressor gene: cI857
restriction site: KpnI
restriction site: SstII
coding sequence: A
coding sequence: B
coding sequence: N
coding sequence: int
coding sequence: xis
contains sequence transcription antitermination (N) protein
Restriction digests of the clone give the following sizes (kb): ClaI--14.5, 0.3; KpnI--14.8; BglII/KpnI--11.0, 3.5, 0.3.
To maintain the plasmid, grow at 30C with ampicillin and kanamycin selection.
Thermal induction (at 42C) of the Mu cts lysogen containing the plasmid yields a low titer phage lysate. Lysates should be used within 1 week of preparation because the clones can be unstable.
Infection of sensitive, Mu cts lysogenic E. coli strains and growth at 30C yields two types of kanamycin-resistant isolates: ampicillin-sensitive strains (integrants) that carry no plasmid DNA, and ampicillin-resistant strains that contain the plasmid.
Host integration vector for adapting E. coli strains as hosts for att-Int mediated rearrangement and regulation of lambda-based expression vectors.
Constructed by insertion of a lambda gene cassette into the bacteriophage Mu based vector pEG5005 (ATCC 67463)
, which is defective for the morphogenic functions.
Balakrishnan R, et al. A gene cassette for adapting Escherichia coli strains as hosts for att-Int-mediated reqrrangement and pL expression vectors. Gene 138: 101-104, 1994. PubMed: 8125284
Groisman EA, Casadaban MJ. Mini-Mu bacteriophage with plasmid replicons for in vivo cloning and lac gene fusing. J. Bacteriol. 168: 357-364, 1986. PubMed: 3020001