Size (kb): 12.6999998092651400
Vector: pJEL144 (plasmid)
Promoters: Promoter deoP2
Construction: pJEL170, pJEL132
Construct size (kb): 12.69999980926514
Features: insert detection: lacZ'
repressor gene: cI857
vector with low copy number
Restriction digests of the clone give the following sizes (kb): BamHI--13.6; EcoRI--13.6.
Transcription from deoP2 is negatively regulated by the DeoR and CytR repressors and depends on the cAMP/CRP complex.
Below 37C, there is one copy per genome equivalent stabilized by the parB locus. At 42C replication is uncontrolled because induction of the cI857-regulated PR promoter permits expression of the copB, copA, and repA genes.
Expression of beta-galactosidase from this vector can be increased by the use of hosts containing repressor mutations such as S0929 (cytR), S0930 (deoR) and S0931 (cytR, deoR).
Terminator-testing plasmid. Termination signal-containing sequences cloned into the BamHI site may reduce expression of beta-galactosidase activity regulated by the deoP2 promoter.
Constructed by inserting a 0.22 kb EcoRI/BamHI fragment containing the deoP2 regulatory region and the first 16 codons of deoC from pJEL134 into pJEL170.
Larsen JE, et al. Analysis of the terminator region after the deoCABD operon of Escherichia coli K-12 a new class of single copy number operon-fusion vectors. Nucleic Acids Res. 15: 5125-5140, 1987. PubMed: 3299264