pJEL144 (ATCC® 37526)

Applications: terminator-cloning vectorvector with low copy number  /  Depositors: P Valentin-Hansen

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Designations pJEL144
Depositors P Valentin-Hansen
Biosafety Level 1
Vector Information
Size (kb): 12.6999998092651400
Vector: pJEL144 (plasmid)
Promoters: Promoter deoP2
Construction: pJEL170, pJEL132
Marker(s):ampR
Construct size (kb): 12.69999980926514
Features: insert detection: lacZ'
marker(s): ampR
promoter: deoP2
replicon: R1
repressor gene: cI857
terminator: none
Applications
terminator-cloning vector
vector with low copy number
Comments
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.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

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

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