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

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.

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

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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