Size (kb): 4.613
Vector: pBAD18 (phagemid)
Promoters: Promoter araC
Construct size (kb): 4.613
Features: marker(s): ampR
operator: I2 + I1
other: CAP site
promoter: bla (ampR)
promoter for expression: arabinose BAD
transcription terminator: rrnB T1 + T2
produces protein arabinose regulator
vector containing primer sites useful for sequencing
Restriction digests of the clone give the following sizes (kb): EcoRI--4.6; HindIII--4.6; PstI--3.6, 1.1.
Cultures should be grown in minimal media for more reproducible induction of expression. Expression is induced in glycerol-containing media by addition of arabinose. Expression is repressed by addition of glucose or other catabolites.
One of several tightly controlled expression vectors (ATCC 87393-87402) regulated by the arabinose operon. The vectors differ in replicon, antibiotic resistance marker, multiple cloning site and mechanism of initiation of translation.
The following primers can be used for sequencing of cloned inserts: 5' primer (27 - 8 bp upstream of the NheI site) 5'-CTGTTTCTCCATACCCGTT-3'; and one of two 3' primers: 3' primer 1 (2 - 19 bp downstream of the HindIII site) 5'-CTCATCCGCCAAAACAG-3';
3' primer 2 (17 - 33 bp downstream of the HindIII site) 5'-GGCTGAAAATCTTCTCT-3'.
Expression of cloned inserts in the two recommended hosts differ in terms of induction characteristics, especially when using low concentrations of inducer (arabinose) and growing in minimal media.
Cloned inserts must provide a translation initiation sequence (ATG) and ribosome binding site for expression.
Guzman LM, et al. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter. J. Bacteriol. 177: 4121-4130, 1995. PubMed: 7608087
Frozen glycerol stock of E. coli containing the plasmid