Size (kb): 6.0260000228881840
Vector: pBAD18-Cm (phagemid)
Promoters: Promoter araC
Construction: pBAD18, cmlR (pACYC184)
Construct size (kb): 6.026000022888184
Features: marker(s): cmlR
operator: I2 + I1
other: CAP site
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--3.8, 2.3; HindIII--6.0; PstI--5.0, 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'.
Cloned inserts must provide a translation initiation sequence (ATG) and ribosome binding site for expression.
Plasmid contains bla (ampR) sequences surrounding the cmlR gene which could promoter recombination if this plasmid is used in combination with other compatible ampR plasmids.
Recombination can be avoided by the use of recA host strains, or it can be used to advantage to intentionally exchange markers among plasmids.
Freeze dried Escherichia coli containing the plasmid.