pBAD30 (ATCC® 87401)

Applications: expression vectorproduces protein arabinose regulatorvector containing primer sites useful for sequencingvector with low copy number  /  Depositors: L Guzman, J Beckwith

Designations pBAD30
Depositors L Guzman, J Beckwith
Biosafety Level 1
Vector Information
Size (kb): 4.923
Vector: pBAD30 (phagemid)
Promoters: Promoter araC
Construction: pBAD28
Construct size (kb): 4.923
Features: marker(s): ampR
marker(s): cmlR
operator: I2 + I1
operator: O1
operator: O2
other: CAP site
promoter: araC
promoter for expression: arabinose BAD
regulator: araC
replicon: M13
replicon: p15A
MCS: NheI...HindIII
transcription terminator: rrnB T1 + T2
expression vector
produces protein arabinose regulator
vector containing primer sites useful for sequencing
vector with low copy number
Restriction digests of the clone give the following sizes (kb): AvaI--2.0, 1.85, 1.2; BamHI--5.0; PstI--4.0, 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'.
Plasmid copy number is low due to the p15A replicon. This vector can be used when reduced gene expression is desirable.
Plasmid contains the 3' portion of the cat (cmlR) gene which could promoter recombination if this plasmid is used in combination with other compatible cmlR plasmids.
Plasmid is compatible with pBR-derived plasmids and may be used for coexpression of cloned inserts.
Recombination can be avoided by the use of recA host strains, or it can be used to advantage to intentionally exchange markers among plasmids.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C

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

Shipped frozen
Shipping Information

 Frozen glycerol stock of E. coli containing the plasmid.