pBAD33 (ATCC® 87402)

Applications: Expression vector Produces protein arabinose regulator Vector containing primer sites useful for sequencing Vector with low copy number  /  Depositors: L Guzman, J Beckwith

Designations pBAD33
Depositors L Guzman, J Beckwith
Biosafety Level 1
Host Escherichia coli DH5alpha
Vector Information
Intact vector size: 5.356
Type of vector: phagemid
Cloning sites: SacI KpnI SmaI XbaI SalI PstI SphI HindIII
Polylinker sites: NheI EcoRI SacI KpnI SmaI BamHI XbaI SalI AccI PstI SphI HindIII
Construction: pBAD28
Host range: Escherichia coli
Features (with orientation and position when available):
regulator: araC, <-, 3452-4330
operator: O2, 4359-4376
promoter: araC, <-, 4481-4509
operator: O1, 4517-4538
other: CAP site, 4560-4573
operator: I2 + I1, 4569-4607
promoter for expression: arabinose BAD, ->, 4606-4633
MCS: NheI...HindIII, ->, 4656-4718
transcription terminator: rrnB T1 + T2, ->, 4719-5144
replicon: M13, 335-793
marker(s): cmLR, <-, 1348-2007
replicon: p15A, 2369-3213
Marker(s): cmlR,araC
Applications
Expression vector
Produces protein arabinose regulator
Vector containing primer sites useful for sequencing
Vector with low copy number
Comments
Restriction digests of the clone give the following sizes (kb): AvaI--3.0, 1.3, 1.2; BamHI--5.4; EcoRI--2.95, 2.6.
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 bla (ampR) sequences following the rrnB terminator which could promoter recombination if this plasmid is used in combination with other compatible ampR 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 1675: LB medium (ATCC medium 1065) with 10 mcg/ml Chloramphenicol
Growth Conditions
Temperature: 37°C
References

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
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