pBAD28 (ATCC® 87400)

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

Designations pBAD28
Depositors L Guzman, J Beckwith
Biosafety Level 1
Vector Information
Vector Information:

Vector type: phagemid

Vector size (kb): 5.801

Markers: ampR, cmlR, araC

Polylinker sites: NheI, EcoRI, SacI, KpnI, SmaI, BamHI, XbaI, SalI, AccI, PstI, SphI, HindIII

Construction: pBAD18, pACYC184

Features: marker(s): ampR
marker(s): cmlR
operator: I2 + I1
operator: O1
operator: O2
other: CAP site
promoter: araC
promoter: bla (ampR)
promoter for expression: arabinose BAD
regulator: araC
replicon: M13
replicon: p15A
MCS: NheI...HindIII
transcription terminator: rrnB T1 + T2

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--3.1, 1.9, 1.3; BamHI--6.1; PstI--5.0;1.2.
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 is compatible with pBR-derived plasmids and may be used for coexpression of cloned inserts.
Media Medium 1949: LB medium (ATCC medium 1065) with 50 mcg/ml chloramphenicol and 100 mcg/ml ampicillin
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
Shipping Information Frozen glycerol stock of E. coli DH5-alpha; containing the plasmid.