pBAD18s (ATCC® 87394)

Applications: expression vectorproduces protein arabinose regulatorvector containing primer sites useful for sequencing  /  Depositors: L Guzman, J Beckwith

Permits and Restrictions

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Designations pBAD18s
Depositors L Guzman, J Beckwith
Biosafety Level 1
Vector Information
Size (kb): 4.6360001564025880
Vector: pBAD18s (phagemid)
Promoters: Promoter araC
Construction: pBAD18sc
Marker(s):ampR,araC
Construct size (kb): 4.636000156402588
Features: marker(s): ampR
operator: I2 + I1
operator: O1
operator: O2
other: CAP site
promoter: araC
promoter for expression: arabinose BAD
regulator: araC
replicon: M13
replicon: pMB1
MCS: EcoRI...HindIII
ribosome-binding site: Shine-Dalgarno sequence
transcription terminator: rrnB T1 + T2
coding sequence: 5' portion of araB
Applications
expression vector
produces protein arabinose regulator
vector containing primer sites useful for sequencing
Comments
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'.
Vector contains the Shine-Dalgarno box and 5 amino acids of the araB gene, but does not necessarily form fusion proteins with cloned inserts. Higher expression may still be achieved due to the second SD box despite out-of-frame cloning.
Cloned inserts should provide a translation initiation sequence (ATG) and ribosome binding site for expression.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°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 freeze-dried
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