pBAD18 (ATCC® 87393)

Applications: expression vector  produces protein arabinose  regulator vector containing primer sites useful for sequencing  /  Depositors: L Guzman, J Beckwith

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Designations pBAD18
Depositors L Guzman, J Beckwith
Biosafety Level 1
Vector Information
Size (kb): 4.613
Vector: pBAD18 (phagemid)
Promoters: Promoter araC
Construction: pBAD18sc
Marker(s): ampR,araC
Construct size (kb): 4.613
Features: marker(s): ampR
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: pMB1
MCS: NheI...HindIII
transcription terminator: rrnB T1 + T2
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'.
Expression of cloned inserts in the two recommended hosts differ in terms of induction characteristics, especially when using low concentrations of inducer (arabinose) and growing in minimal media.
Cloned inserts must 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°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 containing the plasmid
Product Sheet
Product Sheet
Product Sheet
Attachments Vector Map