pBAD22 (ATCC® 87398)

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

Permits and Restrictions

View Permits

Designations pBAD22
Depositors L Guzman, J Beckwith
Biosafety Level 1
Vector Information
Size (kb): 4.5419998168945310
Vector: pBAD22 (phagemid)
Promoters: Promoter araC
Construction: pBAD18
Marker(s):ampR,araC
Construct size (kb): 4.541999816894531
Features: marker(s): ampR
operator: I2 + I1
operator: O1
operator: O2
other: CAP site
other: Kozak sequence
promoter: araC
promoter: bla (ampR)
promoter for expression: arabinose BAD
regulator: araC
replicon: M13
replicon: pMB1
MCS: NheI...HindIII
ribosome-binding site: Shine-Dalgarno sequence
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--4.4; BamHI--4.3, 0.2; AvaI--1.8, 1.65, 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'.
Vector contains an optimized Shine-Dalgarno ribosome binding site, translation initiation codon and Kozak sequence (relevant for expression in eukaryotic cells) which promote a high level of expression of cloned inserts.
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 frozen
Product Sheet
Product Sheet
Product Sheet
Attachments Vector Map