pBAD18-Cm (ATCC® 87396)

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

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Designations pBAD18-Cm
Depositors L Guzman, J Beckwith
Biosafety Level 1
Vector Information
Size (kb): 6.0260000228881840
Vector: pBAD18-Cm (phagemid)
Promoters: Promoter araC
Construction: pBAD18, cmlR (pACYC184)
Marker(s):cmlR,araC
Construct size (kb): 6.026000022888184
Features: marker(s): cmlR
operator: I2 + I1
operator: O1
operator: O2
other: CAP site
promoter: araC
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--3.8, 2.3; HindIII--6.0; PstI--5.0, 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'.
Cloned inserts must provide a translation initiation sequence (ATG) and ribosome binding site for expression.
Plasmid contains bla (ampR) sequences surrounding the cmlR gene which could promoter recombination if this plasmid is used in combination with other compatible ampR plasmids.
Recombination can be avoided by the use of recA host strains, or it can be used to advantage to intentionally exchange markers among plasmids.
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
Shipping Information

 Freeze dried Escherichia coli containing the plasmid.

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