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Host
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Escherichia coli HB101 (ATCC 33694)
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Vector Information
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Intact vector size: 5.900
Type of vector: plasmid
Cloning sites: EcoRI SacI KpnI SmaI BamHI XbaI SalI AccI PstI SphI HindIII
Polylinker sites: EcoRI SacI KpnI SmaI BamHI XbaI SalI AccI PstI SphI HindIII
Construction: pUB110, pBR322, vegII
Host range: Bacillus subtilis; Escherichia coli
Features (with orientation and position when available):
terminator: To phage lambda
promoter: vegII (B. subtilis)
MCS: HindIII...EcoRI
transcription terminator: rrnB
marker(s): ampR
replicon: pMB1
replicon: pUB110
marker(s): neoR
marker(s): bleR
Vector: pRB374 (plasmid)
Promoters: Promoter vegII (B. subtilis)
Construction: pUB110, pBR322, vegII
Marker(s): ampR,bleR,neoR
Construct size (kb): 5.900
Features: marker(s): ampR
marker(s): bleR
marker(s): neoR
promoter: vegII (B. subtilis)
replicon: pMB1
replicon: pUB110
MCS: HindIII...EcoRI
terminator: To phage lambda
transcription terminator: rrnB
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Applications
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expression vector shuttle vector
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Comments
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Restriction digests of the clone give the following sizes (kb): EcoRI--5.9; BamHI--5.9; BglI/BglII--3.5, 2.4. The Bacillus subtilis promoter vegII initiates transcription in both B. subtilis and E. coli. Structural stability of the plasmid in B. subtilis can be affected by high levels of protein production. Under these conditions, cell growth and stability may be improved by reducing the antibiotic concentration in the media. May not be suitable for cloning very strong expression signals. Neo confers resistance to neomycin and kanamycin and ble confers resistance to bleomycin and phleomycin.
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References
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Bruckner R. A series of shuttle vectors for Bacillus subtilis and Eschericia coli. Gene 122: 187-192, 1992. PubMed: 1452028
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