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Construct size (kb): 8.100000381469727

DESCRIPTION OF VECTOR COMPONENT:
Name of vector: pHSS21
Intact vector size:
Type of vector: plasmid
Vector end: EcoRI
Vector end: XhoI
Cloning sites: HindIII SphI SalI XbaI BamHI XhoI KpnI SacI EcoRI
Polylinker sites: HindIII SphI PstI SalI XbaI BamHI XhoI KpnI SacI EcoRI
Construction: pUC19
Host range: Escherichia coli
Features (with orientation and position when available):
marker(s): kanR
promoter: lac
replicon: pMB1
Cross references:

DNA: genomic

DESCRIPTION OF INSERT COMPONENT:
Genome: Bacillus subtilis
Gene symbol: sacB
Gene name: levansucrase
Contains complete coding sequence?: U
Genome: Bacillus subtilis
Gene symbol: sacR
Gene name: sucrase, levansucrase regulator
Contains complete coding sequence?: U
Genome: pCU1
Gene symbol: oriT
Gene name: (IncN) origin of transfer replication
Contains complete coding sequence?: U
Type of DNA: genomic
Insert end: EcoRI
Insert end: SalI
Insert size (kb): 5.0
Cross references:

Insert lengths(kb): 5.0

Gene product: (IncN) origin of transfer replication [oriT]

Restriction digests of the clone give the following sizes (kb): NotI--5.6, 2.4; EcoRI/HindIII--2.8, 1.5, 1.15, 0.9, 0.7, 0.55, 0.2; HindIII--4.2, 1.8, 0.9, 0.8.

Available for research purposes only.

Expression of sacB confers sensitivity to sucrose (no growth on LB medium containing 5% sucrose).

Plasmid containing an oriT/cmlR/sacBR mobilization cassette that can be inserted into a general purpose vector (ATCC 77278, 77279) for conjugal transfer of cloned inserts into the chromosome of P. aeruginosa.

Constructed by inserting a 0.9 kb BamHI fragment containing cmlR into the BamHI site of pMOB2 (ATCC 77281). The NotI digest excises a fragment of approximately 5.4 kb that includes short segments of the pHSS21 sequence.

The NotI cassette includes these features: pHSS21 sequence - EcoRI - oriT - PstI - BamHI - HindIII - cmlR - HindIII - BamHI - XbaI - SalI - PstI - sacBR/HindIII/EcoRI/XbaI/SalI - PstI - SphI - HindIII - pHSS21 sequence.