pEX100T (ATCC® 87436)

Applications: vector permitting positive selection for integration levansucrasevector permitting visual detection of recombinants  /  Depositors: HP Schweizer

Designations pEX100T
Depositors HP Schweizer
Biosafety Level 1
Vector Information
Size (kb): 5.8460001945495610
Vector: pEX100T (plasmid)
Construction: pUC19Sce, pEM5, pMOB2
Marker(s):ampR,sacB
Construct size (kb): 5.846000194549561
Features: marker(s): ampR
marker(s): sacB
replicon: oriT
replicon: pMB1
restriction site: I-SceI
restriction site: SmaI
transcription terminator: rrnB T1
transcription terminator: rrnB T2
coding sequence: lacZalpha
Applications
vector permitting positive selection for integration levansucrase
vector permitting visual detection of recombinants
Comments
Restriction digests of the clone give the following sizes (kb): ClaI--5.6, 0.40; ScaI--6.0; SmaI--6.0; AvaI--6.0.
The ClaI restriction recognition site located at bp 2093 is subject to dam methylation (gATCGAT) in its distribution host DH5alphaF'.
The ampR marker can easily be replaced by other antibiotic resistance-encoding cassettes by cloning into the unique ScaI site contained within the bla gene.
E. coli strains containing sacB are sucrose sensitive on LB medium + 5% sucrose. It is important to monitor sucrose sensitivity at each cloning step, as spontaneous mutations can accumulate during the process which then confer sucrose resistance.
Vector to construct mutants in Pseudomonas aeruginosa.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Schweizer HP, Hoang TT. An improved system for gene replacement and xylE fusion analysis in Pseudomonas aeruginosa. Gene 158: 15-22, 1995. PubMed: 7789804

Shipped frozen
Shipping Information

 Frozen glycerol stock of E. coli containing the plasmid

Product Sheet
Product Sheet
Product Sheet
Attachments Vector map for 87436