Size (kb): 8.0000000000000000
Vector: pQL1 (plasmid)
Construction: pACYC184, pKK232-8, kanR
Construct size (kb): 8.0
Features: marker(s): ampR
marker(s): kanR (neoR, rbR)
other: Haemophilus-specific uptake sequence
stop codons: 3 reading frames
transcription terminator: T1
transcription terminator: T1 T2
Restriction digests of the clone give the following sizes (kb): EcoRI--7.7, 0.3; BamHI--8.0; HindIII--8.0; PstI--3.5, 3.4, 0.9; SalI--4.6, 3.4.
Do not incubate with carbon dioxide when using ribostamycin.
The TSTE element is a 2.2 kb BamHI fragment that includes neoR from Tn5 flanked by Haemophilus-specific uptake sequences. Ribostamycin resistance is useful to select for the presence of neoR in both E. coli (40 ug/ml) and H. influenzae (15 ug/ml).
Copy number is estimated at 6-13 per cell in both H. influenzae and E. coli.
Protocol: 5% CO2
Heidecker GJ, et al. Construction of an ori cassette for adapting shuttle vectors for use in Haemophilus influenzae. Gene 150: 141-144, 1994. PubMed: 7959040
Terrence L Stull, personal communication