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Size (kb): 8.0000000000000000

Vector: pQL1 (plasmid)

Construction: pACYC184, pKK232-8, kanR

Marker(s):ampR,cmlR,kanR,rbR

Construct size (kb): 8.0

Features: marker(s): ampR
marker(s): cmlR
marker(s): kanR (neoR, rbR)
other: Haemophilus-specific uptake sequence
replicon: p15A
replicon: pMB1
MCS: EcoRI...HindIII
stop codons: 3 reading frames
transcription terminator: T1
transcription terminator: T1 T2

contains sequence

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.

J.M. Musser

Temperature: 37.0°C

Protocol: 5% CO2