Restriction digests of the clone give the following sizes (kb): EcoRI--9.3; EcoRV--5.8, 3.6; SspI--3.9, 1.8, 1.6, 1.2, 0.9; EcoRI/MluI--8.4, 0.9.
Contains sequences encoding lacIq, a tandem tac lacUV5 promoter region, and lacZalpha to allow direct screening by inactivation of alpha-complementation. Shows a low level of expression without induction.
Sequencing demonstrated that there are 3 stop codons in frame with the truncated bla gene, terminating translation but permitting continued transcription and expression of the cat gene.
Media
ATCC® Medium 1675: LB medium (ATCC medium 1065) with 10 mcg/ml Chloramphenicol
Growth Conditions
Temperature: 37°C
References
Morales VM, et al. A series of wide-host-range low-copy-number vectors that allow direct screening for recombinants. Gene 97: 39-47, 1991. PubMed: 1847347