Restriction digests of the clone give the following sizes (kb): EcoRI--7.4; HindIII--7.4; SmaI--5.0, 2.4.
The vector contains the following restriction sites (position in bp): EcoRI--1; HindIII--57; PvuI--175; FspI--195; NarI--216; Tth111I--397; SpeI--3235; NheI--3549; NaeI--3653; HpaI--4206; EcoRV--4723; SmaI--4984; AvaII--5059; ScaI--5850.
The genes for chloramphenicol and erythromycin resistance (catP and ermBP, respectively) were cloned from C. perfringens and can be used for selection in both C. perfringens and E. coli.
EcoRV and ScaI restriction sites can be used for cloning and insertional inactivation of catP and ermBP, respectively.
The standard reverse sequencing primer is not useful in this vector due to the duplication of the 181 bp region between pUC18 sites PvuII and EcoRI.
Shuttle expression vector permitting visual detection of recombinants in E. coli using lacZalpha complementation. Does not contain an intact beta-lactamase (bla) gene.
The order of the major features in this plasmid is: 5' lacZ' - EcoRI/MCS/HindIII - 3' lacZ' - pIP404 ori - pIP404 rep - HpaI - catP - SmaI - AvaII - ermBP - pMB1 ori.
Location of the major features in this plasmid: lacZ' (nt 7345-310), MCS (nt 1-57), pIP404 ori (approx. nt 1190-1419), pIP404 rep gene (nt 2702-1482), catP (nt 4830-4207), ermBP (nt 5902-5165), pMB1 ori (approx. nt 6780-7000). |
