pJIR418 [JIR1961] (ATCC® 77387)

Applications: expression vectorshuttle vectorvector permitting construction of fusion proteinsvector permitting visual detection of recombinants  /  Depositors: JI Rood

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Designations pJIR418 [JIR1961]
Depositors JI Rood
Biosafety Level 1
Vector Information
Size (kb): 7.3579998016357420
Vector: pJIR418 (plasmid)
Promoters: Promoter lac
Construction: pUC18, pIP404 replicon, pJIR229 ermBP, pJIR62 catP
Marker(s):cmlR,eryR
Construct size (kb): 7.357999801635742
Features: insert detection: lacZ'
marker(s): cmlR, eryR
promoter: lac
replicon: pMB1, pIP404
Applications
expression vector
shuttle vector
vector permitting construction of fusion proteins
vector permitting visual detection of recombinants
Comments
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).
Media ATCC Medium 1065 (see below) plus 30.0 mcg/ml chloramphenicol ATCC Medium 1065: Tryptone (Difco 0123), 10.0 g Yeast Extract (Difco 0127), 5.0 g NaCl, 10.0 g Distilled water, 1.0 L
Growth Conditions
Temperature: 37.0°C
References

Sloan J, et al. Construction of a sequenced Clostridium perfringens-Escherichia coli shuttle plasmid. Plasmid 27: 207-219, 1992. PubMed: 1513878

Shipped frozen
Shipping Information  Freeze dried  Escherichia coli containing the plasmid.
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