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| 产品名称： | pAM34 |
|---|---|
| 商品货号： | TS133870 |
| Designations： | pAM34 |
| Depositors： | J Bouche |
| Biosafety Level： | 1 |
| Host： | Distribution host: Escherichia coli JS219 |
| Vector Information： | Size (kb): 6.0000000000000000 DESCRIPTION OF VECTOR: Intact vector size: 6.000 Type of vector: plasmid Cloning sites: BamHI SalI PstI SphI XmaIII HindIII EcoRI SacI KpnI Polylinker sites: BamHI SalI PstI SphI HindIII EcoRI SacI KpnI BamHI Construction: pBR322, R100.1, pCK1 Host range: Escherichia coli Features (with orientation and position when available): insert detection: T4gene32 ter marker(s): ampR, spcR replicon: pMB1 repressor gene: lacIq Vector: pAM34 (plasmid) Construction: pBR322, R100.1, pCK1 Marker(s):ampR,spcR Construct size (kb): 6.0 Features: insert detection: T4gene32 ter marker(s): ampR, spcR replicon: pMB1 repressor gene: lacIq |
| Applications： | integrating vector vector permitting positive selection for inserts |
| Comments： | Restriction digests of the clone give the following sizes (kb): EcoRI--6.0; BamHI--4.3, 1.8; HindIII--6.0; PstI--6.0; KpnI--3.4, 2.7. Can be selected with 100 ug/ml ampicillin and/or 75 ug/ml spectinomycin. Permits positive selection because deletion of the aadA fragment is not viable. E. coli JS219 is MC1061 malPp delta534::lacIq. Integration vector permitting positive selection for inserts. Transcription of the replication primer RNA is regulated by the lacZpo promoter/operator. In the presence of lacIq, replication is entirely dependent on the presence of inducer. The repressible replication system permits temporary maintenance of the plasmid, construction of strains with stable integrants of vector derivatives, and recovery of sequences adjacent to cloned fragments. pAM35 (ATCC 77186) differs from pAM34 (TS133870) by deletion of the DraII fragment in pAM34 carrying lacIq. The order of the major features in this plasmid is: lacZpo - ori - rom - transcription terminator - BamHI - MCS - XmaIII - aadA (spcR) - MCS - BamHI - transcription terminator - bla - lacIq. |
| Media： | ATCC® Medium 1122: LB with IPTG medium (ATCC medium 1065) with 10 ml/L filter-sterilized 100 mM IPTG (isopropylthio-beta-galactoside) |
| Growth Conditions： | Temperature: 37.0°C |
| References： | Gil D, Bouche JP. ColE1-type vectors with fully repressible replication. Gene 105: 17-22, 1991. PubMed: 1937005 |
| Shipped： | freeze-dried |
| Shipping Information： | Distributed: freeze-dried |