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| 产品名称： | pAM35 |
|---|---|
| 商品货号： | TS145266 |
| Designations： | pAM35 |
| Depositors： | J Bouche |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 4.8000001907348630 Vector: pAM35 (plasmid) Construction: pBR322, R100.1 Marker(s):ampR,spcR Construct size (kb): 4.800000190734863 Features: insert detection: T4gene32 ter marker(s): ampR, spcR replicon: pMB1 repressor gene: lacIq terminator: T4 gene 32 |
| Applications： | vector permitting positive selection for inserts |
| Comments： | Restriction digests of the clone give the following sizes (kb): EcoRI--4.8; BamHI--3.0, 1.8; KpnI--3.4, 1.4. 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. It can be propagated in a lacI+ strain such as C600 in the absence of IPTG, but replication is inefficient. 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 (TS145266) differs from pAM34 (ATCC 77185) 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) - HindIII - MCS - BamHI - transcription terminator - bla. |
| 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 |