pAM34 (ATCC® 77185)

Applications: integrating vectorvector permitting positive selection for inserts  /  Depositors: J Bouche

Permits and Restrictions

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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 (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) - MCS - BamHI - transcription terminator - bla - lacIq.
Media 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

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Shipping Information Distributed: freeze-dried
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