pLDR10 [WM2154] (ATCC® 77359)

Organism: bacteriophage lambda  /  Clone Type: Vector  /  Depositors: W Messer

Permits and Restrictions

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Designations pLDR10 [WM2154]
Species bacteriophage lambda
Depositors W Messer
Applications
contains sequence attachment site
integrating vector
Vector
Size (kb): 3.8
Vector: pLDR10 (plasmid)
Construction: pT7-7, lambda attP, pACYC184 cat
Marker(s): ampR,cmlR
Construct size (kb): 3.8
Features: marker(s): ampR, cmlR
replicon: pMB1
Insert
DNA: genomic
Insert lengths(kb): 0.493
Gene product: attachment site [attP]
Target Gene: attachment site
Insert Size (kb) 0.493
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Related Products
Component of: ATCC 77371
Biosafety Level 1
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI/BglII--1.8, 1.6, 0.5; BamHI--3.9; XbaI--3.9; NotI--2.0, 1.9.
Vector containing the attP sequence for integrating DNA into the lambda attachment site attB. Requires the lambda integrase (int) such as that provided by pLDR8 (ATCC 77357).
When a recombinant construct is digested with NotI, the origin is separated from the insert. Each fragment has a distinct selectable marker. After religation to form origin-free circles, the products are transformed into bacteria with pLDR8.
Expression of int in pLDR8 is regulated by lambda PR, by the temperature sensitive cI857 repressor. Replication of pLDR8 is also temperature sensitive.
Therefore, at 30C replication of the plasmid is normal and int is not expressed. At 42C, int is expressed but pLDR8 is no longer replicated, resulting in loss of int after a few cell generations.
The order of the major features in this plasmid is: NotI - ampR - ClaI/MCS/EcoRI - attP - NotI - pMB1 ori - cmlR.
References

Diederich L, et al. New cloning vectors for integration in the lambda attachment site attB of the Escherichia coli chromosome. Plasmid 28: 14-24, 1992. PubMed: 1387714

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