pPLc24 [PL-A] (ATCC® 31697)

Applications: expression vector  /  Depositors: Biogen, Inc., JF Haley, Biogen, Inc.

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Designations pPLc24 [PL-A]
Depositors Biogen, Inc., JF Haley, Biogen, Inc.
Biosafety Level 1
Host
Distribution host: Escherichia coli M5219
Distribution host: Escherichia coli M5219
Vector Information
Size (kb): 3.1429998874664310
Vector: pPLc24 (plasmid)
Promoters: Promoter lambda PL
Construction: pBR322, MS2, lambda
Marker(s):ampR
Construct size (kb): 3.142999887466431
Features: marker(s): ampR
promoter: lambda PL
replicon: pMB1
enhancer: none
Applications
expression vector
Comments
The following unique restriction sites are found on this vector separated by (bp)(approx): BglI- 100- BamHI- 900- EcoRI- 600- XhoI- 300- SmaI- 250- HindIII- 1650.
Restriction digests of the clone give the following sizes (kb): EcoRI--3.1; PvuI/BamHI--1.7, 1.4; BglI--3.1; PstI--3.1; HindIII--3.1.
Plates equally well at 28C and 42C in E. coli K-12 deltaH1 hosts.
Escherichia coli M5219 is Escherichia coli K-12 M72 lac(am) trp(am) rpsL lambda cI857 deltaH1 bio252. deltaH1 removes part of cro and all genes to the right of cro. bio252 removes all genes to the left of cIII. At 42C, N is expressed from the chromosome.
Translation from the MS2 replicase is colinear with transcription from the PL promoter and thus is under PL control.
Shows reduced plating efficiency in E. coli M5219 at 42C under antibiotic selection.
This vector is used for expression of fused proteins with MS2 polymerase.
ATG is in phase with GAT from the BamHI site, AAG from the HindIII site, TTA from the MstII site, CGC from the NruI site and GCT from the EspI site.
The orientation of the PL promoter is clockwise with respect to the plasmid ori.
This vector was constructed from pPLc28 (ATCC 31696) by inserting a 431 bp EcoRI/BamHI fragment coding for the ribosome binding site and the first 98 amino acids of the MS2 replicase (from pMS2-7) into pPLc28.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 28.0°C
References

Fiers WC, Remaut ER. Vectors and methods for making such vectors and for expressing cloned genes. US Patent 4,874,702 dated Oct 17 1989

Remaut E, et al. Plasmid vectors for high-efficiency expression controlled by the PL promoter of coliphage lambda. Gene 15: 81-93, 1981. PubMed: 6271633

Shipped freeze-dried
Shipping Information Distributed: freeze-dried
Product Sheet
Product Sheet
Product Sheet