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LMBP plasmid details

Last data update: 24 November 2020 04:18 CET

Plasmid name: pPAS8 (LMBP 2131)

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Price category: Cat. 1 (cf. price list)
Status: GeneCorner non-core plasmid
Depositor's sequence: ppas8.gb
Sequence
analysis results
Genecorner:

-

Cloned DNA: Mutant 68B of Escherichia coli papA gene (papAm) - A domain of Staphylococcus aureus spa gene fusion
Promoter: Phage λ major leftward promoter (λ PL)
Ribosome
binding site:
Ribosome binding site (RBS) of the phage MS2 polymerase gene; hybrid with papA SD
Terminator: -
Selection marker: Ampicillin (amp)
Replicon: Escherichia coli plasmid pMB1 origin
Host range: Escherichia coli; use strains with a cI function, cIts for PL controlled expression
Parental clone: pPAS6; pMcSPA2
Further information: This plasmid was constructed as follows : the small BamHI fragment of pMcSPA2 (containing the A domain of the S. aureus spa gene) was inserted into the unique BamHI site, created at AA 68 of the papaA gene, of pPAS6.
Other name of the plasmid is pPLc245PAPA68BSPAA.
EMBL Accession number: -
Latest sequence update: 10/06/1993
Sequence detail:
Nucleotide sequence around the gene fusion:



            papA ->|        |<- spa
                   |        |
  64  65  66  67  68 linker  111 112            
 ACT GCC TTT AAA GGG GAT CCC AAT TTC AAC AAA GAA CAA CAA AAT GCT
 Thr Ala Phe Lys Gly Asp Pro Asn Phe Asn Lys Glu Gln Gln Asn Ala 
                   BamHI
 120                                     130
 TTC TAT GAA ATC TTG AAT ATG CCT AAC TTA AAC GAA GAA CAA CGC AAT
 Phe Tyr Glu Ile Leu Asn MET Pro Asn Leu Asn Glu Glu Gln Arg Asn 
                  140                                 150
 GGT TTC ATC CAA AGC TTA AAA GAT GAC CCA AGC CAA AGT GCT AAC CTA
 Gly Phe Ile Gln Ser Leu Lys Asp Asp Pro Ser Gln Ser Ala Asn Leu 
                                                          spa ->|
                                                                |
                                                    A ->|<- B
                                 160                 165|166 167
 TTG TCA GAA GCT AAA AAG TTA AAT GAA TCT CAA GCA CCG AAA GCG GAT
 Leu Ser Glu Ala Lys Lys Leu Asn Glu Ser Gln Ala Pro Lys Ala Asp 
                                                           BamHI
       |<- papA
       |
 linker  71  72  73  74
    CCT GGC GCC AAA AAA 
    Pro Gly Ala Lys Lys

                                                            
 
Nucleotide sequence of the (mutated) A domain of S. aureus
spa gene in pMcSPA2 :


            D ->|<- A
                |        110            
 CAA GCA CCG AAA GCG GAT CCC AAT TTC AAC AAA GAA CAA CAA AAT GCT
 Gln Ala Pro Lys Ala Asp Pro Asn Phe Asn Lys Glu Gln Gln Asn Ala 
                   BamHI

 120                                     130
 TTC TAT GAA ATC TTG AAT ATG CCT AAC TTA AAC GAA GAA CAA CGC AAT
 Phe Tyr Glu Ile Leu Asn MET Pro Asn Leu Asn Glu Glu Gln Arg Asn 

                 140                                 150
 GGT TTC ATC CAA AGC TTA AAA GAT GAC CCA AGC CAA AGT GCT AAC CTA
 Gly Phe Ile Gln Ser Leu Lys Asp Asp Pro Ser Gln Ser Ala Asn Leu 

                                                    A ->|<- B
                                 160                    |
 TTG TCA GAA GCT AAA AAG TTA AAT GAA TCT CAA GCA CCG AAA GCG GAT
 Leu Ser Glu Ala Lys Lys Leu Asn Glu Ser Gln Ala Pro Lys Ala Asp 
                                                           BamHI

         170
 CCC AAA TTC AAC AAA GAA  
 Asn Lys Phe Asn Lys Glu
Authenticity test: The plasmid still needs to be subjected to the authenticity test.
Class: Recombinant plasmid
Type: Plasmid
History of deposit: This plasmid was deposited by Dr L. Steidler(1) (2) and Prof. Dr E. Remaut(1) (2).
(1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium
(2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium
Plasmid reference: Steidler et al., J. Bacteriol. 175 (1993), 7639-7643 [PMID: 7902348]
Restricted distribution: - BCCM MTA
Distributed as: H/P active culture or plasmid DNA
Host for distribution: Escherichia coli K12 MC1061[pcI857]
Host reference: Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493]
Helper plasmid: pcI857
Cultivation medium: LB-Lennox + ampicillin (100 μg/ml) + kanamycin (50 μg/ml)
Cultivation temperature: 28°C
Biosafety level: L1
Cultivation remark: -
Other culture collection numbers: LMBP 02129

Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.

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