Last data update: 24 January 2024 16:39 CET
Plasmid name: pTACLSPA4 (LMBP 1949)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p1949.gb
(View with Genome Compiler) p1949.txt p1949.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Escherichia coli λ receptor protein cDNA (lamB); with spaA inserts Staphylococcus aureus protein A cDNA (spa); A domain (spaA) Escherichia coli lac repressor gene (lacI) |
| Promoter: | Escherichia coli hybrid tryptophan/lacUV5 promoter (tac) Escherichia coli lac repressor promoter; mutant (lacI(q)) |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) Ribosome binding site (RBS) of the Escherichia coli lambda receptor protein gene (lamB) |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pAJC264; pMcSPA2 |
| Further information: | The plasmid was constructed by inserting four times the small BamHI fragment of pMcSPA2 into the unique BamHI site of pAJC264. As a result, the spaA domain was inserted four times into the lamB coding sequence. The construct is used for studies of exposition of SPA domains on the surface of E. coli. The lacI gene is provided with the lacI(q) promoter, which results in lacI overexpression. Expression of the lamB-spaA fusion gene is rapidly induced by addition of IPTG, which inactivates the lacI repressor. Since the nucleotide sequence at the 5' and at the 3' end of the lamB insert could not be completely traced back, there is uncertainty as to the cutting frequency of the restriction enzymes, except for the sites that were analysed in the authenticity test. The nucleotide sequence of the lamB cDNA corresponds with the EMBL Nucleotide Sequence Database accession number M26131.1. The nucleotide sequence of the spa cDNA corresponds with the EMBL Nucleotide Sequence Database accession number CP000730.1. Other name of the plasmid is pAJC264SPA-A4. |
| EMBL Accession number: | M26131.1, view at EMBL, GenBank, DDBJ CP000730.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 30/09/2008 |
| Sequence detail: | Nucleotide sequence at the borders of the spaA inserts in the lamB coding sequence:
----- lamB ----> ------------------ spaA ------------------>
175 178 169 170 220 224
5' ... GGT.GGT.TCT.TCC.CGG.GAT.CCC.AAT.TTC.AAC.AAA.GAA ... CAA.GCA.CCG.AAA.GCG.GAT.CCC
Gly Gly Ser Ser Arg Asp Pro Asn Phe Asn Lys Glu Gln Ala Pro Lys Ala Asp Pro
AvaI BamHI BamHI
------------------ spaA ------------------> ---------------------- spaA
169 170 220 224 169 170 220
AAT.TTC.AAC.AAA.GAA ... CAA.GCA.CCG.AAA.GCG.GAT.CCC.AAT.TTC.AAC.AAA.GAA ... CAA
Asn Phe Asn Lys Glu Gln Ala Pro Lys Ala Asp Pro Asn Phe Asn Lys Glu Gln
BamHI
--------------> ------------------ spaA ------------------>
224 169 170 220 224
GCA.CCG.AAA.GCG.GAT.CCC.AAT.TTC.AAC.AAA.GAA ... CAA.GCA.CCG.AAA.GCG.GAT.CCC.GCC
Ala Pro Lys Ala Asp Pro Asn Phe Asn Lys Glu Gln Ala Pro Lys Ala Asp Pro Ala
BamHI BamHI
----- lamB ---->
179 180
TCT.TTC.GCC.AGC ... 3'
Ser Phe Ala Ser
Punctuation indicates reading frame. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: AvaI, BamHI, HindII, HindIII and XmnI. The plasmid seems to be approx. 1200 bp larger as compared to the compiled nucleotide sequence, the extra nucleotides being located in the vector part of the plasmid. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr E. Remaut(1) (2). It was constructed by L. Steidler(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., Mol. Gen. Genet. 236 (1993), 187-192 [PMID: 8437563] |
| Related plasmid reference: | Boulain et al., Mol. Gen. Genet. 205 (1986), 339-348 [PMID: 3027509] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pTACLSPA4 (LMBP 1949) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr E. Remaut and was published in Steidler et al., 1993. |
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.