Basic Vector Information
- Vector Name:
- pMHE3Tc
- Antibiotic Resistance:
- Tetracycline
- Length:
- 6384 bp
- Type:
- Broad host range expression vector
- Replication origin:
- pBBR1 oriV
- Source/Author:
- Fodor BD, Kovacs AT, Csaki R, Hunyadi-Gulyas E, Klement E, Maroti G, Meszaros LS, Medzihradszky KF, Rakhely G, Kovacs KL.
pMHE3Tc vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pMHE3Tc vector Sequence
LOCUS 40924_30825 6384 bp DNA circular SYN 18-DEC-2018 DEFINITION Broad host range expression vector pMHE3Tc, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6384) AUTHORS Fodor BD, Kovacs AT, Csaki R, Hunyadi-Gulyas E, Klement E, Maroti G, Meszaros LS, Medzihradszky KF, Rakhely G, Kovacs KL. TITLE Modular broad-host-range expression vectors for single-protein and protein complex purification JOURNAL Appl. Environ. Microbiol. 70 (2), 712-721 (2004) PUBMED 14766546 REFERENCE 2 (bases 1 to 6384) AUTHORS Fodor BD. TITLE Direct Submission JOURNAL Submitted (16-MAY-2003) Biophysics, BRC Hungarian Academy of Sciences, Temesvari krt., Szeged, Csongrad 6701, Hungary REFERENCE 3 (bases 1 to 6384) TITLE Direct Submission REFERENCE 4 (bases 1 to 6384) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl. Environ. Microbiol."; date: "2004"; volume: "70"; issue: "2"; pages: "712-721" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (16-MAY-2003) Biophysics, BRC Hungarian Academy of Sciences, Temesvari krt., Szeged, Csongrad 6701, Hungary" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6384 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 198..216 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 217..241 /label=lac operator /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 256..278 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 286..318 /label=T7 tag (gene 10 leader) /note="leader peptide from bacteriophage T7 gene 10" CDS 367..384 /label=6xHis /note="6xHis affinity tag" terminator 451..498 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 535..963 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(1526..2185) /label=pBBR1 Rep /note="replication protein for the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica" rep_origin complement(2186..2955) /direction=LEFT /label=pBBR1 oriV /note="replication origin of the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica; requires the pBBR1 Rep protein for replication" misc_feature 3179..4172 /note="RP4 mob; oriT" regulatory complement(4262..4377) /label=T4 terminator /note="T4 terminator" /regulatory_class="terminator" CDS complement(4686..5873) /label=TcR /note="tetracycline efflux protein" regulatory 6258..6373 /label=T4 terminator /note="T4 terminator" /regulatory_class="terminator"
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