Basic Vector Information
- Vector Name:
- pMELalpha2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7304 bp
- Type:
- UAS-less reporter vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Melcher K, Sharma B, Ding WV, Nolden M.
- Promoter:
- URA3
pMELalpha2 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 ℃.
pMELalpha2 vector Sequence
LOCUS 40924_30590 7304 bp DNA circular SYN 18-DEC-2018 DEFINITION UAS-less reporter vector pMELalpha2, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7304) AUTHORS Melcher K, Sharma B, Ding WV, Nolden M. TITLE Zero background yeast reporter plasmids JOURNAL Gene 247 (1-2), 53-61 (2000) PUBMED 10773444 REFERENCE 2 (bases 1 to 7304) AUTHORS Melcher K, Sharma B, Ding WV, Nolden M. TITLE Direct Submission JOURNAL Submitted (11-SEP-2003) Institute for Microbiology, Frankfurt University, Marie Curie Strasse 9, Frankfurt 60439, Germany REFERENCE 3 (bases 1 to 7304) TITLE Direct Submission REFERENCE 4 (bases 1 to 7304) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene 247 (1-2), 53-61 (2000)" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (11-SEP-2003) Institute for Microbiology, Frankfurt University, Marie Curie Strasse 9, Frankfurt 60439, Germany" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..7304 /mol_type="other DNA" /organism="synthetic DNA construct" regulatory 1..658 /label=UAS deleted MEL1 promoter /note="UAS deleted MEL1 promoter" /regulatory_class="promoter" CDS 659..2071 /gene="MEL1" /label=MEL1 /note="Alpha-galactosidase 1 from Saccharomyces cerevisiae. Accession#: P04824" promoter complement(2471..2489) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2510..2526) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(2534..2550) /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)." promoter complement(2558..2588) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(2603..2624) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(2912..3500) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3674..4531) /label=AmpR /note="beta-lactamase" promoter complement(4532..4636) /label=AmpR promoter misc_feature 4673..5176 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" promoter 5435..5655 /label=URA3 promoter CDS 5656..6456 /label=URA3 /note="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" rep_origin complement(6588..7043) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 7188..7204 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 7211..7229 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 7262..7278 /label=SK primer /note="common sequencing primer, one of multiple similar variants"
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