Basic Vector Information
- Vector Name:
- pMD-AOX-Laccase
- Antibiotic Resistance:
- Kanamycin
- Length:
- 7433 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Yeast
- Selection Marker:
- Neo/G418
- Promoter:
- AOX1
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pMD-AOX-Laccase 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 ℃.
pMD-AOX-Laccase vector Sequence
LOCUS 62056_16630 7433 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7433) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..7433 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 2..937 /label=AOX1 promoter /note="inducible promoter, regulated by methanol" CDS 949..1215 /label=alpha-factor secretion signal /note="N-terminal secretion signal from S. cerevisiae alpha-factor" CDS 1228..1257 /note="Laccase-2 from Cerrena unicolor. Accession#: P85149" CDS 2734..2763 /label=Myc /note="Myc (human c-Myc proto-oncogene) epitope tag" CDS 2779..2796 /label=6xHis /note="6xHis affinity tag" terminator 2876..3122 /label=AOX1 terminator /note="transcription terminator for AOX1" CDS 3265..4077 /label=KanR /note="aminoglycoside phosphotransferase" terminator 4153..4400 /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(4446..4462) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 4936..5040 /label=AmpR promoter CDS 5041..5898 /label=AmpR /note="beta-lactamase" rep_origin 6072..6660 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 6948..6969 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 6984..7014 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7022..7038 /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)." primer_bind 7046..7062 /label=M13 rev /note="common sequencing primer, one of multiple similar variants"
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