pMD-AOX-Laccase vector (V014320)

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

pMD-AOX-Laccase7433 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200AOX1 promoteralpha-factor secretion signalLaccase-2 from Cerrena unicolor. Accession#: P85149Myc6xHisAOX1 terminatorKanRCYC1 terminatorM13 fwdAmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 rev

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

Copy Sequence

Download GeneBank File(.gb)

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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