Basic Vector Information
- Vector Name:
- pMDS20
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9983 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Holmes M, Pfeifer F, Dyall-Smith M.
- Promoter:
- T3
pMDS20 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 ℃.
pMDS20 vector Sequence
LOCUS 40924_30445 9983 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pMDS20, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9983)
AUTHORS Holmes M, Pfeifer F, Dyall-Smith M.
TITLE Improved shuttle vectors for Haloferax volcanii including a
dual-resistance plasmid
JOURNAL Gene 146 (1), 117-121 (1994)
PUBMED 8063095
REFERENCE 2 (bases 1 to 9983)
AUTHORS Dyall-Smith ML.
TITLE Direct Submission
JOURNAL Submitted (16-MAR-1994) Microbiology, University of Melbourne,
Grattan Street, Parkville, Victoria, Australia
REFERENCE 3 (bases 1 to 9983)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9983)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene";
date: "1994"; volume: "146"; issue: "1"; pages: "117-121"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-MAR-1994) Microbiology, University of Melbourne, Grattan Street,
Parkville, Victoria, Australia"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9983
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 894..5001
/mol_type="genomic DNA"
/note="Haloferax Aa2.2 replicon region; includes origin of
replication"
/db_xref="taxon:56961"
/organism="Shuttle vector pMDS20"
rep_origin complement(237..692)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 837..853
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 860..878
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 5333..7249
/gene="gyrB"
/label=gyrB
/note="DNA gyrase subunit B from Haloferax lucentense
(strain DSM 14919 / JCM 9276 / NCIMB 13854 / Aa 2.2).
Accession#: P21558"
promoter complement(7723..7741)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(7762..7778)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 7786..7802
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(7810..7840)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(7855..7876)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(8164..8752)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(8926..9783)
/label=AmpR
/note="beta-lactamase"
promoter complement(9784..9888)
/label=AmpR promoter
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