Basic Vector Information
- Vector Name:
- pMK4
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5585 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Zeigler DR.
pMK4 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 ℃.
pMK4 vector Sequence
LOCUS 40924_31005 5585 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pMK4, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5585)
AUTHORS Zeigler DR.
TITLE pMK3 and pMK4 revisited: nucleotide sequences of first generation
Bacillus subtilis - E. coli shuttle vectors
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5585)
AUTHORS Zeigler DR.
TITLE Direct Submission
JOURNAL Submitted (06-MAR-2008) Bacillus Genetic Stock Center, The Ohio
State University, 484 W 12th Ave, Columbus, OH 43210, USA
REFERENCE 3 (bases 1 to 5585)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5585)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-MAR-2008) Bacillus Genetic Stock Center, The Ohio State
University, 484 W 12th Ave, Columbus, OH 43210, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5585
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 106..127
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 142..172
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 180..196
/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 204..220
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(269..285)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 759..863
/label=AmpR promoter
CDS 864..1721
/label=AmpR
/note="beta-lactamase"
CDS complement(2460..2474)
/label=enterokinase site
/note="enterokinase recognition and cleavage site"
CDS complement(2572..3507)
/codon_start=1
/gene="repC"
/product="replication initiation protein"
/label=repC
/note="RepC; initiation of plasmid replication; derived
from Staphylococcus aureus plasmid pC194"
/protein_id="ACB41735.1"
/translation="MCYNMEKYTEKKQRNQVFQKFIKRHIGENQMDLVEDCNTFLSFVA
DKTLEKQKLYKANSCKNRFCPVCAWRKARKDALGLSLMMQYIKQQEKKEFIFLTLTTPN
VMSDELENEIKRYNNSFRKLIKRKKVGSVIKGYVRKLEITYNKKRDDYNPHFHVLIAVN
KSYFTDKRYYISQQEWLDLWRDVTGISEITQVQVQKIRQNNNKELYEMAKYSGKDSDYL
INQKVFDAFYKSLKGKQVLVYSGLFKEAKKKLKNGDLDYLKEIDPTEYIYQIFYIWKQK
EYLASELYDLTEQEKREINHKMIDEIEEEQ"
gene complement(2572..3507)
/gene="repC"
/label=repC
CDS 3965..4612
/gene="cat"
/label=cat
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
rep_origin 4815..5403
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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