Basic Vector Information
- Vector Name:
- pMK2016
- Antibiotic Resistance:
- Streptomycin
- Length:
- 6981 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- House BL, Mortimer MW, Kahn ML.
- Promoter:
- lac UV5
pMK2016 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 ℃.
pMK2016 vector Sequence
LOCUS 40924_30955 6981 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMK2016, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6981)
AUTHORS House BL, Mortimer MW, Kahn ML.
TITLE New recombination methods for Sinorhizobium meliloti genetics
JOURNAL Appl. Environ. Microbiol. 70 (5), 2806-2815 (2004)
PUBMED 15128536
REFERENCE 2 (bases 1 to 6981)
AUTHORS House BL, Mortimer MW, Kahn ML.
TITLE Direct Submission
JOURNAL Submitted (29-SEP-2003) School of Molecular Biosciences, Washington
State University, Abelson Hall Room 406A, Pullman, WA 99164, USA
REFERENCE 3 (bases 1 to 6981)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6981)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2004"; volume: "70"; issue: "5"; pages:
"2806-2815"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(29-SEP-2003) School of Molecular Biosciences, Washington State
University, Abelson Hall Room 406A, Pullman, WA 99164, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6981
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 605..1195
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1615..1803)
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
CDS 1843..2187
/gene="mbeC"
/label=mbeC
/note="Mobilization protein MbeC from Escherichia coli.
Accession#: P13657"
primer_bind 3507..3523
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(3526..3582)
/label=MCS
/note="pUC18/19 multiple cloning site"
protein_bind complement(3599..3646)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
protein_bind 3687..3806
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 3836..3866
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 3920..4576
/label=CmR
/note="chloramphenicol acetyltransferase"
CDS 4921..5223
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
protein_bind complement(5267..5391)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
protein_bind 5432..5479
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
misc_feature 5496..5552
/label=MCS
/note="pUC18/19 multiple cloning site"
CDS complement(5803..6591)
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
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