Basic Vector Information
- Vector Name:
- pMAK705
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 5593 bp
- Type:
- Cloning vector
- Replication origin:
- pSC101 ori
- Source/Author:
- Hamilton CM, Aldea M, Washburn BK, Babitzke P, Kushner SR.
pMAK705 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 ℃.
pMAK705 vector Sequence
LOCUS 40924_29686 5593 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMAK705, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5593)
AUTHORS Hamilton CM, Aldea M, Washburn BK, Babitzke P, Kushner SR.
TITLE New method for generating deletions and gene replacements in
Escherichia coli
JOURNAL J. Bacteriol. 171 (9), 4617-4622 (1989)
PUBMED 2548993
REFERENCE 2 (bases 1 to 5593)
AUTHORS Maples VF, Kushner SR.
TITLE Direct Submission
JOURNAL Submitted (06-JUN-2002) Genetics, University of Georgia, Life
Sciences Bldg., Athens, GA 30602, USA
REFERENCE 3 (bases 1 to 5593)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5593)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J.
Bacteriol."; date: "1989"; volume: "171"; issue: "9"; pages:
"4617-4622"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-JUN-2002) Genetics, University of Georgia, Life Sciences Bldg.,
Athens, GA 30602, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5593
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 487..709
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS 757..1704
/label=Rep101(Ts)
/note="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
promoter complement(3191..3295)
/label=AmpR promoter
promoter 3498..3600
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 3601..4257
/label=CmR
/note="chloramphenicol acetyltransferase"
primer_bind 5205..5221
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 5222..5278
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(5291..5307)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5315..5331)
/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)."
promoter complement(5339..5369)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5384..5405)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 5506..5593
/note="derived from vector pBR325 deposited in GenBank
Accession Number L08855"
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