Basic Vector Information
- Vector Name:
- pMQ236
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5997 bp
- Type:
- Allelic replacement vector
- Replication origin:
- R6K γ ori
- Host:
- Yeast
- Source/Author:
- Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
- Promoter:
- URA3
pMQ236 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 ℃.
pMQ236 vector Sequence
LOCUS 40924_32050 5997 bp DNA circular SYN 18-DEC-2018
DEFINITION Allelic replacement vector pMQ236, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5997)
AUTHORS Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
TITLE New yeast recombineering tools for bacteria
JOURNAL Plasmid 62 (2), 88-97 (2009)
PUBMED 19477196
REFERENCE 2 (bases 1 to 5997)
AUTHORS Shanks RMQ., Kadouri DE, MacEachran DP, O'Toole GA.
TITLE Direct Submission
JOURNAL Submitted (14-MAY-2009) Ophthalmology, University of Pittsburgh, 203
Lothrop St, Pittsburgh, PA 15213, USA
REFERENCE 3 (bases 1 to 5997)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5997)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
date: "2009"; volume: "62"; issue: "2"; pages: "88-97"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(14-MAY-2009) Ophthalmology, University of Pittsburgh, 203 Lothrop
St, Pittsburgh, PA 15213, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5997
/mol_type="other DNA"
/organism="synthetic DNA construct"
oriT complement(352..461)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
terminator complement(882..909)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(1001..1087)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 1437..1453
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(1457..1513)
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(1523..1539)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1547..1563)
/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(1571..1601)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1616..1637)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter complement(1688..1706)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 1914..2302
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS 2653..3444
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
misc_feature 3463..3492
/label=I-SceI meganuclease site
/note="I-SceI meganuclease site"
CDS 3761..4132
/gene="rpsL"
/label=rpsL
/note="Small ribosomal subunit protein uS12 from Salmonella
paratyphi A (strain ATCC 9150 / SARB42). Accession#:
Q5PIW1"
CDS complement(4398..5198)
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
promoter complement(5199..5419)
/label=URA3 promoter
misc_feature 5447..5950
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
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