Basic Vector Information
- Vector Name:
- pMQ2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8636 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
- Promoter:
- URA3
pMQ2 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 ℃.
pMQ2 vector Sequence
LOCUS 40924_32030 8636 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMQ2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8636)
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 8636)
AUTHORS Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, O'Toole GA.
TITLE In vivo recombination vectors for modification and expression of
genes in Gram-negative and Gram-positive bacteria
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 8636)
AUTHORS Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, O'Toole GA.
TITLE Direct Submission
JOURNAL Submitted (03-MAR-2008) Ophthalmology, University of Pittsburgh, 203
Lothrop St., Pittsburgh, PA 15213, USA
REFERENCE 4 (bases 1 to 8636)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 8636)
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:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(03-MAR-2008) Ophthalmology, University of Pittsburgh, 203 Lothrop
St., Pittsburgh, PA 15213, USA"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8636
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 196..416
/label=URA3 promoter
CDS 417..1217
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
rep_origin complement(1351..1806)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 1951..1967
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 1977..1995
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 2021..2037
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 2274..2431
/label=palA lagging strand origin
/note="palA lagging strand origin"
CDS complement(2573..3220)
/gene="cat"
/label=cat
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
CDS complement(3389..3772)
/codon_start=1
/gene="rep"
/product="replication protein"
/label=rep
/protein_id="ACB45180.1"
/translation="MHLILSNMTFNKFLKYLISLFFLSILFHVITHKNNLVFTNYDNKK
SCFFPFLCMFFTSHLKRYINRYEKATFFALKTSHTNNLRVTSLAGNSYPYYQDKKEKDF
SLRSNPLKKHKRPHFLMWSLFFN"
gene complement(3389..3772)
/gene="rep"
/label=rep
rep_origin 3427..3555
/label=oriV for pC194
/note="oriV for pC194"
CDS 3677..4366
/codon_start=1
/gene="pre"
/product="Pre"
/label=pre
/note="pC194 OrfA"
/protein_id="ACB45181.1"
/translation="MCYNMEKYTEKKQRNQVFQKFIKRHIGENQMDLVEDCNTFLSFVA
DKTLEKQKLYKANSCKNRFCPVCAWRKARKDALGLSLMMQYIKQQEKKEFIFLTLTTPN
VMSDELENEIKRYNNSFRKLIKRKKVGSVIKGYVRKLEITYNKKRDDYNPHFHVLIAVN
KSYFTDKRYYISQQEWLDLWRDVTGISEITQVQVQKIRQNNNKELYEMAKYSGKDSDYL
INKSKSL"
gene 3677..4366
/gene="pre"
/label=pre
misc_feature 4613..4699
/label=palB
/note="palB"
CDS 4713..4727
/label=enterokinase site
/note="enterokinase recognition and cleavage site"
primer_bind complement(4981..4997)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(5034..5052)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5073..5089)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5097..5113)
/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(5121..5151)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5166..5187)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5475..6063)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6237..7094)
/label=AmpR
/note="beta-lactamase"
promoter complement(7095..7199)
/label=AmpR promoter
rep_origin complement(7226..8568)
/direction=LEFT
/label=2u ori
/note="yeast 2u plasmid origin of replication"
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