Basic Vector Information
- Vector Name:
- pMQ52
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7277 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, O'toole GA.
- Promoter:
- TEF
pMQ52 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 ℃.
pMQ52 vector Sequence
LOCUS 40924_32110 7277 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle vector pMQ52, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7277)
AUTHORS Shanks RM, Caiazza NC, Hinsa SM, Toutain CM, O'toole GA.
TITLE Saccharomyces cerevisiae-Based Molecular Tool Kit for Manipulation
of Genes from Gram-Negative Bacteria
JOURNAL Appl. Environ. Microbiol. 72 (7), 5027-5036 (2006)
PUBMED 16820502
REFERENCE 2 (bases 1 to 7277)
AUTHORS Shanks RMQ., Caiazza NC, Hinsa SM, Toutain CM, O'Toole GA.
TITLE Direct Submission
JOURNAL Submitted (16-SEP-2005) Microbiology and Immunology, Dartmouth
Medical School, Vail Building Room 505, Hanover, NH 03755, USA
REFERENCE 3 (bases 1 to 7277)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7277)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2006"; volume: "72"; issue: "7"; pages:
"5027-5036"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-SEP-2005) Microbiology and Immunology, Dartmouth Medical School,
Vail Building Room 505, Hanover, NH 03755, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7277
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1..801
/codon_start=1
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
misc_feature 964..997
/label=loxP
/note="loxP"
protein_bind 964..997
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
gene complement(1060..2416)
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
protein_bind complement(2471..2504)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin complement(2555..2906)
/direction=LEFT
/label=pRO1600 oriV
/note="broad-host-range origin of replication from
Pseudomonas aeruginosa plasmid pRO1600; requires the
pRO1600 Rep protein for replication (West et al., 1994)"
CDS 2920..3750
/codon_start=1
/label=pRO1600 Rep
/note="replication protein for the broad-host-range plasmid
pRO1600 from Pseudomonas aeruginosa"
/translation="VASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
primer_bind 3914..3930
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 3940..3958
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature complement(3967..4074)
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(4087..4105)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4126..4142)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4150..4166)
/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(4174..4204)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4219..4240)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4528..5116)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5290..6147)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6148..6252)
/label=AmpR promoter
misc_feature 6289..6792
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 7057..7277
/label=URA3 promoter
This page is informational only.