Basic Vector Information
- Vector Name:
- pMQ156
- Antibiotic Resistance:
- Gentamicin
- Length:
- 6032 bp
- Type:
- Cloning vector
- Replication origin:
- R6K γ ori
- Host:
- Yeast
- Source/Author:
- Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
- Promoter:
- URA3
pMQ156 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 ℃.
pMQ156 vector Sequence
LOCUS 40924_32020 6032 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMQ156, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6032)
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 6032)
AUTHORS Shanks RMQ., Kadouri DE, O'Toole GA.
TITLE Recombineering vectors for a wide range of Gram-negative and
-positive bacteria
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 6032)
AUTHORS Shanks RMQ., Kadouri DE, O'Toole GA.
TITLE Direct Submission
JOURNAL Submitted (13-OCT-2008) Ophthalmololgy, University of Pittsburgh,
203 Lothrop St, Pittsburgh, PA 15213, USA
REFERENCE 4 (bases 1 to 6032)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6032)
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"Unpublished"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
(13-OCT-2008) Ophthalmololgy, University of Pittsburgh, 203 Lothrop
St, Pittsburgh, PA 15213, USA"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6032
/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"
oriT complement(893..1001)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
protein_bind complement(1198..1231)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin complement(1282..1633)
/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 1647..2477
/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"
terminator complement(2679..2765)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 3115..3131
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(3135..3191)
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(3201..3217)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3225..3241)
/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(3249..3279)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3294..3315)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter complement(3366..3384)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 3593..3981
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(4134..4664)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(4853..4881)
/label=Pc promoter
/note="class 1 integron promoter"
promoter complement(4899..5007)
/label=AmpR promoter
misc_feature 5044..5547
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 5812..6032
/label=URA3 promoter
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