Basic Vector Information
- Vector Name:
- pMQ125
- Antibiotic Resistance:
- Gentamicin
- Length:
- 7785 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Source/Author:
- Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
- Promoter:
- araBAD
pMQ125 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 ℃.
pMQ125 vector Sequence
LOCUS 40924_32000 7785 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMQ125, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7785)
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 7785)
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JOURNAL Unpublished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 7785)
AUTHORS Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, O'Toole GA.
TITLE Direct Submission
JOURNAL Submitted (04-MAR-2008) Ophthalmology, University of Pittsburgh, 203
Lothrop St., Pittsburgh, PA 15213, USA
REFERENCE 4 (bases 1 to 7785)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 7785)
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"Unpublished"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
(04-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..7785
/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 3101..3117
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 3127..3145
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature complement(3154..3261)
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(3274..3292)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter complement(3335..3619)
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
CDS 3646..4521
/codon_start=1
/label=araC
/note="L-arabinose regulatory protein"
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
rep_origin 4880..5424
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
CDS complement(5887..6417)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(6606..6634)
/label=Pc promoter
/note="class 1 integron promoter"
promoter complement(6652..6760)
/label=AmpR promoter
misc_feature 6797..7300
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 7565..7785
/label=URA3 promoter
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