Basic Vector Information
- Vector Name:
- pMQ117
- Antibiotic Resistance:
- Gentamicin
- Length:
- 5900 bp
- Type:
- Cloning vector
- Replication origin:
- pSC101 ori
- Host:
- Yeast
- Source/Author:
- Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
- Promoter:
- URA3
pMQ117 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 ℃.
pMQ117 vector Sequence
LOCUS 40924_31970 5900 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMQ117, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5900)
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 5900)
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JOURNAL Unpublished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 5900)
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 5900)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 5900)
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"Unpublished"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..5900
/mol_type="other DNA"
/organism="synthetic DNA construct"
oriT complement(399..508)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
terminator complement(929..956)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(1048..1134)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 1484..1500
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(1504..1560)
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(1570..1586)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1594..1610)
/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(1618..1648)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1663..1684)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin 1871..2093
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS 2141..3088
/codon_start=1
/label=Rep101(Ts)
/note="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
/translation="MSELVVFKANELATSRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
CDS complement(3379..3909)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(4098..4126)
/label=Pc promoter
/note="class 1 integron promoter"
promoter complement(4144..4248)
/label=AmpR promoter
CDS complement(4348..5148)
/codon_start=1
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(5149..5369)
/label=URA3 promoter
misc_feature 5397..5900
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
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