Basic Vector Information
- Vector Name:
- pMQ123
- Antibiotic Resistance:
- Gentamicin
- Length:
- 9582 bp
- Type:
- Cloning vector
- Replication origin:
- oriV
- Host:
- Yeast
- Source/Author:
- Shanks RM, MacEachran DP, Powers SZ, Kadouri DE, O'Toole GA.
- Promoter:
- URA3
pMQ123 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 ℃.
pMQ123 vector Sequence
LOCUS 40924_31985 9582 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMQ123, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9582)
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JOURNAL Unpublished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 2 (bases 1 to 9582)
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 3 (bases 1 to 9582)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9582)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(04-MAR-2008) Ophthalmology, University of Pittsburgh, 203 Lothrop
St., Pittsburgh, PA 15213, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9582
/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"
rep_origin 1198..1728
/label=oriV
/note="incP origin of replication"
CDS 2120..3265
/codon_start=1
/label=trfA
/note="trans-acting replication protein that binds to and
activates oriV"
/translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
terminator complement(3525..3611)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(3841..4554)
/codon_start=1
/label=yeGFP
/note="yeast-enhanced green fluorescent protein"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
protein_bind complement(4653..4669)
/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(4677..4705)
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
promoter 4935..5012
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 5136..5951
/codon_start=1
/gene="lacI"
/product="truncated LacI"
/label=lacI
/protein_id="ACB45224.1"
/translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTTWTACCNSLRARR"
gene 5136..5951
/gene="lacI"
/label=lacI
rep_origin complement(6132..6720)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6932..7462)
/codon_start=1
/label=GmR
/note="gentamycin acetyltransferase"
/translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
EEVMHFDIDPSTAT"
promoter complement(7651..7679)
/label=Pc promoter
/note="class 1 integron promoter"
rep_origin complement(7756..9098)
/direction=LEFT
/label=2u ori
/note="yeast 2u plasmid origin of replication"
promoter 9362..9582
/label=URA3 promoter
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