Basic Vector Information
- Vector Name:
- pMQ299
- Length:
- 6913 bp
- Type:
- Delivery vector
- Replication origin:
- pSC101 ori
- Host:
- Yeast
- Source/Author:
- Kalivoda EJ, Horzempa J, Stella NA, Sadaf A, Kowalski RP, Nau GJ, Shanks RM.
- Promoter:
- URA3
pMQ299 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 ℃.
pMQ299 vector Sequence
LOCUS 40924_32060 6913 bp DNA circular SYN 18-DEC-2018
DEFINITION Delivery vector pMQ299, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6913)
AUTHORS Kalivoda EJ, Horzempa J, Stella NA, Sadaf A, Kowalski RP, Nau GJ,
Shanks RM.
TITLE New vector tools with a hygromycin resistance marker for use with
opportunistic pathogens
JOURNAL Mol. Biotechnol. 48 (1), 7-14 (2011)
PUBMED 20972648
REFERENCE 2 (bases 1 to 6913)
AUTHORS Kalivoda EJ, Horzempa JE, Sadaf AA, Stella NA, Nau GJ, Shanks RM.
TITLE Direct Submission
JOURNAL Submitted (19-DEC-2009) Ophthalmology, University of Pittsburgh, EEI
1015, 203 Lothrop St, Pittsburgh, PA 15213, USA
REFERENCE 3 (bases 1 to 6913)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6913)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
Biotechnol."; date: "2011"; volume: "48"; issue: "1"; pages: "7-14"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(19-DEC-2009) Ophthalmology, University of Pittsburgh, EEI 1015, 203
Lothrop St, Pittsburgh, PA 15213, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6913
/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"
CDS complement(1524..2216)
/gene="SCEI"
/label=SCEI
/note="Intron-encoded endonuclease I-SceI from
Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
Accession#: P03882"
protein_bind complement(2246..2262)
/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(2270..2300)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2315..2336)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin 2523..2745
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS 2793..3740
/label=Rep101(Ts)
/note="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
CDS complement(4031..5026)
/gene="hyg"
/label=hyg
/note="Hygromycin-B 7''-O-kinase from Streptomyces
hygroscopicus. Accession#: P09979"
promoter complement(5157..5261)
/label=AmpR promoter
CDS complement(5361..6161)
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
promoter complement(6162..6382)
/label=URA3 promoter
misc_feature 6410..6913
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
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