Basic Vector Information
- Vector Name:
- pEMBLyex-kanMX4
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10195 bp
- Type:
- Yeast expression vector
- Replication origin:
- ori
- Host:
- Yeast
- Source/Author:
- De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R.
- Promoter:
- URA3
pEMBLyex-kanMX4 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 ℃.
pEMBLyex-kanMX4 vector Sequence
LOCUS 40924_17374 10195 bp DNA circular SYN 17-DEC-2018
DEFINITION Yeast expression vector pEMBLyex-kanMX4, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10195)
AUTHORS De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman
Fonseca M, Vanhoucke M, Beyaert R.
TITLE BCCM/LMBP Plasmid collection
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 10195)
AUTHORS De Schamphelaire W.
TITLE Direct Submission
JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent,
Technologiepark 927, 9052, BELGIUM
REFERENCE 3 (bases 1 to 10195)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10195)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
9052, BELGIUM"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10195
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 142..282
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(696..1209)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 1697..1713
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(1980..3071)
/codon_start=1
/label=LEU2
/note="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
rep_origin complement(3429..4775)
/direction=LEFT
/label=2u ori
/note="yeast 2u plasmid origin of replication"
misc_feature complement(4778..4834)
/label=MCS
/note="pUC18/19 multiple cloning site"
protein_bind 5161..5278
/label=UAS
/note="upstream activating sequence mediating
Gal4-dependent induction"
CDS complement(5538..6338)
/codon_start=1
/label=URA3
/note="orotidine-5'-phosphate decarboxylase, required for
uracil biosynthesis"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(6339..6555)
/label=URA3 promoter
primer_bind complement(6575..6591)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6599..6615
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(6623..6653)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6668..6689)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6977..7565)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7738..8595)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8596..8700)
/label=AmpR promoter
gene complement(8734..10090)
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
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