Basic Vector Information
- Vector Name:
- pEKEx2
- Antibiotic Resistance:
- Kanamycin
- Length:
- 8161 bp
- Type:
- Shuttle-expression vector
- Replication origin:
- ori
- Source/Author:
- Krumbach K, Eggeling L, Eikmanns B.
pEKEx2 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 ℃.
pEKEx2 vector Sequence
LOCUS 40924_17299 8161 bp DNA circular SYN 17-DEC-2018
DEFINITION Shuttle-expression vector pEKEx2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8161)
AUTHORS Krumbach K, Eggeling L, Eikmanns B.
TITLE Direct Submission
JOURNAL Submitted (30-MAR-2004) Biotechnology, Forschungszentrum, Leo-Brand,
Juelich, NRW 52425, Germany
REFERENCE 2 (bases 1 to 8161)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8161)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(30-MAR-2004) Biotechnology, Forschungszentrum, Leo-Brand, Juelich,
NRW 52425, Germany"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8161
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(1..57)
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(58..74)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator 287..373
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 465..492
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 511..602
/label=AmpR promoter
primer_bind complement(923..939)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(947..963)
/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(971..1001)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1016..1037)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1325..1913)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 2146..2958
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
CDS 4577..5860
/codon_start=1
/product="replicase"
/label=replicase
/note="orf-1 from Corynebacterium glutamicum pBL1"
/protein_id="AAS99552.1"
/translation="MNTSKEPQVNEGSKVTRARAWRRQNVMYKITNSKALAGCHRWRRD
EAVAVSWSSNGASQFEGLQNSHSRWGSPLAELEVMGERRIELAIATKNHLAAGGALMMF
VGTVRHNRSQSFAQVEAGIKTAYSSMVKTSQWKKERARYGVEHTYSDYEVTDSWANGWH
LHRNMLLFLDRPLSDDELKAFEDSMFSRWSAGVVKAGMDAPLREHGVKLDQVSTWGGDA
AKMATYLAKGMSQELTGSATKTASKGSYTPFQMLDMLADQSDAGEDMDAVLVARWREYE
VGSKNLRSSWSRGAKRALGIDYIDADVRREMEEELYKLAGLEAPERVESTRVAVALVKP
DDWKLIQSDFAVRQYVLDCVDKAKDVAAAQRVANEVLASLGVDSTPCMIVMDDVDLDAV
LPTHGDATKRDLNAAVFAGNEQTILRTH"
promoter complement(6581..6609)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
CDS complement(6724..7752)
/codon_start=1
/label=lacI
/note="lac repressor"
/translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ASA"
promoter complement(7753..7830)
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
promoter 8064..8092
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 8100..8116
/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)."
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