Basic Vector Information
- Vector Name:
- pEF6A-E-PKR-ND
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6582 bp
- Type:
- Mammalian expression vector
- Replication origin:
- ori
- Source/Author:
- De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R.
- Promoter:
- EF-1α
pEF6A-E-PKR-ND 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 ℃.
pEF6A-E-PKR-ND vector Sequence
LOCUS 40924_17059 6582 bp DNA circular SYN 17-DEC-2018
DEFINITION Mammalian expression vector pEF6A-E-PKR-ND, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6582)
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JOURNAL Unpublished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 6582)
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 6582)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6582)
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"Unpublished"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..6582
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 351..1529
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
promoter 1546..1564
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 1639..1677
/label=E-tag
/note="E-tag"
CDS 1687..2442
/codon_start=1
/note="unnamed protein product; hPKR-ND"
/protein_id="SJL86758.1"
/translation="MAGDLSAGFFMEELNTYRQKQGVVLKYQELPNSGPPHDRRFTFQV
IIDGREFPEGEGRSKKEAKNAAAKLAVEILNKEKKAVSPLLLTTTNSSEGLSMGNYIGL
INRIAQKKRLTVNYEQCASGVHGPEGFHYKCKMGQKEYSIGTGSTKQEAKQLAAKLAYL
QILSEETSVKSDYLSSGSFATTCESQSNSLVTSTLASESSSEGDFSADTSEINSNSDSL
NSSSLLMNGLRNNQRKAKRSLAPRFDLPD"
CDS 2464..2493
/label=Myc
/note="Myc (human c-Myc proto-oncogene) epitope tag"
CDS 2509..2526
/label=6xHis
/note="6xHis affinity tag"
polyA_signal 2555..2779
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2825..3253
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3267..3596
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 3644..3691
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 3710..4105
/label=BSD
/note="blasticidin S deaminase"
polyA_signal 4266..4399
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4436..4452)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4460..4476)
/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(4484..4514)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4529..4550)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4827..5415)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5589..6446)
/label=AmpR
/note="beta-lactamase"
promoter complement(6447..6551)
/label=AmpR promoter
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