Basic Vector Information
- Vector Name:
- pEF6-FLAG-hPKR-ND
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6528 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
pEF6-FLAG-hPKR-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 ℃.
pEF6-FLAG-hPKR-ND vector Sequence
LOCUS 40924_17024 6528 bp DNA circular SYN 17-DEC-2018 DEFINITION Mammalian expression vector pEF6-FLAG-hPKR-ND, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6528) 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A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R. TITLE BCCM/LMBP Plasmid collection JOURNAL Unpublished REFERENCE 2 (bases 1 to 6528) AUTHORS De Schamphelaire W. TITLE Direct Submission JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark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 6528) TITLE Direct Submission REFERENCE 4 (bases 1 to 6528) 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 TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 9052, BELGIUM"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..6528 /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" CDS 1600..1623 /label=FLAG /note="FLAG(R) epitope tag, followed by an enterokinase cleavage site" CDS 1633..2388 /codon_start=1 /note="unnamed protein product; hPKR-ND" /protein_id="SJL86764.1" /translation="MAGDLSAGFFMEELNTYRQKQGVVLKYQELPNSGPPHDRRFTFQV IIDGREFPEGEGRSKKEAKNAAAKLAVEILNKEKKAVSPLLLTTTNSSEGLSMGNYIGL INRIAQKKRLTVNYEQCASGVHGPEGFHYKCKMGQKEYSIGTGSTKQEAKQLAAKLAYL QILSEETSVKSDYLSSGSFATTCESQSNSLVTSTLASESSSEGDFSADTSEINSNSDSL NSSSLLMNGLRNNQRKAKRSLAPRFDLPD" CDS 2410..2439 /label=Myc /note="Myc (human c-Myc proto-oncogene) epitope tag" CDS 2455..2472 /label=6xHis /note="6xHis affinity tag" polyA_signal 2501..2725 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2771..3199 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3213..3542 /label=SV40 promoter /note="SV40 enhancer and early promoter" promoter 3590..3637 /label=EM7 promoter /note="synthetic bacterial promoter" CDS 3656..4051 /label=BSD /note="blasticidin S deaminase" polyA_signal 4212..4345 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4382..4398) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(4406..4422) /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(4430..4460) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(4475..4496) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4773..5361) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5535..6392) /label=AmpR /note="beta-lactamase" promoter complement(6393..6497) /label=AmpR promoter
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