Basic Vector Information
- Vector Name:
- pEF6/V5-His A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5818 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Invitrogen (Life Technologies)
- Copy Number:
- High copy number
- Promoter:
- EF-1α
pEF6/V5-His A 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/V5-His A vector Sequence
LOCUS 40924_17044 5818 bp DNA circular SYN 01-JAN-1980
DEFINITION Mammalian vector for high-level constitutive expression of
C-terminally V5- and 6xHis-tagged proteins. For other reading
frames, use pEF6/V5-His B or pEF6/V5-His C.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5818)
AUTHORS Invitrogen (Life Technologies)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5818)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5818
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 473..1651
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
promoter 1668..1686
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1805..1846
/codon_start=1
/label=V5 tag
/note="epitope tag from simian virus 5"
/translation="GKPIPNPLLGLDST"
CDS 1856..1873
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
polyA_signal 1902..2126
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2172..2600
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2614..2943
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 2991..3038
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 3057..3452
/codon_start=1
/label=BSD
/note="blasticidin S deaminase"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 3613..3746
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3783..3799)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3807..3823)
/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(3831..3861)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3876..3897)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4185..4773)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4947..5804)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(join(5805..5818,1..91))
/label=AmpR promoter
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