Basic Vector Information
- Vector Name:
- pEF6/V5-His B
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5811 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Invitrogen (Life Technologies)
- Copy Number:
- High copy number
- Promoter:
- EF-1α
pEF6/V5-His B 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 B vector Sequence
LOCUS 40924_17049 5811 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 A or pEF6/V5-His C. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5811) AUTHORS Invitrogen (Life Technologies) TITLE Direct Submission REFERENCE 2 (bases 1 to 5811) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..5811 /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 1809..1850 /codon_start=1 /label=V5 tag /note="epitope tag from simian virus 5" /translation="GKPIPNPLLGLDST" CDS 1860..1877 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" polyA_signal 1906..2130 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2176..2604 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2618..2947 /label=SV40 promoter /note="SV40 enhancer and early promoter" promoter 2995..3042 /label=EM7 promoter /note="synthetic bacterial promoter" CDS 3061..3456 /codon_start=1 /label=BSD /note="blasticidin S deaminase" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 3617..3750 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3787..3803) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3811..3827) /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(3835..3865) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3880..3901) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(4178..4766) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4940..5797) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(join(5798..5811,1..91)) /label=AmpR promoter
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