Basic Vector Information
All pEF or pUB vectors contain a strong promoter for high-level expression in mammalian cells, a choice of selection marker for generating stable cell lines, and an epitope tag for easy detection with a monoclonal antibody and rapid purification on nickel-chelating resin. Each vector is available in three reading frames to simplify cloning in-frame with the fusion tag.
- Vector Name:
- pUB6/V5-His C
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5459 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Invitrogen
- Selection Marker:
- Blasticidin
- Copy Number:
- High copy number
- Promoter:
- UbC
- Cloning Method:
- Enzyme Cut
- 5' Primer:
- T7 Fwd:5'd[TAATACGACTCACTATAGGG]3'
- Fusion Tag:
- His Tag (6x), V5 Epitope Tag
- Expression Method:
- Constiutive, Stable / Transient
pUB6/V5-His C 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 ℃.
pUB6/V5-His C vector Sequence
LOCUS 40924_44849 5459 bp DNA circular SYN 18-SEP-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5459) TITLE Direct Submission REFERENCE 2 (bases 1 to 5459) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..5459 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 18..1227 /label=UbC promoter /note="human ubiquitin C promoter" CDS 1323..1364 /codon_start=1 /label=V5 tag /note="epitope tag from simian virus 5" /translation="GKPIPNPLLGLDST" CDS 1374..1391 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" polyA_signal 1420..1644 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1690..2118 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2132..2462 /label=SV40 promoter /note="SV40 enhancer and early promoter" promoter 2510..2557 /label=EM7 promoter /note="synthetic bacterial promoter" CDS 2576..2971 /codon_start=1 /label=BSD /note="blasticidin S deaminase" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 3132..3265 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3302..3318) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3326..3342) /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(3350..3380) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3395..3416) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(3704..4292) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4466..5323) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5324..5428) /label=AmpR promoter
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