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.
| 载体基本信息 | |||
| 载体名称 | pUB6/V5-His C | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 5459 bp | 载体类型 | Mammalian Expression Vectors |
| 载体来源 | Invitrogen | 筛选标记 | Blasticidin |
| 拷贝数 | High copy number | 启动子 | UbC promoter |
| 克隆方法 | Enzyme Cut | 5引物 | T7 Fwd:5'd[TAATACGACTCACTATAGGG]3' |
| 载体标签 | His Tag (6x), V5 Epitope Tag | 表达方法 | Constiutive, Stable / Transient |
pUB6/V5-His C 载体载体图谱
pUB6/V5-His C 载体载体序列
LOCUS Exported 5459 bp ds-DNA circular SYN 18-SEP-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5459)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5459
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 18..1227
/note="UbC promoter"
/note="human ubiquitin C promoter"
CDS 1323..1364
/codon_start=1
/product="epitope tag from simian virus 5"
/note="V5 tag"
/translation="GKPIPNPLLGLDST"
CDS 1374..1391
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 1420..1644
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 1690..2118
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2132..2462
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2313..2448
/note="SV40 ori"
/note="SV40 origin of replication"
promoter 2510..2557
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 2576..2974
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/note="BSD"
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 3132..3253
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(3302..3318)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3326..3342
/bound_moiety="lac repressor encoded by lacI"
/note="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)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3395..3416
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3704..4292)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4463..5323)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5324..5428)
/gene="bla"
/note="AmpR promoter"
