pEF1/myc-His A, B, and C are 6.2 kb vectors derived from pcDNA 3.1/myc-His and designed for overproduction of recombinant proteins in mammalian cell lines. Features of the vectors allow purification and detection of expressed proteins. High-level stable and transient expression can be carried out in most mammalian cells. The vectors contain the following elements:The control plasmid, pEF1/myc-His/lacZ, is included for use as a positive control for transfection, expression, and detection in the cell line of choice.
| 载体基本信息 | |||
| 载体名称 | pEF1/myc-His C | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 6161 bp | 载体类型 | Mammalian Expression Vectors |
| 筛选标记 | Neomycin | 拷贝数 | High copy number |
| 启动子 | EF-1a | 5引物 | T7 Fwd：5d[TAATACGACTCACTATAGGG]3 |
| 载体标签 | 6xHis, myc | ||
pEF1/myc-His C 载体载体图谱
pEF1/myc-His C 载体载体序列
LOCUS Exported 6161 bp ds-DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6161)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6161
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 473..1651
/note="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 704..1642
/note="EF-1-alpha intron A"
/note="intron upstream of the start codon of human
EF-1-alpha"
promoter 1668..1686
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1798..1827
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
CDS 1843..1860
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 1889..2113
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 2159..2587
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2601..2930
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2781..2916
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2997..3791
/codon_start=1
/gene="aph(3)-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3967..4088
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(4137..4153)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4161..4177
/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(4185..4215)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4230..4251
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4528..5116)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5287..6147)
/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(6148..91)
/gene="bla"
/note="AmpR promoter"
