The E06 antibody is a murine monoclonal IgM natural antibody that binds to oxidized phospholipid and reacts with oxidized LDL, oxidized HDL and protein covalently modified by oxidized phospholipid. This antibody has been used to probe atherosclerotic plaque in mice, zebrafish, rabbits, monkeys and humans. It also binds to apoptotic cells, but not viable cells, and can be used in a variety of applications, including ELISA, immunohistochemistry and Western blot analysis, as well as for in vivo imaging with MRI techniques.
| 载体基本信息 | |||
| 载体名称 | pSecTag2A-E06-scFv | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 5846 bp | 载体类型 | Mammalian Expression Vectors |
| 载体来源 | Dr. Joseph Witztum | 筛选标记 | Zeocin |
| 拷贝数 | High copy number | 启动子 | CMV |
| 克隆方法 | Enzyme Cut | 5引物 | T7 promoter |
| 3引物 | BGH Reverse primer | 载体标签 | Myc,6*His |
pSecTag2A-E06-scFv 载体载体图谱
pSecTag2A-E06-scFv 载体载体序列
LOCUS Exported 5846 bp ds-DNA circular SYN 11-JUN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pSecTag2A-E06-scFv
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5846)
AUTHORS Triple Threat
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5846
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 905..967
/codon_start=1
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/note="Ig-kappa leader"
/translation="METDTLLLWVLLLWVPGSTGD"
CDS 968..1342
/codon_start=1
/note="E06 VL"
/translation="AAQPARRAVRSLDIVMTQSPSSLSVSAGKKVTISCTASESLYSSK
HKVHYLAWYQKKPEQSPKLLIYGASNRYIGVPDRFTGSGSGTDFTLTISSVQVEDLTHY
YCAQFYSYPLTFGAGTKLEIK"
CDS 1343..1387
/codon_start=1
/note="G4S x 3"
/translation="GGGGSGGGGSGGGGS"
CDS 1388..1768
/codon_start=1
/note="E06 VH"
/translation="EVKLVESGGGLVQPGGSLRLSCATSGFTFSDFYMEWVRQAPGKRL
EWIAASRNKANDYTTEYADSVKGRFIVSRDTSQSILYLQMNALRAEDTAIYYCARDYYG
SSYWYFDVWGAGTTVTVSSRGGP"
CDS 1769..1798
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
CDS 1814..1831
/codon_start=1
/product="6xHis affinity tag"
/note="6xHis"
/translation="HHHHHH"
polyA_signal 1860..2084
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 2130..2558
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2572..2901
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2752..2887
/note="SV40 ori"
/note="SV40 origin of replication"
promoter 2949..2996
/note="EM7 promoter"
/note="synthetic bacterial promoter "
CDS 3015..3389
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/note="BleoR"
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 3519..3640
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(3689..3705)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3713..3729
/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(3737..3767)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3782..3803
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4091..4679)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4850..5710)
/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(5711..5815)
/gene="bla"
/note="AmpR promoter"
