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pSecTag2A-E06-scFv vector

Basic Vector Information

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.

Basic Vector Information
Vector Name pSecTag2A-E06-scFv Antibiotic Resistance Ampicillin
Length 5846 bp Type Mammalian Expression Vectors
Source Dr. Joseph Witztum Selection Marker Zeocin
Copy Number High copy number Promoter CMV
Cloning Method Enzyme Cut 5' Primer T7 promoter
3' Primer BGH Reverse primer Fusion Tag Myc,6*His

pSecTag2A-E06-scFv vector Vector Map

pSecTag2A-E06-scFv vector Vector Map

pSecTag2A-E06-scFv vector Sequence

Copy Sequence

Download SnapGene Map(.dna)

Download GeneBank File(.gb)

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"

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