pDisplay is a mammalian expression vector designed to target recombinant proteins to the surface of mammalian cells. Displayed proteins can be analyzed for their ability to interact with known or putative ligands. Proteins of interest are targeted and anchored to the cell surface by cloning the gene of interest in frame with the vectors unique N-terminal secretion signal and the C-terminal transmembrane anchoring domain of platelet-derived growth factor receptor (PDGFR). In contrast to phage display vectors which operate exclusively in prokaryotic cells, the pDisplay vector offers the advantage of having the displayed protein of interest processed in mammalian cells. Therefore, recombinant proteins of eukaryotic origin that are expressed from pDisplay more closely resemble their native form. This may favor more accurate ligand binding interactions. In addition to the N-terminal cell surface targeting signal and C-terminal transmembrane anchoring domain, the pDisplay vector contains the following key features:• T7 promoter/priming site for in vitro transcription of sense RNA and for sequencing of inserts • Neomycin resistance marker for stable selection in mammalian cells using Geneticin • SV40 origin for replication and simple vector rescue in cell lines expressing the large T antigen (e.g., COS-1 and COS-7) • Ampicillin resistance gene for selection in E. coli
| 载体基本信息 | |||
| 载体名称 | pDisplay | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 5325 bp | 载体类型 | Mammalian Surface Display System |
| 筛选标记 | Neomycin | 启动子 | CMV |
| 5引物 | T7-F:TAATACGACTCACTATAGGG | 载体标签 | HA Tag, c-Myc Epitope Tag, PDGFR Transmembrane Domain, IgK Leader Sequence |
pDisplay 载体载体图谱
pDisplay 载体载体序列
LOCUS Exported 5325 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 5325)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5325
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 10..389
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 390..593
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 638..656
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 737..799
/codon_start=1
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/note="Ig-kappa leader"
/translation="METDTLLLWVLLLWVPGSTGD"
CDS 800..826
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/note="HA"
/translation="YPYDVPDYA"
CDS 874..903
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/note="Myc"
/translation="EQKLISEEDL"
polyA_signal 1081..1305
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(1437..2025)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
polyA_signal 2354..2401
/note="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
promoter complement(3456..3816)
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 3470..3608
/note="SV40 ori"
/note="SV40 origin of replication"
CDS complement(3876..4736)
/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(4737..4841)
/gene="bla"
/note="AmpR promoter"
rep_origin 4868..5323
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
