pDisplay vector (V000792)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V000792	pDisplay	In stock, instant shipping	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

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

Vector Name:: pDisplay

Antibiotic Resistance:: Ampicillin

Length:: 5324 bp

Type:: Mammalian Surface Display System

Replication origin:: ori

Selection Marker:: Neomycin

Promoter:: SV40

5' Primer:: T7-F:TAATACGACTCACTATAGGG

Fusion Tag:: HA Tag, c-Myc Epitope Tag, PDGFR Transmembrane Domain, IgK Leader Sequence

pDisplay vector Map

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Plasmid Protocol

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5. Store the plasmid at -20 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

References

Wang J, Li X, Liu C, Wang S, Li J. Effect of BCG HSP70 Gene Transfection on Dendritic Cells Derived From Bone Marrow in Children With Acute Leukemia. J Pediatr Hematol Oncol. 2022 Aug 1;44(6):e939-e944.

pDisplay vector Sequence

LOCUS       Exported                5324 bp DNA     circular SYN 17-OCT-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5324)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5324)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5324)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2950..2955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     737..799
                     /label=Ig-kappa leader
                     /note="leader sequence from mouse immunoglobulin kappa
                     light
                     chain"
     CDS             800..826
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             874..903
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             907..1053
                     /codon_start=1
                     /label=PDGFR-beta TM domain
                     /note="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     polyA_signal    1081..1305
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1437..2025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    complement(2354..2401)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(2636..3427)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(3462..3815)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             complement(3878..4735)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4736..4840)
                     /label=AmpR promoter
     rep_origin      4867..5322
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"