我们所展示的质粒图谱主要是从文献和开放数据库中收集而来，主要是为了方便研究工作，其中一小部分质粒进行了质量控制，可供科学家使用。
所有的产品都严格仅供科学研究使用，不能应用于临床试验，包括人体摄入、注射或外用的药理学用途。
确保质粒的关键元件正确，但是我们并不能保证实验结果。页面展示的图谱序列为理论序列，可能与测序结果不一致，请自行比对后确定是否满足要求。(如果测过序，本页面一般会提供下载)
开放数据库中的大多数载体序列都没有被完全测序。如果实际序列与参考序列的相似度超过99%，则将其视为正确。
由于科学研究是在探索未知，具有很大的不确定性，在任何情况下，我们都不承担超出质粒本身的额外经济损失或责任。
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
- 载体名称:
- pDisplay
- 载体抗性:
- Ampicillin
- 载体长度:
- 5324 bp
- 载体类型:
- Mammalian Surface Display System
- 复制子:
- ori
- 筛选标记:
- Neomycin
- 启动子:
- SV40
- 5'引物:
- T7-F:TAATACGACTCACTATAGGG
- 载体标签:
- HA Tag, c-Myc Epitope Tag, PDGFR Transmembrane Domain, IgK Leader Sequence
pDisplay 载体图谱
质粒操作方法
1. 发货形式：质粒干粉(常温运输，存于-20度，请务必先转化提质粒后使用)
2. 收到质粒干粉后请先5000rpm离心1min，再加入20μl ddH2O溶解质粒;（质粒复测的浓度有时候与标称值差距较大，这可能是因为冻干质粒在管中的位置、复溶效率、测量偏差以及管壁的吸附导致，因此建议先转化提质粒后再使用）
3. 取1支100μl 感受态于冰上解冻10min，加入2μl质粒，再冰浴30min后，42℃热激60s，不要搅动，再冰浴2min；
4. 加入900μl无抗的LB液体培养基，180rpm震荡37℃培养45min (30℃培养1-1.5小时);
5. 6000rpm离心5min，仅留100μl上清液重悬细菌沉淀，并涂布至目标质粒抗性的LB平板上;
6. 将平板倒置37℃培养14h，如果要求是30℃则培养20h; (菌落过多则将质粒稀释后再转化；没有菌落则加入10μl质粒转化；建议不要直接转表达感受态， 要先转克隆感受态，重提质粒后再导入表达感受态);
7. 挑取单菌落至LB液体培养基中，加入对应抗生素，220rpm震荡培养14h，根据实验需要和质粒提取试剂盒说明书提取质粒。
pDisplay 载体序列
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"