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。
- 载体名称:
- pPLEX-502
- 载体长度:
- 11048 bp
- 载体类型:
- Cloning vector
- 复制子:
- oriV
- 载体来源:
- Schunmann PHD., Llewellyn DJ, Surin B, Boevink P, De Feyter RC, Waterhouse PM.
pPLEX-502 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。
pPLEX-502 载体序列
LOCUS 40924_35024 11048 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pPLEX-502, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11048)
AUTHORS Schunmann PHD., Llewellyn DJ, Surin B, Boevink P, De Feyter RC,
Waterhouse PM.
TITLE A suite of novel promoters and terminators for plant biotechnology
JOURNAL Funct. Plant Biol. 30, 443-452 (2003)
REFERENCE 2 (bases 1 to 11048)
AUTHORS Schunmann PHD.
TITLE Direct Submission
JOURNAL Submitted (06-OCT-2002) Plant Industry, CSIRO, Clunies Ross Street,
Canberra, ACT 2602, Australia
REFERENCE 3 (bases 1 to 11048)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11048)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Funct.
Plant Biol. 30, 443-452 (2003)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-OCT-2002) Plant Industry, CSIRO, Clunies Ross Street, Canberra,
ACT 2602, Australia"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11048
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 168..192
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
rep_origin 672..1385
/label=oriV
/note="incP origin of replication"
CDS complement(1990..2778)
/codon_start=1
/product="spectinomycin resistant protein"
/label=spectinomycin resistant protein
/protein_id="AAP22319.1"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEDRLASRADQLEEFVHYVKGEITKVVGK"
rep_origin 3875..4463
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5826..6971)
/label=trfA
/note="trans-acting replication protein that binds to and
activates oriV"
CDS complement(7246..7596)
/label=traJ
/note="oriT-recognizing protein"
mobile_element complement(7596..8363)
/label=IS1
/note="prokaryotic transposable element"
oriT complement(8423..8532)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
misc_feature 9087..9111
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
regulatory 9191..9713
/note="S1 promoter derived from subterranean clover stunt
virus DNA segment 1"
/regulatory_class="promoter"
CDS join(9723..9899,10090..10707)
/codon_start=1
/product="neomycin phosphotransferase II"
/label=neomycin phosphotransferase II
/note="kanamycin resistance"
/protein_id="AAP21556.1"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
intron 9900..10089
/note="modified CAT-1"
intron 9906..10095
/label=cat1 intron
/note="castor bean catalase intron, modified"
regulatory 10889..11027
/note="S3 terminator derived from subterranean clover stunt
virus DNA segment 3"
/regulatory_class="terminator"