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

pPLEX-50211048 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000RB T-DNA repeatoriVspectinomycin resistant proteinoritrfAIS1oriTLB T-DNA repeatS1 promoter derived from subterranean clover stunt virus DNA segment 1kanamycin resistanceS3 terminator derived from subterranean clover stunt virus DNA segment 3

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"