首页技术支持质粒载体

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。

pNV18 is created by inserting a 1.8 kb DNA fragment containing the pAL5000 origin of replication into the unique NheI site of either pK18. Key features of pNV18 include: 1. Compact size. 2. Multiple cloning sites: The latest version includes sites for HindIII, SphI, PstI, SalI, HincII, XbaI, BamHI, KpnI, and EcoRI. 3. The lacZ gene (α fragment) facilitates blue-white selection in E. coli. 4. The aph gene from Tn5 provides kanamycin and neomycin resistance in E. coli and Nocardia species. 5. Broad host range: Effective in N. farcinica, N. asteroides, and various mycobacteria.

载体名称:
pNV18
载体抗性:
Kanamycin
载体长度:
4411 bp
载体类型:
Shuttle vector
复制子:
ori
载体来源:
Chiba K, Hoshino Y, Ishino K, Kogure T, Mikami Y, Uehara Y, Ishikawa J.
感受态:
Top10
培养温度:
37℃

pNV18 载体图谱

pNV184411 bp600120018002400300036004200M13 fwdMCSM13 revlac operatorlac promoterCAP binding siteoriNeoR/KanRderived from pAL5000

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。

pNV18 载体序列

LOCUS       40924_33652        4411 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNV18 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4411)
  AUTHORS   Chiba K, Hoshino Y, Ishino K, Kogure T, Mikami Y, Uehara Y, Ishikawa
            J.
  TITLE     Construction of a pair of practical Nocardia-Escherichia coli 
            shuttle vectors
  JOURNAL   Jpn. J. Infect. Dis. 60 (1), 45-47 (2007)
  PUBMED    17314425
REFERENCE   2  (bases 1 to 4411)
  AUTHORS   Ishikawa J, Chiba K.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JUL-2006) Contact:Jun Ishikawa National Institute of 
            Infectious Diseases, Department of Bioactive Molecules; 1-23-1 
            Toyama, Shinjuku, Tokyo 162-8640, Japan URL 
            :http://nocardia.nih.go.jp/
REFERENCE   3  (bases 1 to 4411)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4411)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Jpn. J. 
            Infect. Dis."; date: "2007"; volume: "60"; issue: "1"; pages: 
            "45-47"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-JUL-2006) Contact:Jun Ishikawa National Institute of Infectious 
            Diseases, Department of Bioactive Molecules; 1-23-1 Toyama, 
            Shinjuku, Tokyo 162-8640, Japan URL :http://nocardia.nih.go.jp/"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     On Dec 18, 2007 this sequence version replaced AB267085.1.
FEATURES             Location/Qualifiers
     source          1..4411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..168
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(172..228)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(238..254)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(262..278)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(286..316)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(331..352)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(640..1228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1515..2306)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    2662..4411
                     /label=derived from pAL5000
                     /note="derived from pAL5000"
     CDS             complement(2746..3093)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein RepB"
                     /label=repB
                     /protein_id="BAF02356.2"
                     /translation="MSDGYSDGYNRQPTVRKKRRVTAAEGARITGLSERHVVRLVAQER
                     SEWLAEQAARRERIRAYHDDEGHSWPQTAKHFGLHLDTVKRLGYRARKERAAEQEAAQK
                     AHNEADNPPLF"
     gene            complement(2746..3093)
                     /gene="repB"
                     /label=repB
     CDS             complement(3090..4016)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein RepA"
                     /label=repA
                     /protein_id="BAF02357.1"
                     /translation="MSHVADEFEQLWLPYWPLASDDLLEGIYRQSRASALGRRYIEANP
                     TALANLLVVDVDHPDAALRALSARGSHPLPNAIVGNRANGHAHAVWALNAPVPRTEYAR
                     RKPLAYMAACAEGLRRAVDGDRSYSGLMTKNPGHIAWETEWLHSDLYTLSHIEAELGAN
                     MPPPRWRQQTTYKAAPTPLGRNCALFDSVRLWAYRPALMRIYLPTRNVDGLGRAIYAEC
                     HARNAEFPCNDVCPGPLPDSEVRAIANSIWRWITTKSRIWADGIVVYEATLSARQSAIS
                     RKGAAARTAASTVARRAKSASAMEALL"
     gene            complement(3090..4016)
                     /gene="repA"
                     /label=repA