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。

The pME6031 is a broad-host vector for studies requiring multi-color tagging and visualization of plant-associated, Gram-negative bacterial strains.

载体名称:
pME6031
载体抗性:
Tetracycline
载体长度:
8310 bp
载体类型:
Shuttle vector
复制子:
p15A ori
载体来源:
Heeb S, Itoh Y, Nishijyo T, Schnider U, Keel C, Wade J, Walsh U, O'Gara F, Haas D.
感受态:
JM108
培养温度:
37℃

pME6031 载体图谱

pME60318310 bp400800120016002000240028003200360040004400480052005600600064006800720076008000pVS1 resolvasepVS1 StaAhypothetical proteinKorB boxpVS1 RepApVS1 oriVp15A orip15A origin of transferpart of terminator absent in pME6030multiple cloning siteTetRTcR

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。

pME6031 载体序列

LOCUS       40924_30555        8310 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pME6031, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8310)
  AUTHORS   Heeb S, Itoh Y, Nishijyo T, Schnider U, Keel C, Wade J, Walsh U, 
            O'Gara F, Haas D.
  TITLE     Small, stable shuttle vectors based on the minimal pVS1 replicon for
            use in gram-negative, plant-associated bacteria
  JOURNAL   Mol. Plant Microbe Interact. 13 (2), 232-237 (2000)
  PUBMED    10659714
REFERENCE   2  (bases 1 to 8310)
  AUTHORS   Heeb S.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-DEC-1998) Laboratoire de Biologie Microbienne, 
            University of Lausanne, Batiment de Biologie, Lausanne CH-1015, 
            Switzerland
REFERENCE   3  (bases 1 to 8310)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8310)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Plant 
            Microbe Interact."; date: "2000"; volume: "13"; issue: "2"; pages: 
            "232-237"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-DEC-1998) Laboratoire de Biologie Microbienne, University of 
            Lausanne, Batiment de Biologie, Lausanne CH-1015, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             195..881
                     /codon_start=1
                     /product="pVS1 resolvase"
                     /label=pVS1 resolvase
                     /note="ORF1; not required for stable maintenance"
                     /protein_id="AAD19683.1"
                     /translation="MNKSAAAGLLGYARVSTDDQDLTNQRAELHAAGCTKLFSEKITGT
                     RRDRPELARMLDHLRPGDVVTVTRLDRLARSTRDLLDIAERIQEAGAGLRSLAEPWADT
                     TTPAGRMVLTVFAGIAEFERSLIIDRTRSGREAAKARGVKFGPRPTLTPAQIAHARELI
                     DQEGRTVKEAAALLGVHRSTLYRALERSEEVTPTEARRRGAFREDALTEADALAAAENE
                     RQEEQA"
     CDS             878..1093
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF2; not required for stable maintenance"
                     /protein_id="AAD19684.1"
                     /translation="MKPHQDGQDEPFFITEEIEAEMIAAGYVFEPPAHVSTVRLHEILA
                     GLSDAKLAAWPASLAAEETERRRLKR"
     CDS             1180..1806
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             1830..2045
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="ORF3; not required for stable maintenance"
                     /protein_id="AAD19686.1"
                     /translation="MSKSTNTLSAGRPSARSSKAATLASLADTPAMKRVNFQLPAEDHT
                     KLKMYAVRQGKTITELLSEYIAQLPE"
     misc_feature    2075..2088
                     /label=KorB box
                     /note="KorB box"
     CDS             2238..3308
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      3377..3571
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      4281..4826
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     misc_feature    5068..5089
                     /label=p15A origin of transfer
                     /note="p15A origin of transfer"
     misc_difference 5136..5414
                     /label=part of terminator absent in pME6030
                     /note="part of terminator absent in pME6030"
     regulatory      5148..5414
                     /label=T4 transcription terminator
                     /note="T4 transcription terminator"
                     /regulatory_class="terminator"
     misc_feature    5416..5526
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             complement(6318..6965)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     CDS             7071..8267
                     /label=TcR
                     /note="tetracycline efflux protein"