pVMG-TnpR 载体 (V002230)

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:
pVMG-TnpR
载体抗性:
Ampicillin
载体长度:
7264 bp
载体类型:
Cloning vector
复制子:
ori
载体来源:
Oke V, Long SR.

pVMG-TnpR 载体图谱

pVMG-TnpR7264 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200Escherichia coli trpAcontains stop codons in all three reading framesregulatoryTransposon gamma-delta resolvaseregulatoryGUSNeoR/KanRM13 fwdf1 orioriTAmpR promoterAmpRori

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。

pVMG-TnpR 载体序列

LOCUS       V002230                 7264 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V002230
VERSION     V002230
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7264)
  AUTHORS   Oke V, Long SR.
  TITLE     Bacterial genes induced within the nodule during the
            Rhizobium-legume symbiosis
  JOURNAL   Mol. Microbiol. 32 (4), 837-849 (1999)
   PUBMED   10361286
REFERENCE   2  (bases 1 to 7264)
  AUTHORS   Gao M, Teplitski M.
  TITLE     RIVET-a tool for in vivo analysis of symbiotically relevant gene
            expression in Sinorhizobium meliloti
  JOURNAL   Mol. Plant Microbe Interact. 21 (2), 162-170 (2008)
   PUBMED   18184060
REFERENCE   3  (bases 1 to 7264)
  AUTHORS   Gao M, Oke V, Teplitski M.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-OCT-2007) Soil and Water Science, University of
REFERENCE   4  (bases 1 to 7264)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7264)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
            Microbiol."; date: "1999"; volume: "32"; issue: "4"; pages:
            "837-849"
            SGRef: number: 2; type: "Journal Article"; journalName: "Mol. Plant
            Microbe Interact."; date: "2008"; volume: "21"; issue: "2"; pages:
            "162-170"
            SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (17-OCT-2007) Soil and Water Science, University of Florida-IFAS,
            1376 Mowry Rd., Cancer "
            SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      28..58
                     /label="Escherichia coli trpA"
                     /note="Escherichia coli trpA"
                     /regulatory_class="terminator"
     misc_feature    103..113
                     /label="contains stop codons in all three reading frames"
                     /note="contains stop codons in all three reading frames"
     regulatory      189..195
                     /gene="tnpR"
                     /regulatory_class="ribosome_binding_site"
     CDS             205..753
                     /gene="tnpR"
                     /label="Transposon gamma-delta resolvase"
                     /note="Transposon gamma-delta resolvase from Escherichia
                     coli (strain K12). Accession#: P03012"
     regulatory      756..761
                     /gene="uidA"
                     /regulatory_class="ribosome_binding_site"
     CDS             788..2596
                     /label="GUS"
                     /note="beta-glucuronidase"
     CDS             3019..3810
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     primer_bind     complement(4183..4199)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      4412..4867
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     oriT            complement(5206..5315)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     promoter        5485..5589
                     /label="AmpR promoter"
     CDS             5590..6447
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      6621..7209
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"