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:
pHQMini35S-GUS
载体抗性:
Ampicillin
载体长度:
5125 bp
载体类型:
Expression vector
复制子:
ori
载体来源:
Ma JZ, Ma YL, Huang F.

pHQMini35S-GUS 载体图谱

pHQMini35S-GUS5125 bp6001200180024003000360042004800oriAmpRAmpR promoterM13 fwdmini-35S(-87/+25) promoterGUS6xHisNOS terminatorM13 revlac operatorlac promoterCAP binding site

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。

pHQMini35S-GUS 载体序列

LOCUS       40924_24842        5125 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pHQMini35S-GUS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5125)
  AUTHORS   Ma JZ, Ma YL, Huang F.
  TITLE     An efficient plant transient expression vector system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5125)
  AUTHORS   Ma JZ, Ma YL, Huang F.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JAN-2014) School of Life Science and Engineering, 
            LanZhou University of Technology, No.287 Langongping Road, 
            QiliheDistrict, Lanzhou 730050, China
REFERENCE   3  (bases 1 to 5125)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5125)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (22-JAN-2014) School of Life Science and Engineering, LanZhou 
            University of Technology, No.287 Langongping Road, QiliheDistrict, 
            Lanzhou 730050, China"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(945..1802)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1803..1907)
                     /label=AmpR promoter
     primer_bind     2381..2397
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2431..2543
                     /label=mini-35S(-87/+25) promoter
                     /note="mini-35S(-87/+25) promoter"
                     /regulatory_class="promoter"
     promoter        2486..2532
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     intron          2566..2755
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     CDS             2750..4573
                     /label=GUS
                     /note="beta-glucuronidase"
     CDS             4580..4597
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      4632..4884
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4906..4922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4930..4946)
                     /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(4954..4984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4999..5020)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."