pQLinkG 载体 (V003853)

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:
pQLinkG
载体抗性:
Ampicillin
载体长度:
5537 bp
载体类型:
Cloning vector
复制子:
ori
载体来源:
Scheich C, Kummel D, Soumailakakis D, Heinemann U, Bussow K.

pQLinkG 载体载体图谱

pQLinkG5537 bp60012001800240030003600420048005400LIC cassette with PacI restriction siteT5 promoterlac operatorRBSGSTattB1TEV siteattB2lambda t0 terminatorLIC cassette with SwaI, PacI restriction sitesCmRrrnB T1 terminatorCAP binding sitelacIlacIq promoterbomoriAmpRAmpR promoter

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。

pQLinkG 载体载体序列

LOCUS       40924_36208        5537 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pQLinkG, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5537)
  AUTHORS   Scheich C, Kummel D, Soumailakakis D, Heinemann U, Bussow K.
  TITLE     Vectors for co-expression of an unrestricted number of proteins
  JOURNAL   Nucleic Acids Res. 35 (6), E43 (2007)
  PUBMED    17311810
REFERENCE   2  (bases 1 to 5537)
  AUTHORS   Buessow K, Scheich C.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-SEP-2006) Vertebrate Genomics, Max Planck Institute 
            for Molecular Genetics, Ihnestr. 63-73, Berlin 14195, Germany
REFERENCE   3  (bases 1 to 5537)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5537)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2007"; volume: "35"; issue: "6"; pages: "E43"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-SEP-2006) Vertebrate Genomics, Max Planck Institute for 
            Molecular Genetics, Ihnestr. 63-73, Berlin 14195, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..27
                     /label=LIC cassette with PacI restriction site
                     /note="LIC cassette with PacI restriction site"
     promoter        36..80
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA
                     polymerase, with embedded lac operator"
     protein_bind    88..104
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     RBS             123..134
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      127..133
                     /regulatory_class="ribosome_binding_site"
     CDS             141..794
                     /label=GST
                     /note="glutathione S-transferase from Schistosoma
                     japonicum"
     protein_bind    817..837
                     /label=attB1
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     CDS             840..860
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     protein_bind    complement(898..922)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      944..1038
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1043..1092
                     /label=LIC cassette with SwaI, PacI restriction sites
                     /note="LIC cassette with SwaI, PacI restriction sites"
     CDS             1132..1788
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      1856..1942
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     protein_bind    complement(2001..2022)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2038..3117)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3118..3195)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    3387..3527
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      3711
                     /label=ColE1 ori
                     /note="ColE1 ori"
     rep_origin      complement(3713..4301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4475..5332)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5333..5437)
                     /label=AmpR promoter