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。

载体名称:
pMQ236
载体抗性:
Kanamycin
载体长度:
5997 bp
载体类型:
Allelic replacement vector
复制子:
R6K γ ori
宿主:
Yeast
载体来源:
Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
启动子:
URA3

pMQ236 载体图谱

pMQ2365997 bp60012001800240030003600420048005400oriTrrnB T2 terminatorrrnB T1 terminatorM13 fwdMCSM13 revlac operatorlac promoterCAP binding siteT7 promoterR6K gamma oriNeoR/KanRI-SceI meganuclease siteSmall ribosomal subunit protein uS12URA3URA3 promoterCEN/ARS

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。

pMQ236 载体序列

LOCUS       V004511                 5997 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004511
VERSION     V004511
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5997)
  AUTHORS   Shanks RM, Kadouri DE, MacEachran DP, O'Toole GA.
  TITLE     New yeast recombineering tools for bacteria
  JOURNAL   Plasmid 62 (2), 88-97 (2009)
   PUBMED   19477196
REFERENCE   2  (bases 1 to 5997)
  AUTHORS   Shanks RMQ., Kadouri DE, MacEachran DP, O'Toole GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-MAY-2009) Ophthalmology, University of Pittsburgh, 203
            Lothrop St, Pittsburgh, PA 15213, USA
REFERENCE   3  (bases 1 to 5997)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5997)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
            date: "2009"; volume: "62"; issue: "2"; pages: "88-97"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (14-MAY-2009) Ophthalmology, University of Pittsburgh, 203 Lothrop
            St, Pittsburgh, PA 15213, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(352..461)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     terminator      complement(882..909)
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1001..1087)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     1437..1453
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    complement(1457..1513)
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1523..1539)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(1547..1563)
                     /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(1571..1601)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1616..1637)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        complement(1688..1706)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      1914..2302
                     /label="R6K gamma ori"
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             2653..3444
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     misc_feature    3463..3492
                     /label="I-SceI meganuclease site"
                     /note="I-SceI meganuclease site"
     CDS             3761..4132
                     /gene="rpsL"
                     /label="Small ribosomal subunit protein uS12"
                     /note="Small ribosomal subunit protein uS12 from Salmonella
                     paratyphi A (strain ATCC 9150 / SARB42). Accession#:
                     Q5PIW1"
     CDS             complement(4398..5198)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(5199..5419)
                     /label="URA3 promoter"
     misc_feature    5447..5950
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"