pPR9TT 载体 (V003932)

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:
pPR9TT
载体抗性:
Ampicillin
载体长度:
9388 bp
载体类型:
Promoter probe vector
复制子:
oriV
载体来源:
Santos PM, Blatny JM, Di Bartolo I, Valla S, Zennaro E.

pPR9TT 载体图谱

pPR9TT9388 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200oriTAmpR promoterAmpRoriVtrfAM13 fwdCmRbidirectionalSK primerlacZKS primerrrnB T2 terminatorrrnB T1 terminator

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。

pPR9TT 载体序列

LOCUS       40924_35454        9388 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Promoter probe vector pPR9TT, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9388)
  AUTHORS   Santos PM, Blatny JM, Di Bartolo I, Valla S, Zennaro E.
  TITLE     Physiological analysis of the expression of the styrene degradation 
            gene cluster in Pseudomonas fluorescens ST
  JOURNAL   Appl. Environ. Microbiol. 66 (4), 1305-1310 (2000)
  PUBMED    10742204
REFERENCE   2  (bases 1 to 9388)
  AUTHORS   Santos PM, Di Bartolo I, Blatny JM, Zennaro E, Valla S.
  TITLE     New broad-host-range promoter probe vectors based on the plasmid RK2
            replicon
  JOURNAL   FEMS Microbiol. Lett. 195 (1), 91-96 (2001)
  PUBMED    11167001
REFERENCE   3  (bases 1 to 9388)
  AUTHORS   Santos PM, Blatny JM, Di Bartolo I, Valla S, Zennaro E.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-SEP-1999) Department of Biology, Third University of 
            Rome, Viale Marconi 446, Rome 00146, Italy
REFERENCE   4  (bases 1 to 9388)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 9388)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2000"; volume: "66"; issue: "4"; pages:
            "1305-1310"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "FEMS 
            Microbiol. Lett."; date: "2001"; volume: "195"; issue: "1"; pages: 
            "91-96"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (16-SEP-1999) Department of Biology, Third University of Rome, Viale
            Marconi 446, Rome 00146, Italy"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(67..175)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        352..456
                     /label=AmpR promoter
     CDS             457..1314
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1564..2094
                     /label=oriV
                     /note="incP origin of replication"
     regulatory      2316..2496
                     /label=Pneo
                     /note="Pneo"
                     /regulatory_class="promoter"
     CDS             2486..3631
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     primer_bind     complement(3815..3831)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4327..4983
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     regulatory      5150..5724
                     /label=bidirectional
                     /note="bidirectional"
                     /regulatory_class="terminator"
     primer_bind     5775..5791
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5813..8857)
                     /label=lacZ
                     /note="beta-galactosidase"
     primer_bind     complement(8897..8913)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(8964..8991)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(9083..9169)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"