pFM46 载体图谱和序列

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:: pFM46

载体抗性:: Kanamycin

载体长度:: 3669 bp

载体类型:: Cloning vector

复制子:: p15A ori

载体来源:: Anderson JC, Voigt CA, Arkin AP.

pFM46 载体图谱

复制序列 NovoBuilder中查看图谱

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。

pFM46 载体序列

下载GenBank文件(.gb)

LOCUS       40924_20276        3669 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFM46, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3669)
  AUTHORS   Anderson JC, Voigt CA, Arkin AP.
  TITLE     Environmental signal integration by a modular AND gate
  JOURNAL   Mol. Syst. Biol. 3, 133 (2007)
  PUBMED    17700541
REFERENCE   2  (bases 1 to 3669)
  AUTHORS   Salis HM, Mirsky EA, Voigt CA.
  TITLE     Automated design of synthetic ribosome binding sites to control 
            protein expression
  JOURNAL   Nat. Biotechnol. 27 (10), 946-950 (2009)
  PUBMED    19801975
REFERENCE   3  (bases 1 to 3669)
  AUTHORS   Tamsir A, Tabor JJ, Voigt CA.
  TITLE     Robust multicellular computing using genetically encoded NOR gates 
            and chemical 'wires'
  JOURNAL   Nature 469 (7329), 212-215 (2011)
  PUBMED    21150903
REFERENCE   4  (bases 1 to 3669)
  AUTHORS   Moser F, Voigt CA.
  TITLE     Genetic Circuit Performance under Conditions Relevant for Industrial
            Bioreactors
  JOURNAL   Unpublished
REFERENCE   5  (bases 1 to 3669)
  AUTHORS   Anderson JC, Moser F, Tamsir A, Salis H.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JAN-2012) Biological Engineering, MIT, 500 Tech 
            Square, Rm 209D, Cambridge, MA 02139, USA
REFERENCE   6  (bases 1 to 3669)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 3669)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Syst. 
            Biol. 3, 133 (2007)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Nat. 
            Biotechnol."; date: "2009"; volume: "27"; issue: "10"; pages: 
            "946-950"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Nature"; 
            date: "2011"; volume: "469"; issue: "7329"; pages: "212-215"
COMMENT     SGRef: number: 4; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 5; type: "Journal Article"; journalName: "Submitted 
            (06-JAN-2012) Biological Engineering, MIT, 500 Tech Square, Rm 209D,
            Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 6; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(413..957)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     terminator      complement(1447..1490)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1493..1514
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     regulatory      1515..1549
                     /label=BBa_J23102
                     /note="BBa_J23102"
                     /regulatory_class="promoter"
     regulatory      1558..1570
                     /label=BBa_B0032
                     /note="BBa_B0032"
                     /regulatory_class="ribosome_binding_site"
     CDS             1577..2290
                     /label=yeGFP
                     /note="yeast-enhanced green fluorescent protein"
     terminator      2313..2384
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2400..2427
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    2434..2454
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2455..2514
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     CDS             complement(2675..3487)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"

pFM46 载体 (V006216)

pFM46 载体图谱

质粒操作方法

pFM46 载体序列