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:
- pMA_Level1D
- 载体抗性:
- Kanamycin
- 载体长度:
- 3101 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Andreou AI, Nakayama N.
pMA_Level1D 载体图谱
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。
pMA_Level1D 载体序列
LOCUS 40924_29856 3101 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMA_Level1D, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3101)
AUTHORS Andreou AI, Nakayama N.
TITLE Mobius Assembly: A versatile Golden-Gate framework towards universal
DNA assembly
JOURNAL PLoS ONE 13 (1), e0189892 (2018)
PUBMED 29293531
REFERENCE 2 (bases 1 to 3101)
AUTHORS Andreou AI, Nakayama N.
TITLE Direct Submission
JOURNAL Submitted (20-OCT-2017) Centre for Synthetic and Systems Biology and
Institute of Molecular Plant Sciences, University of Edinburgh, Max
Born Crescent, Kings Buildings, Edinburgh EH9 3BF, UK
REFERENCE 3 (bases 1 to 3101)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3101)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
date: "2018"; volume: "13"; issue: "1"; pages: "e0189892"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(20-OCT-2017) Centre for Synthetic and Systems Biology and Institute
of Molecular Plant Sciences, University of Edinburgh, Max Born
Crescent, Kings Buildings, Edinburgh EH9 3BF, UK"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3101
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 2..59
/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."
rep_origin complement(254..842)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator complement(1025..1119)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(1143..1955)
/label=KanR
/note="aminoglycoside phosphotransferase"
promoter complement(1956..2059)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
terminator complement(2139..2182)
/label=bacterial terminator
/note="putative bacterial transcription terminator"
misc_feature 2202..2205
/label=CTTG overlap
/note="CTTG overlap"
misc_feature 2206..2209
/label=GGAG overlap
/note="GGAG overlap"
regulatory 2217..2251
/label=Anderson J23106 promoter
/note="Anderson J23106 promoter"
/regulatory_class="promoter"
regulatory 2260..2271
/label=B0034 RBS
/note="B0034 RBS"
/regulatory_class="ribosome_binding_site"
RBS 2260..2271
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 2278..2949
/label=spisCP
/note="spisCP is a fluorescent protein published in 2008,
derived from Stylophora pistillata. It is reported to be a
tetramer."
terminator 2960..3031
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3047..3074
/label=T7Te terminator
/note="phage T7 early transcription terminator"
misc_feature 3082..3085
/label=CGCT overlap
/note="CGCT overlap"