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:
- pMTL-JH30
- 载体长度:
- 3870 bp
- 载体类型:
- Integration vector
- 复制子:
- ori
- 载体来源:
- Heap JT, Ehsaan M, Cooksley CM, Ng YK, Cartman ST, Winzer K, Minton NP.
pMTL-JH30 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。
pMTL-JH30 载体序列
LOCUS V004430 3870 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004430
VERSION V004430
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 3870)
AUTHORS Heap JT, Ehsaan M, Cooksley CM, Ng YK, Cartman ST, Winzer K, Minton
NP.
TITLE Integration of DNA into bacterial chromosomes from plasmids without
a counter-selection marker
JOURNAL Nucleic Acids Res. 40 (8), E59 (2012)
PUBMED 22259038
REFERENCE 2 (bases 1 to 3870)
AUTHORS Heap JT, Ehsaan M, Cooksley CM, Cartman ST, Minton NP.
TITLE Direct Submission
JOURNAL Submitted (11-JAN-2011) School of Molecular Medical Sciences, The
University of Nottingham, Centre for Biomolecular Sciences,
University Park, Nottingham, Nottinghamshire NG7 2RD, United Kingdom
REFERENCE 3 (bases 1 to 3870)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3870)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2012"; volume: "40"; issue: "8"; pages: "E59"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(11-JAN-2011) School of Molecular Medical Sciences, The University
of Nottingham, Centre for Biomolecular Sciences, University Park,
Nottingham, Nottinghamshire NG7 2RD, United Kingdom"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3870
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 9..62
/label="derived from CD0164 gene of Clostridium difficile"
/note="derived from CD0164 gene of Clostridium difficile"
/regulatory_class="terminator"
primer_bind 63..79
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 90..389
/gene="thl fragment"
/note="mediates homologous recombination with the
chromosome of Clostridium acetobutylicum ATCC824"
gene 90..389
/gene="thl fragment"
/label="thl fragment"
CDS 412..987
/codon_start=1
/gene="pyrE"
/product="orotate phosphoribosyltransferase"
/label="pyrE"
/protein_id="AFA34992.1"
/translation="MSNINVIDILKESNALLEGHFLLSSGRHSNRYCQCAKLLQYPQKA
EKVISVVAEKLKDADFNIIVGPAMGGVIVSYELARQTNRPGIFAERKDGIMCIRRGFEI
KKGDKVIISEDVITTGKSSLEVAKVIEEMGGEVVGIACIVDRRAEGIKTNYPIYSACKL
EIETYEKDNCELCKKNIPFVKPGSREQK"
gene 412..987
/gene="pyrE"
/label="pyrE"
primer_bind complement(1106..1122)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
regulatory 1270..1311
/label="derived from fdx gene of Clostridium pasteurianum"
/note="derived from fdx gene of Clostridium pasteurianum"
/regulatory_class="terminator"
CDS 1690..2130
/codon_start=1
/gene="repL"
/product="plasmid replication protein"
/label="repL"
/protein_id="AFA34994.1"
/translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
gene 1690..2130
/gene="repL"
/label="repL"
CDS 2280..2900
/gene="catP"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from Clostridium
perfringens. Accession#: P26826"
rep_origin 3080..3668
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"