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:
- pMQ351
- 载体长度:
- 9921 bp
- 载体类型:
- Cloning vector
- 复制子:
- R6K γ ori
- 宿主:
- Yeast
- 载体来源:
- Kalivoda EJ, Horzempa J, Stella NA, Sadaf A, Kowalski RP, Nau GJ, Shanks RM.
- 启动子:
- LEU2
pMQ351 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。
pMQ351 载体序列
LOCUS V004504 9921 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004504
VERSION V004504
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9921)
AUTHORS Kalivoda EJ, Horzempa J, Stella NA, Sadaf A, Kowalski RP, Nau GJ,
Shanks RM.
TITLE New vector tools with a hygromycin resistance marker for use with
opportunistic pathogens
JOURNAL Mol. Biotechnol. 48 (1), 7-14 (2011)
PUBMED 20972648
REFERENCE 2 (bases 1 to 9921)
AUTHORS Kalivoda EJ, Horzempa J, Sadaf A, Stella NA, Nau GJ, Shanks RM.
TITLE Direct Submission
JOURNAL Submitted (18-OCT-2010) Ophthalmology, University of Pittsburgh, 203
Lothrop St, Pittsburgh, PA 15213, USA
REFERENCE 3 (bases 1 to 9921)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9921)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
Biotechnol."; date: "2011"; volume: "48"; issue: "1"; pages: "7-14"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(18-OCT-2010) Ophthalmology, University of Pittsburgh, 203 Lothrop
St, Pittsburgh, PA 15213, USA"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9921
/mol_type="other DNA"
/organism="synthetic DNA construct"
oriT complement(399..508)
/direction=LEFT
/label="oriT"
/note="incP origin of transfer"
terminator complement(929..956)
/label="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(1048..1134)
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 1484..1500
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(1504..1560)
/label="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(1570..1586)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1594..1610
/label="lac operator"
/bound_moiety="lac repressor encoded by lacI"
/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(1618..1648)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(1663..1684)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin 1969..2357
/label="R6K gamma ori"
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(2892..3593)
/label="dTomato"
/note="dimeric variant of DsRed fluorescent protein (Shaner
et al., 2004)"
CDS 5380..5394
/label="enterokinase site"
/note="enterokinase recognition and cleavage site"
CDS complement(5635..6327)
/gene="SCEI"
/label="Intron-encoded endonuclease I-SceI"
/note="Intron-encoded endonuclease I-SceI from
Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
Accession#: P03882"
CDS complement(6355..7350)
/gene="hyg"
/label="Hygromycin-B 7''-O-kinase"
/note="Hygromycin-B 7''-O-kinase from Streptomyces
hygroscopicus. Accession#: P09979"
CDS complement(7639..8730)
/label="LEU2"
/note="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
promoter complement(8743..9147)
/label="LEU2 promoter"
misc_feature 9418..9921
/label="CEN/ARS"
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"