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:
- pOpt_Clover_Hyg
- 载体抗性:
- Ampicillin
- 载体长度:
- 6831 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Lauersen KJ, Mussgnug JH, Kruse O.
pOpt_Clover_Hyg 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。
pOpt_Clover_Hyg 载体序列
LOCUS V004221 6831 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004221
VERSION V004221
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6831)
AUTHORS Lauersen KJ, Mussgnug JH, Kruse O.
TITLE Efficient targeted expression of nuclear transgenes in Chlamydomonas
reinhardtii with a novel, modular vector toolkit
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6831)
AUTHORS Lauersen KJ, Mussgnug JH, Kruse O.
TITLE Direct Submission
JOURNAL Submitted (25-JUN-2014) Algae Biotechnology and Bioenergy,
University of Bielefeld Center for Biotechnology, Universitaetsstr
27, Bielefeld, NRW 33615, Germany
REFERENCE 3 (bases 1 to 6831)
AUTHORS Lauersen KJ, Mussgnug JH, Kruse O.
TITLE Direct Submission
JOURNAL Submitted (18-MAY-2016) Algae Biotechnology and Bioenergy,
University of Bielefeld Center for Biotechnology, Universitaetsstr
27, Bielefeld, NRW 33615, Germany
REFERENCE 4 (bases 1 to 6831)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6831)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(25-JUN-2014) Algae Biotechnology and Bioenergy, University of
Bielefeld Center for Biotechnology, Universitaetsstr 27, Bielefeld,
NRW 33615, Germany"
SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(18-MAY-2016) Algae Biotechnology and Bioenergy, University of
Bielefeld Center for Biotechnology, Universitaetsstr 27, Bielefeld,
NRW 33615, Germany"
SGRef: number: 4; type: "Journal Article"
On May 18, 2016 this sequence version replaced KM061067.1.
FEATURES Location/Qualifiers
source 1..6831
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 489..505
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 515..533
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1247..1258
/label="Factor Xa site"
/note="Factor Xa recognition and cleavage site"
CDS 2269..2280
/label="Factor Xa site"
/note="Factor Xa recognition and cleavage site"
CDS 2293..2316
/label="Strep-Tag II"
/note="peptide that binds Strep-Tactin(R), an engineered
form of streptavidin"
CDS 3266..4261
/gene="hyg"
/label="Hygromycin-B 7''-O-kinase"
/note="Hygromycin-B 7''-O-kinase from Streptomyces
hygroscopicus. Accession#: P09979"
promoter complement(4531..4549)
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4570..4586)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4594..4610)
/label="lac operator"
/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(4618..4648)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(4663..4684)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4972..5560)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5734..6591)
/label="AmpR"
/note="beta-lactamase"
promoter complement(6592..6696)
/label="AmpR promoter"
rep_origin complement(join(6723..6831,1..347))
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"