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:
- pDUAL-HFF21
- 载体抗性:
- Ampicillin
- 载体长度:
- 8770 bp
- 载体类型:
- S. pombe expression vector
- 复制子:
- ori
- 载体来源:
- Matsuyama A, Shirai A, Yashiroda Y, Kamata A, Horinouchi S, Yoshida M.
pDUAL-HFF21 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。
pDUAL-HFF21 载体序列
LOCUS 40924_15920 8770 bp DNA circular SYN 17-DEC-2018
DEFINITION S. pombe expression vector pDUAL-HFF21 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8770)
AUTHORS Matsuyama A, Shirai A, Yashiroda Y, Kamata A, Horinouchi S, Yoshida
M.
TITLE pDUAL, a multipurpose, multicopy vector capable of chromosomal
integration in fission yeast
JOURNAL Yeast 21 (15), 1289-1305 (2004)
PUBMED 15546162
REFERENCE 2 (bases 1 to 8770)
AUTHORS Matsuyama A, Shirai A, Yoshida M.
TITLE A series of promoters for constitutive expression of heterologous
genes in fission yeast
JOURNAL Yeast 25 (5), 371-376 (2008)
PUBMED 18437702
REFERENCE 3 (bases 1 to 8770)
AUTHORS Matsuyama A, Shirai A, Yoshida M.
TITLE Direct Submission
JOURNAL Submitted (15-OCT-2007) Contact:Akihisa Matsuyama RIKEN, Chemical
Genetics Laboratory; Hirosawa 2-1, Wako, Saitama 351-0198, Japan URL
:http://cgl.riken.go.jp/
REFERENCE 4 (bases 1 to 8770)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 8770)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Yeast";
date: "2004"; volume: "21"; issue: "15"; pages: "1289-1305"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Yeast";
date: "2008"; volume: "25"; issue: "5"; pages: "371-376"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(15-OCT-2007) Contact:Akihisa Matsuyama RIKEN, Chemical Genetics
Laboratory; Hirosawa 2-1, Wako, Saitama 351-0198, Japan URL
:http://cgl.riken.go.jp/"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT Fission yeast expression vector.
FEATURES Location/Qualifiers
source 1..8770
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 21..208
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
regulatory 244..954
/pseudo
/gene="leu1"
/regulatory_class="promoter"
regulatory complement(1483..1871)
/gene="ura4"
/regulatory_class="terminator"
gene complement(1483..1871)
/gene="ura4"
/label=ura4
CDS complement(1875..2666)
/label=S. pombe ura4
/note="orotidine 5'-phosphate decarboxylase, required for
uracil biosynthesis"
regulatory complement(2667..3190)
/gene="ura4"
/regulatory_class="promoter"
gene complement(2667..3190)
/gene="ura4"
/label=ura4
rep_origin complement(3275..4058)
/direction=LEFT
/label=ars1
/note="Schizosaccharomyces pombe autonomously replicating
sequence ars1"
primer_bind complement(4781..4797)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 5010..5465
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5747..5851
/label=AmpR promoter
CDS 5852..6709
/label=AmpR
/note="beta-lactamase"
rep_origin 6883..7471
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 7759..7780
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7795..7825
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7833..7849
/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)."
primer_bind 7857..7873
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory 7898..8636
/gene="tub1"
/regulatory_class="promoter"
misc_feature 7898..8636
/gene="tub1"
/label=derived from Schizosaccharomyces pombe
/note="derived from Schizosaccharomyces pombe"
gene 7898..8636
/gene="tub1"
/label=tub1
CDS 8665..8682
/label=6xHis
/note="6xHis affinity tag"
CDS 8686..8709
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 8716..8739
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"