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:
- pDIS-URA3-Clox
- 载体抗性:
- Ampicillin
- 载体长度:
- 9539 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F
- 启动子:
- TRP1
pDIS-URA3-Clox 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。
pDIS-URA3-Clox 载体序列
LOCUS V015620 9539 bp DNA circular SYN 17-JUL-2019
DEFINITION Exported.
ACCESSION V015620
VERSION V015620
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9539)
AUTHORS Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F,
Correa-Bordes J, Vazquez de Aldana CR.
TITLE A new toolkit for gene tagging in Candida albicans containing
recyclable markers
JOURNAL PLoS ONE (2019) In press
REFERENCE 2 (bases 1 to 9539)
AUTHORS Duenas-Santero E, Santos-Almeida A, Rojo-Dominguez P, del Rey F,
Correa-Bordes J, Vazquez de Aldana CR.
TITLE Direct Submission
JOURNAL Submitted (18-MAR-2019) Instituto de Biologia Funcional y Genomica
(IBFG), Consejo Superior de Investigaciones Cientificas, Zacarias
Gonzalez 2, Salamanca, Salamanca 37007, Spain
REFERENCE 3 (bases 1 to 9539)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Assembly Method :: Lasergene v. 13
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE
(2019) In press"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(18-MAR-2019) Instituto de Biologia Funcional y Genomica (IBFG),
Consejo Superior de Investigaciones Cientificas, Zacarias Gonzalez
2, Salamanca, Salamanca 37007, Spain"
FEATURES Location/Qualifiers
source 1..9539
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(166..465)
/label="NEUT5L 3'"
/note="NEUT5L 3'"
primer_bind 736..752
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 759..777
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 817..833
/label="SK primer"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 847..880
/label="loxP"
/note="loxP"
CDS 1310..2119
/gene="URA3"
/label="Orotidine 5'-phosphate decarboxylase"
/note="Orotidine 5'-phosphate decarboxylase from Candida
albicans (strain SC5314 / ATCC MYA-2876). Accession#:
P13649"
regulatory 2264..3595
/note="CaMET3 promoter; inducible upon growth without
methionine repressible via addition of methionine and
cysteine to the medium"
/regulatory_class="promoter"
CDS join(3623..4026,4191..4818)
/codon_start=1
/transl_table=11
/product="Cre"
/label="Cre"
/note="Cre recombinase"
/protein_id="QDK59802.1"
/translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
DGD"
intron 4027..4190
/note="modified TUB2 intron sequence"
3'UTR 4831..4961
/label="Saccharomyces cerevisiae ADH1 3'UTR region"
/note="Saccharomyces cerevisiae ADH1 3'UTR region"
regulatory 4962..5021
/note="Transcriptional termination from Saccharomyces
cerevisiae CYC1 gene"
/regulatory_class="terminator"
protein_bind complement(5034..5067)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
primer_bind complement(5147..5163)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(5174..5423)
/label="NEUT5L 5'"
/note="NEUT5L 5'"
promoter complement(5438..5468)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(5483..5504)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5792..6380)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6554..7411)
/label="AmpR"
/note="beta-lactamase"
promoter complement(7412..7516)
/label="AmpR promoter"
misc_feature 7553..8056
/label="CEN/ARS"
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 8312..8592
/label="TRP1 promoter"
CDS 8593..9264
/label="TRP1"
/note="phosphoribosylanthranilate isomerase, required for
tryptophan biosynthesis"