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:
- pRLYC2
- 载体抗性:
- Ampicillin
- 载体长度:
- 5627 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Redder P, Linder P.
- 启动子:
- lac
pRLYC2 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。
pRLYC2 载体序列
LOCUS V003691 5627 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V003691
VERSION V003691
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 5627)
AUTHORS Redder P, Linder P.
TITLE New Range of Vectors with a Stringent 5-Fluoroorotic Acid-Based
Counterselection System for Generating Mutants by Allelic
Replacement in Staphylococcus aureus
JOURNAL Appl. Environ. Microbiol. 78 (11), 3846-3854 (2012)
PUBMED 22447609
REFERENCE 2 (bases 1 to 5627)
AUTHORS Redder P, Linder P.
TITLE Direct Submission
JOURNAL Submitted (12-MAR-2012) Department of Microbiology and Molecular
Medicine, University of Geneva, Faculty of Medicine, Rue
Michel-Servet 1, Geneva 1203, Switzerland
REFERENCE 3 (bases 1 to 5627)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5627)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Assembly Method :: BioEdit, CAP v. 7.0.9.0
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2012"; volume: "78"; issue: "11";
pages: "3846-3854"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-MAR-2012) Department of Microbiology and Molecular Medicine,
University of Geneva, Faculty of Medicine, Rue Michel-Servet 1,
Geneva 1203, Switzerland"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5627
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 396..452
/label="MCS"
/note="pUC18/19 multiple cloning site"
primer_bind complement(465..481)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(489..505)
/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(513..543)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(558..579)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(717..1364)
/gene="pyrE"
/label="Orotate phosphoribosyltransferase"
/note="Orotate phosphoribosyltransferase from Bacillus
subtilis (strain 168). Accession#: P25972"
CDS complement(1346..2062)
/gene="pyrF"
/label="Orotidine 5'-phosphate decarboxylase"
/note="Orotidine 5'-phosphate decarboxylase from Bacillus
subtilis (strain 168). Accession#: P25971"
CDS complement(2424..3071)
/gene="cat"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
rep_origin 3457..3667
/note="derived from pT181; allows replication in
Staphylococcus aureus if the RepC protein is present;
ORI181"
rep_origin complement(3808..4396)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4570..5427)
/label="AmpR"
/note="beta-lactamase"
promoter complement(5428..5532)
/label="AmpR promoter"