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:
- YCplac22 YCp-She3-TG
- 载体抗性:
- Ampicillin
- 载体长度:
- 9485 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Belmont BJ, Niles JC.
- 启动子:
- TRP1
YCplac22 YCp-She3-TG 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。
YCplac22 YCp-She3-TG 载体序列
LOCUS V001501 9485 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V001501
VERSION V001501
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9485)
AUTHORS Belmont BJ, Niles JC.
TITLE Inducible Control of Subcellular RNA Localization Using a Synthetic
Protein-RNA Aptamer Interaction
JOURNAL PLoS ONE 7 (10), E46868 (2012)
PUBMED 23056498
REFERENCE 2 (bases 1 to 9485)
AUTHORS Belmont BJ, Niles JC.
TITLE Direct Submission
JOURNAL Submitted (12-SEP-2012) Biological Engineering, Massachusetts
Institute of Technology, 77 Massachusetts Ave, Cambridge, MA 02139,
USA
REFERENCE 3 (bases 1 to 9485)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9485)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
date: "2012"; volume: "7"; issue: "10"; pages: "E46868"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-SEP-2012) Biological Engineering, Massachusetts Institute of
Technology, 77 Massachusetts Ave, Cambridge, MA 02139, USA"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9485
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 106..127
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 142..172
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 180..196
/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 204..220
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
CDS 1239..2513
/gene="SHE3"
/label="SWI5-dependent HO expression protein 3"
/note="SWI5-dependent HO expression protein 3 from
Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
Accession#: P38272"
CDS 2532..3149
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label="TetR"
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
CDS complement(3154..3180)
/label="9xHis"
/note="9xHis affinity tag"
CDS 3180..3893
/label="EGFP"
/note="enhanced GFP"
CDS 4320..4340
/label="7xHis"
/note="6xHis affinity tag"
primer_bind complement(4921..4937)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(5696..6858)
/label="CEN/ARS"
/note="S. cerevisiae CEN4 centromere fused to the
autonomously replicating sequence ARS1/ARS416"
promoter complement(7372..7473)
/label="TRP1 promoter"
promoter 7579..7683
/label="AmpR promoter"
CDS 7684..8541
/label="AmpR"
/note="beta-lactamase"
rep_origin 8715..9303
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"