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:
- pAS2hA20R562A
- 载体抗性:
- Ampicillin
- 载体长度:
- 10805 bp
- 载体类型:
- Yeast expression vector
- 复制子:
- ori
- 载体来源:
- De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R.
- 启动子:
- ADH1(medium)
pAS2hA20R562A 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。
pAS2hA20R562A 载体序列
LOCUS V009428 10805 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V009428
VERSION V009428
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10805)
AUTHORS De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman
Fonseca M, Vanhoucke M, Beyaert R.
TITLE BCCM/LMBP Plasmid collection
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 10805)
AUTHORS De Schamphelaire W.
TITLE Direct Submission
JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent,
Technologiepark 927, 9052, BELGIUM
REFERENCE 3 (bases 1 to 10805)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10805)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
9052, BELGIUM"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10805
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind complement(965..1012)
/label="FRT"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter 1603..1883
/label="TRP1 promoter"
CDS 1884..2555
/label="TRP1"
/note="phosphoribosylanthranilate isomerase, required for
tryptophan biosynthesis"
rep_origin complement(2661..3116)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 3261..3277
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
terminator complement(3314..3561)
/label="CYC1 terminator"
/note="transcription terminator for CYC1"
CDS complement(3858..4304)
/gene="RPL28"
/label="Large ribosomal subunit protein uL15"
/note="Large ribosomal subunit protein uL15 from
Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
Accession#: P02406"
promoter 4769..5473
/label="ADH1 promoter"
/note="promoter for alcohol dehydrogenase 1"
CDS 5501..5941
/label="GAL4 DNA binding domain"
/note="DNA binding domain of the GAL4 transcriptional
activator"
CDS 5957..5983
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 5999..8368
/gene="TNFAIP3"
/label="Tumor necrosis factor alpha-induced protein 3"
/note="Tumor necrosis factor alpha-induced protein 3 from
Homo sapiens. Accession#: P21580"
terminator 8443..8630
/label="ADH1 terminator"
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(8654..8670)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8678..8694
/label="lac operator"
/bound_moiety="lac repressor encoded by lacI"
/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(8702..8732)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(8747..8768)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(9056..9644)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9818..10675)
/label="AmpR"
/note="beta-lactamase"
promoter complement(10676..10780)
/label="AmpR promoter"