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:
- pSKA563
- 载体抗性:
- Ampicillin
- 载体长度:
- 8589 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Aoki SK, Lillacci G, Gupta A, Baumschlager A
pSKA563 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。
pSKA563 载体序列
LOCUS V015319 8589 bp DNA circular SYN 26-JUN-2019
DEFINITION Exported.
ACCESSION V015319
VERSION V015319
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 8589)
AUTHORS Aoki SK, Lillacci G, Gupta A, Baumschlager A, Schweingruber D,
Khammash M.
TITLE A universal biomolecular integral feedback controller for robust
perfect adaptation
JOURNAL Nature 570, 533-537 (2019)
PUBMED 31217585
REFERENCE 2 (bases 1 to 8589)
AUTHORS Aoki SK, Lillacci G.
TITLE Direct Submission
JOURNAL Submitted (09-APR-2019) Biosystems Science and Engineering, ETH
Zurich, Mattenstrasse 26, Basel 4058, Switzerland
REFERENCE 3 (bases 1 to 8589)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature 570,
533-537 (2019)"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-APR-2019) Biosystems Science and Engineering, ETH Zurich,
Mattenstrasse 26, Basel 4058, Switzerland"
FEATURES Location/Qualifiers
source 1..8589
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 26..47
/label="UP element"
/note="UP element"
/regulatory_class="enhancer"
regulatory 50..85
/label="PsigW"
/note="PsigW"
/regulatory_class="promoter"
regulatory 112..118
/label="sRBS"
/note="sRBS"
/regulatory_class="ribosome_binding_site"
CDS 128..169
/label="V5 tag"
/note="epitope tag from simian virus 5"
CDS 176..1048
/label="araC"
/note="L-arabinose regulatory protein"
regulatory 1139..1145
/label="sRBS"
/note="sRBS"
/regulatory_class="ribosome_binding_site"
CDS 1155..1178
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 1191..1901
/label="superfolder GFP"
/note="GFP variant that folds robustly even when fused to
poorly folded proteins (Pedelacq et al., 2006)"
terminator 2206..2292
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2384..2411
/label="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
regulatory 2494..2515
/label="UP element"
/note="UP element"
/regulatory_class="enhancer"
regulatory 2518..2553
/label="PsigW"
/note="PsigW"
/regulatory_class="promoter"
regulatory 2580..2586
/label="sRBS"
/note="sRBS"
/regulatory_class="ribosome_binding_site"
CDS 2596..2637
/label="V5 tag"
/note="epitope tag from simian virus 5"
CDS 2644..3516
/label="araC"
/note="L-arabinose regulatory protein"
regulatory 3607..3613
/label="sRBS"
/note="sRBS"
/regulatory_class="ribosome_binding_site"
CDS 3623..3646
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 3659..4369
/label="superfolder GFP"
/note="GFP variant that folds robustly even when fused to
poorly folded proteins (Pedelacq et al., 2006)"
regulatory 4472..4897
/label="rrnB T1T2 terminator"
/note="rrnB T1T2 terminator"
/regulatory_class="terminator"
terminator complement(4973..5020)
/label="T7 terminator"
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(5024..5773)
/gene="luxR"
/label="Transcriptional activator protein LuxR"
/note="Transcriptional activator protein LuxR from
Aliivibrio fischeri. Accession#: P12746"
regulatory complement(5802..5855)
/label="Pconstitutive"
/note="Pconstitutive"
/regulatory_class="promoter"
regulatory 5906..5970
/label="Plux"
/note="Plux"
/regulatory_class="promoter"
regulatory 5906..5925
/label="LuxR binding site"
/note="LuxR binding site"
/regulatory_class="other"
regulatory 5971..5996
/label="RBS5000"
/note="RBS5000"
/regulatory_class="ribosome_binding_site"
CDS 5997..6557
/gene="sigW"
/label="ECF RNA polymerase sigma factor SigW"
/note="ECF RNA polymerase sigma factor SigW from Bacillus
subtilis (strain 168). Accession#: Q45585"
terminator complement(6575..6618)
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin complement(6760..7348)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator complement(7436..7530)
/label="lambda t0 terminator"
/note="transcription terminator from phage lambda"
CDS complement(7563..8420)
/label="AmpR"
/note="beta-lactamase"
promoter complement(8421..8525)
/label="AmpR promoter"