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:
- pETcoco-2
- 载体抗性:
- Ampicillin
- 载体长度:
- 12417 bp
- 载体类型:
- pET & Duet Vectors (Novagen)
- 复制子:
- ori2
- 载体来源:
- Novagen (EMD Millipore)
- 拷贝数:
- Low copy number
- 启动子:
- araBAD
pETcoco-2 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。
pETcoco-2 载体序列
LOCUS pETcoco-2. 12417 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial vector with an ampicillin resistance marker that allows
single-copy replication, on-demand amplification, and tightly
regulated expression.
ACCESSION .
VERSION .
KEYWORDS pETcoco-2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12417)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 12417)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT The copy number can be increased to 20-50 copies per cell by adding
arabinose to the growth medium.
FEATURES Location/Qualifiers
source 1..12417
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 3..27
/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)."
RBS 42..64
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 72..74
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 75..92
/label=6xHis
/note="6xHis affinity tag"
CDS 102..146
/label=S-Tag
/note="affinity and epitope tag derived from pancreatic
ribonuclease A"
CDS 162..176
/label=enterokinase site
/note="enterokinase recognition and cleavage site"
CDS 249..281
/label=HSV tag
/note="HSV (herpes simplex virus) epitope tag"
CDS 326..343
/label=6xHis
/note="6xHis affinity tag"
terminator 410..457
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(834..1709)
/label=araC
/note="L-arabinose regulatory protein"
promoter 1736..2020
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
CDS 2080..3225
/label=trfA
/note="trans-acting replication protein that binds to and
activates oriV"
CDS complement(3449..4306)
/label=AmpR
/note="beta-lactamase"
promoter complement(4307..4398)
/label=AmpR promoter
rep_origin complement(4965..5677)
/direction=LEFT
/label=oriV
/note="incP origin of replication"
rep_origin 5756..5975
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
CDS 6066..6818
/label=repE
/note="replication initiation protein for the bacterial F
plasmid"
misc_feature 6824..7074
/label=incC
/note="incompatibility region of the bacterial F plasmid"
CDS 7400..8572
/label=sopA
/note="partitioning protein for the bacterial F plasmid"
CDS 8575..9543
/label=sopB
/note="partitioning protein for the bacterial F plasmid"
misc_feature 9619..10092
/gene="sopC"
/label=sopC
/note="sopC"
/note="centromere-like partitioning region of the bacterial
F plasmid"
misc_feature complement(10352..10750)
/label=cos
/note="lambda cos site; allows packaging into phage lambda
particles"
protein_bind complement(10768..10801)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
protein_bind complement(10894..10915)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(10931..12010)
/label=lacI
/note="lac repressor"
promoter complement(12011..12088)
/label=lacI promoter