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。
For e.coli protein expression.
- 载体名称:
- pCWori
- 载体抗性:
- Ampicillin
- 载体长度:
- 5438 bp
- 载体类型:
- Prokaryotic Expression Vectors
- 复制子:
- ori
- 启动子:
- tac/lac
- 感受态:
- DH10B
- 培养温度:
- 37℃
pCWori 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。
pCWori 载体序列
LOCUS Exported 5438 bp DNA circular SYN 31-AUG-2024
DEFINITION Exported.
ACCESSION V001682
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5438)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5438)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5438)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5438
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 644..649
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 1126..1131
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 21..407
/gene="cheY"
/label=Chemotaxis protein CheY
/note="Chemotaxis protein CheY from Escherichia coli
(strain K12). Accession#: P0AE67"
primer_bind complement(465..481)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 668..1123
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1156..1260
/label=AmpR promoter
CDS 1261..2118
/label=AmpR
/note="beta-lactamase"
rep_origin 2292..2880
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(3066..3207)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(3616..3789)
/label=lacZ-alpha
/note="LacZ-alpha fragment of beta-galactosidase"
protein_bind complement(3809..3825)
/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(3833..3863)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3878..3899)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(3915..4994)
/label=lacI
/note="lac repressor"
promoter complement(4995..5072)
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
protein_bind 5124..5145
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5160..5190
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
protein_bind 5198..5214
/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 5269..5297
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 5305..5321
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="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)."
misc_feature 5364..5392
/label=tac promoter
/note="tac promoter"
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 5400..5416
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="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)."