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。
pCS2+ contains a powerful enhancer/promoter (simian CMV ie94). In the 5' untranslated region of the mRNA transcribed from the sCMV promoter, there is an SP6 promoter for in vitro RNA synthesis. It also has a T7 promoter in the opposite direction between the multiple cloning site and the SV40 polyadenylation site for probe synthesis. Its backbone is from pBluescriptIKS+.
pCS2+ can have high-level transient expression in multiple cell types and for in vitro transcription and translation. It is a great choice when researchers need a vector for gene expression studies in various organisms such as mammalian cells, Xenopus cells, avian cells, and zebrafish cells.
- 载体名称:
- pCS2+
- 载体抗性:
- Ampicillin
- 载体长度:
- 4092 bp
- 载体类型:
- I.M.A.G.E. Consortium Plasmids
- 复制子:
- ori
- 载体来源:
- I.M.A.G.E. Consortium
- 拷贝数:
- High copy number
- 启动子:
- sCMV
- 3'引物:
- M13 rev
- 感受态:
- JM108
- 培养温度:
- 37℃
pCS2+ 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。
pCS2+ 载体序列
LOCUS Exported 4092 bp DNA circular SYN 03-SEP-2024
DEFINITION Vector that allows high-level transient expression in vertebrate
cells as well as in vitro transcription/translation.
ACCESSION .
VERSION .
KEYWORDS pCS2+
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4092)
AUTHORS I.M.A.G.E. Consortium
TITLE Direct Submission
REFERENCE 2 (bases 1 to 4092)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 4092)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4092
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 35..53
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
misc_feature 117..144
/label=stop codons
/note="stop codons"
/note="stop codons in all three reading frames"
polyA_signal complement(144..278)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(391..409)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(430..446)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(454..470)
/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(478..508)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(523..544)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(832..1420)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1594..2451)
/label=AmpR
/note="beta-lactamase"
promoter complement(2452..2556)
/label=AmpR promoter
rep_origin complement(2582..3037)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3111..4092
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"