Note: Ubiquitous Chromatin Opening Element (UCOE) technology significantly enhances gene expression in stably transfected mammalian cells by modulating chromatin structure. Derived from small DNA elements near housekeeping genes, UCOEs create a transcriptionally active open chromatin environment around the integrated transgene. This prevents transgene silencing and enables consistent, stable, and high-level gene expression regardless of the transgene's chromosomal integration site.
基本信息
- 载体名称:
- UCOE Mu-H
- 载体抗性:
- Ampicillin
- 载体长度:
- 8182 bp
- 载体类型:
- Protein expression
- 复制子:
- ori
- 宿主:
- Mammalian cells
- 筛选标记:
- Hyg
- 启动子:
- CMV
- 感受态:
- DH10B
- 培养温度:
- 37℃
产品信息
质粒编号:V013488
质粒名称:UCOE Mu-H
规格:5 μg (冻干粉)
下载资源
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。
确保质粒的关键元件正确，但是我们并不能保证实验效果。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。
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。
UCOE Mu-H 质粒 (编号: V013488)序列
LOCUS Exported 8182 bp DNA circular SYN 03-DEC-2025
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8182)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 8182)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8182
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 31..47
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 57..75
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 158..174
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 249..3296
/label=UCOE
/note="The Rps3 chromatin opening element (UCOE) is a
fragment of DNA from the mouse Rps3 gene that is used in
biotechnology and gene therapy to maintain stable and
high-level gene expression. It functions as a ubiquitous
chromatin opening element that prevents epigenetic
silencing by keeping the DNA in an open, transcriptionally
active state. "
enhancer 3383..3686
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3687..3890
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
polyA_signal 4142..4263
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 4387..4890
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 4925..5947
/codon_start=1
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
E"
promoter complement(6017..6035)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6056..6072)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(6080..6096)
/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(6104..6134)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6149..6170)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6458..7046)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7220..8077)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8078..8182)
/label=AmpR promoter