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:
- pCE-hUL
- 载体抗性:
- Ampicillin
- 载体长度:
- 11232 bp
- 载体类型:
- Mammalian Expression
- 复制子:
- ori
- 拷贝数:
- High Copy
- 启动子:
- CAG
- 克隆方法:
- Restriction Enzyme
- 5'引物:
- pCAG-F
- 3'引物:
- TTAGCCAGAAGTCAGATGCTC
- 感受态:
- DH5a
pCE-hUL 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。
pCE-hUL 载体序列
LOCUS V000584 11232 bp DNA circular SYN 10-JUN-2021
DEFINITION Exported.
ACCESSION V000584
VERSION V000584
KEYWORDS pCE-hUL
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11232)
AUTHORS Okita K, Yamakawa T, Matsumura Y, Sato Y, Amano N, Watanabe A,
Goshima N, Yamanaka S
TITLE An Efficient Non-viral Method to Generate Integration-Free Human iPS
Cells from Cord Blood and Peripheral Blood Cells.
JOURNAL Stem Cells. 2012 Nov 29. doi: 10.1002/stem.1293.
PUBMED 23193063
REFERENCE 2 (bases 1 to 11232)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 11232)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells.
2012 Nov 29. doi: 10.1002/stem.1293."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11232
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(83..102)
/label="Bglob-pA-R"
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 148..203
/label="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(202..221)
/label="rbglobpA-R"
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(564..580)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(588..604)
/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(612..642)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(657..678)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
CDS 1157..3079
/codon_start=1
/product="Epstein-Barr nuclear antigen 1, also known as
EBNA-1"
/label="EBNA1"
/note="crucial for latent viral infection, and for episomal
amplification of vectors containing the oriP origin (Okita
et al., 2013)"
/translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR
GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG
AGGGAGAGGGAGGAGGAGGAGAGGGAGXGGGAGGAGGAGAGGGAGXGGGAGGAGAGGGA
GGAGGAGAGGGAGAGGGAGGAGAGGGAGGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG
GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGAGGAGGXGAGGAGGAGAGG
GAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRGG
SGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPPP
GRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGRR
KKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSLY
NLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLKD
AIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDEG
EEGQE"
rep_origin 3381..5170
/label="oriP"
/note="Epstein-Barr virus oriP replication origin (Yates et
al., 2000)"
primer_bind complement(5711..5728)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(5882..6470)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6644..7501)
/label="AmpR"
/note="beta-lactamase"
promoter complement(7502..7606)
/label="AmpR promoter"
enhancer 7637..8016
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 8018..8295
/label="chicken beta-actin promoter"
intron 8296..9312
/label="chimeric intron"
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 9320..9339
/label="pCAG-F"
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 9377..10468
/gene="MYCL"
/label="Protein L-Myc"
/note="Protein L-Myc from Homo sapiens. Accession#: P12524"
CDS 10493..10558
/label="F2A"
/note="2A peptide from foot-and-mouth disease virus
polyprotein"
CDS 10583..11209
/label="hLIN28A"
/note="Human lin-28 homolog A gene. Encodes a LIN-28 family
RNA-binding protein that acts as a posttranscriptional
regulator of genes involved in development, self-renewal of
embryonic stem cells and metabolism. Overexpressed in human
embryonic stem cells, primary human tumors and human cancer
cell lines."