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。
The E06 antibody is a murine monoclonal IgM natural antibody that binds to oxidized phospholipid and reacts with oxidized LDL, oxidized HDL and protein covalently modified by oxidized phospholipid. This antibody has been used to probe atherosclerotic plaque in mice, zebrafish, rabbits, monkeys and humans. It also binds to apoptotic cells, but not viable cells, and can be used in a variety of applications, including ELISA, immunohistochemistry and Western blot analysis, as well as for in vivo imaging with MRI techniques.
- 载体名称:
- pSecTag2A-E06-scFv
- 载体抗性:
- Ampicillin
- 载体长度:
- 5846 bp
- 载体类型:
- Mammalian Expression Vectors
- 复制子:
- ori
- 载体来源:
- Dr. Joseph Witztum
- 筛选标记:
- Zeocin
- 拷贝数:
- High copy number
- 启动子:
- CMV
- 克隆方法:
- Enzyme Cut
- 5'引物:
- T7 promoter
- 3'引物:
- BGH Reverse primer
- 载体标签:
- Myc,6*His
- 感受态:
- JM108
- 培养温度:
- 37℃
pSecTag2A-E06-scFv 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。
pSecTag2A-E06-scFv 载体序列
LOCUS Exported 5846 bp DNA circular SYN 02-SEP-2024
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS pSecTag2A-E06-scFv
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5846)
AUTHORS Triple Threat
TITLE Direct Submission
REFERENCE 2 (bases 1 to 5846)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5846)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5846
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
sig_peptide 905..967
/label=Ig-kappa leader
/note="leader sequence from mouse immunoglobulin kappa
light
chain"
CDS 968..1342
/codon_start=1
/label=E06 VL
/note="E06 VL"
/translation="AAQPARRAVRSLDIVMTQSPSSLSVSAGKKVTISCTASESLYSSK
HKVHYLAWYQKKPEQSPKLLIYGASNRYIGVPDRFTGSGSGTDFTLTISSVQVEDLTHY
YCAQFYSYPLTFGAGTKLEIK"
CDS 1343..1387
/codon_start=1
/label=G4S x 3
/note="G4S x 3"
/translation="GGGGSGGGGSGGGGS"
CDS 1388..1768
/codon_start=1
/label=E06 VH
/note="E06 VH"
/translation="EVKLVESGGGLVQPGGSLRLSCATSGFTFSDFYMEWVRQAPGKRL
EWIAASRNKANDYTTEYADSVKGRFIVSRDTSQSILYLQMNALRAEDTAIYYCARDYYG
SSYWYFDVWGAGTTVTVSSRGGP"
CDS 1769..1798
/codon_start=1
/product="Myc (human c-Myc oncogene) epitope tag"
/label=Myc
/note="Myc"
/translation="EQKLISEEDL"
CDS 1814..1831
/label=6xHis
/note="6xHis affinity tag"
polyA_signal 1860..2084
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2130..2558
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2572..2901
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 2949..2996
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 3015..3386
/label=BleoR
/note="antibiotic-binding protein"
polyA_signal 3519..3652
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3689..3705)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3713..3729)
/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(3737..3767)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3782..3803)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4091..4679)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4853..5710)
/label=AmpR
/note="beta-lactamase"
promoter complement(5711..5815)
/label=AmpR promoter