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:
- pCAGGS-E-hABIN-2
- 载体抗性:
- Ampicillin
- 载体长度:
- 6557 bp
- 载体类型:
- Mammalian expression vector
- 复制子:
- ori
- 载体来源:
- De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R.
- 启动子:
- CAG
pCAGGS-E-hABIN-2 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。
pCAGGS-E-hABIN-2 载体序列
LOCUS V008879 6557 bp DNA circular SYN 17-DEC-2018
DEFINITION Exported.
ACCESSION V008879
VERSION V008879
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6557)
AUTHORS De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman
Fonseca M, Vanhoucke M, Beyaert R.
TITLE BCCM/LMBP Plasmid collection
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6557)
AUTHORS De Schamphelaire W.
TITLE Direct Submission
JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent,
Technologiepark 927, 9052, BELGIUM
REFERENCE 3 (bases 1 to 6557)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6557)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927,
9052, BELGIUM"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6557
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 13..51
/label="E-tag"
/note="E-tag"
CDS 64..1350
/gene="TNIP2"
/label="TNFAIP3-interacting protein 2"
/note="TNFAIP3-interacting protein 2 from Homo sapiens.
Accession#: Q8NFZ5"
polyA_signal 1914..1969
/label="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2330..2346)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(2354..2370)
/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(2378..2408)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(2422..2443)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2501..2697
/label="SV40 promoter"
/note="SV40 early promoter"
polyA_signal 2703..2837
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(3075..3663)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(3837..4694)
/label="AmpR"
/note="beta-lactamase"
promoter complement(4695..4799)
/label="AmpR promoter"
enhancer 4830..5209
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
regulatory 4830..5207
/label="hCMV-IE enhancer"
/note="hCMV-IE enhancer"
/regulatory_class="promoter"
promoter 5211..5488
/label="chicken beta-actin promoter"
intron 5489..6506
/label="chimeric intron"
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"