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A, Meert J, Verhelst K, Roggeman Fonseca M, Vanhoucke M, Beyaert R. TITLE BCCM/LMBP Plasmid collection JOURNAL Unpublished REFERENCE 2 (bases 1 to 6557) AUTHORS De Schamphelaire W. TITLE Direct Submission JOURNAL Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark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 TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 9052, BELGIUM" SGRef: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"