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:
- AbVec-hIgG1
- 载体抗性:
- Ampicillin
- 载体长度:
- 5746 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Di Niro R, Mesin L, Raki M, Zheng NY, Lund-Johansen F, Lundin KE, Charpilienne A, Poncet D, Wilson PC, Sollid LM.
- 启动子:
- SV40
AbVec-hIgG1 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。
AbVec-hIgG1 载体序列
LOCUS 40924_190 5746 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector AbVec-hIgG1, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5746) AUTHORS Di Niro R, Mesin L, Raki M, Zheng NY, Lund-Johansen F, Lundin KE, Charpilienne A, Poncet D, Wilson PC, Sollid LM. TITLE Rapid Generation of Rotavirus-Specific Human Monoclonal Antibodies from Small-Intestinal Mucosa JOURNAL J. Immunol. 185 (9), 5377-5383 (2010) PUBMED 20935207 REFERENCE 2 (bases 1 to 5746) AUTHORS Wilson PC. TITLE Antibody variable gene expression vector for human IgG1 heavy chain JOURNAL Unpublished REFERENCE 3 (bases 1 to 5746) AUTHORS Wilson PC. TITLE Direct Submission JOURNAL Submitted (18-NOV-2008) Medicine/Rheumatology, The University of Chicago, BSLC/JFK Bldg. 924 East 57th St., Rm 414, Chicago, IL 60637, USA REFERENCE 4 (bases 1 to 5746) TITLE Direct Submission REFERENCE 5 (bases 1 to 5746) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Immunol."; date: "2010"; volume: "185"; issue: "9"; pages: "5377-5383" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (18-NOV-2008) Medicine/Rheumatology, The University of Chicago, BSLC/JFK Bldg. 924 East 57th St., Rm 414, Chicago, IL 60637, USA" COMMENT SGRef: number: 4; type: "Journal Article" FEATURES Location/Qualifiers source 1..5746 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 14..393 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 394..597 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 828..846 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" CDS 929..985 /label=IgH signal sequence /note="signal sequence from the mouse immunoglobulin heavy chain variable region" CDS 995..1981 /label=hlgG1-CH /note="Human IgG1 heavy chain constant region" regulatory 1991..2000 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" polyA_signal 2004..2138 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter 2211..2568 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2588..2604) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 2817..3272 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3604..3708 /label=AmpR promoter CDS 3709..4566 /label=AmpR /note="beta-lactamase" rep_origin 4740..5328 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 5616..5637 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 5652..5682 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5690..5706 /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)." primer_bind 5714..5730 /label=M13 rev /note="common sequencing primer, one of multiple similar variants"