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:
- pHsCXW
- 载体抗性:
- Ampicillin
- 载体长度:
- 7207 bp
- 载体类型:
- Lentiviral transfer vector
- 复制子:
- ori
- 载体来源:
- Leander Johansen J, Dago L, Tornoe J, Rosenblad C, Kusk P.
- 启动子:
- CMV
pHsCXW 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。
pHsCXW 载体序列
LOCUS V005488 7207 bp DNA circular SYN 18-DEC-2018 DEFINITION Exported. ACCESSION V005488 VERSION V005488 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 7207) AUTHORS Leander Johansen J, Dago L, Tornoe J, Rosenblad C, Kusk P. TITLE A new versatile and compact lentiviral vector JOURNAL Mol. Biotechnol. 29 (1), 47-56 (2005) PUBMED 15668519 REFERENCE 2 (bases 1 to 7207) AUTHORS Johansen J, Tornoe J, Rosenblad C, Dago L, Kusk P. TITLE Improved lentiviral transfer vector JOURNAL Unpublished REFERENCE 3 (bases 1 to 7207) AUTHORS Kusk P, Johansen J. TITLE Direct Submission JOURNAL Submitted (19-NOV-2003) NsGene A/S, Baltorpvej 154, Ballerup DK-2750, Denmark REFERENCE 4 (bases 1 to 7207) TITLE Direct Submission REFERENCE 5 (bases 1 to 7207) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Biotechnol."; date: "2005"; volume: "29"; issue: "1"; pages: "47-56" SGRef: number: 2; type: "Journal Article"; journalName: "Unpublished" SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (19-NOV-2003) NsGene A/S, Baltorpvej 154, Ballerup DK-2750, Denmark" SGRef: number: 4; type: "Journal Article" FEATURES Location/Qualifiers source 1..7207 /mol_type="other DNA" /organism="synthetic DNA construct" LTR 1..634 /label="3' LTR" /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label="HIV-1 Psi" /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1299..1532 /label="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1717..1761 /label="gp41 peptide" /note="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" CDS 1910..1951 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" /label="Protein Tat" enhancer 2040..2343 /label="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 2344..2547 /label="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" misc_feature 2618..2671 /label="multiple cloning site" /note="multiple cloning site" promoter complement(2675..2693) /label="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 2721..3309 /label="WPRE" /note="woodchuck hepatitis virus posttranscriptional regulatory element" LTR 3514..3746 /label="3' LTR (Delta-U3)" /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" rep_origin complement(4778..5366) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(5540..6397) /label="AmpR" /note="beta-lactamase" promoter complement(6398..6502) /label="AmpR promoter" polyA_signal 6906..7040 /label="SV40 poly(A) signal" /note="SV40 polyadenylation signal"