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:
- pCVD005
- 载体抗性:
- Kanamycin
- 载体长度:
- 3315 bp
- 载体类型:
- Broad host range vector
- 复制子:
- ori
- 载体来源:
- Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
- 启动子:
- tac
pCVD005 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。
pCVD005 载体序列
LOCUS 40924_49037 3315 bp DNA circular SYN 18-DEC-2018 DEFINITION Broad host range vector vector pCVD005, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3315) AUTHORS Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW. TITLE Broad-host-range vector system for synthetic biology and biotechnology in cyanobacteria JOURNAL Nucleic Acids Res. (2014) In press PUBMED 25074377 REFERENCE 2 (bases 1 to 3315) AUTHORS Taton A, Unglaub F, Wright N, Zeng WY, Paz Yepes J, Brahamsha B, Palenik B, Peterson T, Haerizadeh F, Golden SS, Golden JW. TITLE Direct Submission JOURNAL Submitted (17-JUN-2014) Division of Biological Sciences, University of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA REFERENCE 3 (bases 1 to 3315) TITLE Direct Submission REFERENCE 4 (bases 1 to 3315) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic Acids Res. (2014) In press" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (17-JUN-2014) Division of Biological Sciences, University of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..3315 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 293..309 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 320..338 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 343..354 /label=Factor Xa site /note="Factor Xa recognition and cleavage site" misc_feature 370..375 /label=EcoRV /note="EcoRV" misc_feature complement(373..393) /note="C2G; GC-adaptor for seamless assembly, flanking antibiotic resistance markers, functional devices or custom sequences; synthetic" misc_feature 394..399 /label=AgeI /note="AgeI" promoter 406..434 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" regulatory 447..471 /note="luxA 5-UTR; ribosomal binding site" /regulatory_class="promoter" CDS 472..1329 /label=AmpR /note="beta-lactamase" regulatory 1341..1367 /note="trpA T; trpA terminator from Lorist6 INSD accession X98450, one nt shorter than reference; Lorist6" /regulatory_class="terminator" misc_feature 1377..1382 /label=NheI /note="NheI" misc_feature complement(1383..1403) /note="GC; GC-adaptor for seamless assembly, flanking antibiotic resistance markers or Escherichia coli origins to be assembled with a cyanobacterial replicon; synthetic" misc_feature 1401..1406 /label=EcoRV /note="EcoRV" rep_origin complement(1631..2219) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2396..3187) /label=NeoR/KanR /note="aminoglycoside phosphotransferase" promoter complement(3188..3292) /label=AmpR promoter