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:
- pFA6a-TRP1-PGAL1-HBH
- 载体抗性:
- Ampicillin
- 载体长度:
- 4477 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Tagwerker C, Zhang H, Wang X, Larsen LS, Lathrop RH, Hatfield GW, Auer B, Huang L, Kaiser P.
- 启动子:
- GAL1
pFA6a-TRP1-PGAL1-HBH 载体载体图谱
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。
pFA6a-TRP1-PGAL1-HBH 载体载体序列
LOCUS 40924_19591 4477 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector pFA6a-TRP1-PGAL1-HBH, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4477) AUTHORS Tagwerker C, Zhang H, Wang X, Larsen LS, Lathrop RH, Hatfield GW, Auer B, Huang L, Kaiser P. TITLE HB tag modules for PCR-based gene tagging and tandem affinity purification in Saccharomyces cerevisiae JOURNAL Yeast 23 (8), 623-632 (2006) PUBMED 16823883 REFERENCE 2 (bases 1 to 4477) AUTHORS Kaiser P, Tagwerker C. TITLE Direct Submission JOURNAL Submitted (16-FEB-2006) Biological Chemistry, UC Irvine, 19182 Jamboree Blvd RM H136 Plumwood Bldg, Irvine, CA 92697, USA REFERENCE 3 (bases 1 to 4477) AUTHORS Kaiser P, Tagwerker C. TITLE Direct Submission JOURNAL Submitted (21-AUG-2009) Biological Chemistry, UC Irvine, 19182 Jamboree Blvd RM H136 Plumwood Bldg, Irvine, CA 92697, USA REFERENCE 4 (bases 1 to 4477) TITLE Direct Submission REFERENCE 5 (bases 1 to 4477) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Yeast"; date: "2006"; volume: "23"; issue: "8"; pages: "623-632" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (16-FEB-2006) Biological Chemistry, UC Irvine, 19182 Jamboree Blvd RM H136 Plumwood Bldg, Irvine, CA 92697, USA" COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (21-AUG-2009) Biological Chemistry, UC Irvine, 19182 Jamboree Blvd RM H136 Plumwood Bldg, Irvine, CA 92697, USA" COMMENT SGRef: number: 4; type: "Journal Article" COMMENT On Aug 21, 2009 this sequence version replaced DQ407931.1. FEATURES Location/Qualifiers source 1..4477 /mol_type="other DNA" /organism="synthetic DNA construct" promoter complement(1..19) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" promoter 365..469 /label=AmpR promoter CDS 470..1327 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1501..2089 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 2347..2365 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 2438..2446 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" CDS complement(2523..3194) /codon_start=1 /label=TRP1 /note="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" promoter complement(3195..3342) /label=TRP1 promoter promoter 3437..3878 /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" CDS 3925..3942 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" CDS 4171..4188 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" terminator 4215..4402 /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene"