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:
- pTIT-L
- 载体抗性:
- Ampicillin
- 载体长度:
- 9575 bp
- 载体类型:
- Rabies virus-derived expression vector
- 复制子:
- ori
- 载体来源:
- Finke S, Conzelmann KK.
pTIT-L 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。
pTIT-L 载体序列
LOCUS V002629 9575 bp DNA circular SYN 18-DEC-2018 DEFINITION Exported. ACCESSION V002629 VERSION V002629 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 9575) AUTHORS Finke S, Conzelmann KK. TITLE Virus promoters determine interference by defective RNAs: selective amplification of mini-RNA vectors and rescue from cDNA by a 3' copy-back ambisense rabies virus JOURNAL J. Virol. 73 (5), 3818-3825 (1999) PUBMED 10196276 REFERENCE 2 (bases 1 to 9575) AUTHORS Wickersham IR, Sullivan HA, Seung HS. TITLE Production of glycoprotein-deleted rabies viruses for monosynaptic tracing and high-level gene expression in neurons JOURNAL Unpublished REFERENCE 3 (bases 1 to 9575) AUTHORS Wickersham IR. TITLE Direct Submission JOURNAL Submitted (06-DEC-2009) Brain and Cognitive Sciences, Howard Hughes Medical Institute and Massachusetts Institute of Technology, 43 Vassar St., Cambridge, MA 02139, USA REFERENCE 4 (bases 1 to 9575) TITLE Direct Submission REFERENCE 5 (bases 1 to 9575) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Virol."; date: "1999"; volume: "73"; issue: "5"; pages: "3818-3825" SGRef: number: 2; type: "Journal Article"; journalName: "Unpublished" SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (06-DEC-2009) Brain and Cognitive Sciences, Howard Hughes Medical Institute and Massachusetts Institute of Technology, 43 Vassar St., Cambridge, MA 02139, USA" SGRef: number: 4; type: "Journal Article" FEATURES Location/Qualifiers source 1..9575 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 148..164 /label="M13 fwd" /note="common sequencing primer, one of multiple similar variants" promoter 171..189 /label="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 240..701 /label="IRES" /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 706..7086 /gene="L" /label="Large structural protein" /note="Large structural protein from Rabies virus (strain SAD B19). Accession#: P16289" terminator 7261..7308 /label="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(7331..7347) /label="SK primer" /note="common sequencing primer, one of multiple similar variants" promoter complement(7384..7402) /label="T3 promoter" /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7423..7439) /label="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind complement(7447..7463) /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(7471..7501) /label="lac promoter" /note="promoter for the E. coli lac operon" protein_bind complement(7516..7537) /label="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(7825..8413) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8587..9444) /label="AmpR" /note="beta-lactamase" promoter complement(9445..9549) /label="AmpR promoter"