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:
- pBAD18-His6-hTERT-LPTS-L
- 载体抗性:
- Ampicillin
- 载体长度:
- 9120 bp
- 载体类型:
- Expression vector
- 复制子:
- ori
- 载体来源:
- Hansen DT, Thiyagarajan T, Larson AC, Hansen JL.
- 启动子:
- araBAD
pBAD18-His6-hTERT-LPTS-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。
pBAD18-His6-hTERT-LPTS-L 载体序列
LOCUS V009301 9120 bp DNA circular SYN 17-DEC-2018
DEFINITION Exported.
ACCESSION V009301
VERSION V009301
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9120)
AUTHORS Hansen DT, Thiyagarajan T, Larson AC, Hansen JL.
TITLE Telomerase repeat amplification protocol (TRAP) activity upon
recombinant expression and purification of human telomerase in a
bacterial system
JOURNAL Protein Expr. Purif. (2016) In press
PUBMED 26965413
REFERENCE 2 (bases 1 to 9120)
AUTHORS Hansen DT, Hansen JL.
TITLE Direct Submission
JOURNAL Submitted (17-DEC-2015) Department of Biochemistry and Molecular
Biology, Medical University of South Carolina, 173 Ashley Avenue,
Charleston, SC 29425, USA
REFERENCE 3 (bases 1 to 9120)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9120)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Protein
Expr. Purif. (2016) In press"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-DEC-2015) Department of Biochemistry and Molecular Biology,
Medical University of South Carolina, 173 Ashley Avenue, Charleston,
SC 29425, USA"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9120
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(99..974)
/label="araC"
/note="L-arabinose regulatory protein"
promoter 1001..1285
/label="araBAD promoter"
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
RBS 1312..1334
/label="RBS"
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1353..1370
/label="6xHis"
/note="6xHis affinity tag"
CDS 1389..4784
/label="hTERT"
/note="human telomerase reverse transcriptase, the
catalytic subunit of telomerase"
CDS 4868..5851
/gene="PINX1"
/label="PIN2/TERF1-interacting telomerase inhibitor 1"
/note="PIN2/TERF1-interacting telomerase inhibitor 1 from
Homo sapiens. Accession#: Q96BK5"
terminator 6072..6158
/label="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 6250..6277
/label="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter 6296..6387
/label="AmpR promoter"
CDS 6388..7245
/label="AmpR"
/note="beta-lactamase"
rep_origin 7290..7745
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
rep_origin 7856..8444
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(8630..8770)
/label="bom"
/note="basis of mobility region from pBR322"