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:
- pHS207
- 载体抗性:
- Kanamycin
- 载体长度:
- 6577 bp
- 载体类型:
- Expression vector
- 复制子:
- ori
- 载体来源:
- Bourn WR, Shephard E, Powels R, Stutz H, Douglas N, Jaffray A, Williamson A-L.
pHS207 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。
pHS207 载体序列
LOCUS 40924_25056 6577 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pHS207, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6577)
AUTHORS Bourn WR, Shephard E, Powels R, Stutz H, Douglas N, Jaffray A,
Williamson A-L.
TITLE The use of guided evolution to create a stable, immunogenic HIV-1
antigen expressing recombinant BCG
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6577)
AUTHORS Shephard E, Bourn WR, Stutz H, Johnston N, van Harmelen JH,
Williamson C, Douglass N, Bowers D, Galant S, Isaacs Z, Binder A,
Williamson A-L.
TITLE Priming with a rBCG or DNA vaccine expressing HIV-I subtype C Gag
elicits comparable CD8+ T cell responses when boosted by a rMVA
expressing GagC
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 6577)
AUTHORS Stutz H, Bourn WR.
TITLE Direct Submission
JOURNAL Submitted (10-JAN-2007) Medical Virology and Institute of Infectious
Disease and Molecular Medicine, University of Cape Town, Anzio Road
Observatory, Cape Town 7925, South Africa
REFERENCE 4 (bases 1 to 6577)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6577)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(10-JAN-2007) Medical Virology and Institute of Infectious Disease
and Molecular Medicine, University of Cape Town, Anzio Road
Observatory, Cape Town 7925, South Africa"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6577
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 120..932
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 1267..1855
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
rep_origin 2215..4799
/note="mycobacterial origin; OriM"
primer_bind 4829..4845
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
regulatory 4857..4943
/label=BCG hsp60
/note="BCG hsp60"
/regulatory_class="promoter"
regulatory 4967..4973
/note="strong synthetic Shine Delgarno sequence; RBSA"
/regulatory_class="ribosome_binding_site"
gene 5010..5372
/gene="tat"
/label=tat
/note="shuffled BCG codon optimized tat gene from HIV-1
subtype C isolate"
gene 5382..5719
/gene="gag"
/label=gag
/note="truncated BCG codon optimized gag gene from HIV-1
subtype C isolate"
misc_feature 5720..5752
/label=Gag-GFP linker sequence
/note="Gag-GFP linker sequence"
CDS 5747..6460
/label=Cycle 3 GFP
/note="Cycle 3 GFP (Crameri et al., 1996)"
terminator 6495..6541
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"