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

pHS2076577 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300KanRorimycobacterial origin; OriMKS primerBCG hsp60strong synthetic Shine Delgarno sequence; RBSAtatgagCycle 3 GFPrrnB T1 terminator

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  JOURNAL   Unpublished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  JOURNAL   Unpublished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"