pYeaR-Bxb1 载体 (V001904)

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:
pYeaR-Bxb1
载体抗性:
Chloramphenicol
载体长度:
3895 bp
载体类型:
Controller 1 vector
复制子:
ori
载体来源:
Courbet A, Endy D, Renard E, Molina F, Bonnet J.

pYeaR-Bxb1 载体图谱

pYeaR-Bxb13895 bp60012001800240030003600BioBrick prefixpYeaRsTRSV HHRzBxb1 integraseBioBrick suffixhis operon terminatororilambda t0 terminatorCmRcat promoterbacterial 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。

pYeaR-Bxb1 载体序列

LOCUS       40924_47993        3895 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Controller 1 vector pYeaR-Bxb1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3895)
  AUTHORS   Courbet A, Endy D, Renard E, Molina F, Bonnet J.
  TITLE     Detection of pathological biomarkers in human clinical samples via 
            amplifying genetic switches and logic gates
  JOURNAL   Sci Transl Med 7 (289), 289RA83 (2015)
  PUBMED    26019219
REFERENCE   2  (bases 1 to 3895)
  AUTHORS   Courbet A, Molina F, Endy D, Bonnet J.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUL-2014) SysDiag CNRS UMR3145, CNRS, 1682 Rue de la 
            Valsiere, Montpellier, Herault 34184, France
REFERENCE   3  (bases 1 to 3895)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3895)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Transl 
            Med"; date: "2015"; volume: "7"; issue: "289"; pages: "289RA83"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-JUL-2014) SysDiag CNRS UMR3145, CNRS, 1682 Rue de la Valsiere, 
            Montpellier, Herault 34184, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     regulatory      23..122
                     /label=pYeaR
                     /note="pYeaR"
                     /regulatory_class="promoter"
     misc_RNA        136..186
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             244..1785
                     /codon_start=1
                     /product="Bxb1 integrase"
                     /label=Bxb1 integrase
                     /protein_id="AKC96464.1"
                     /translation="MRALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAG
                     SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMSTGAANDENYAAHL"
     misc_feature    1789..1809
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1810..1869
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E.
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     rep_origin      complement(2120..2708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(2796..2890)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2914..3570)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3571..3673)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     terminator      complement(3850..3893)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"