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。

The pBI121 is a binary Agrobacterium vector equipped with a GUS reporter gene designed for facilitating plant transformation.

载体名称:
pBI121
载体抗性:
Kanamycin
载体长度:
14758 bp
载体类型:
Plant Vectors
复制子:
oriV
宿主:
Plants
载体来源:
Chen PY, Wang CK, Soong SC, To KY.
筛选标记:
Neomycin/G418(Geneticin)
拷贝数:
High copy number
启动子:
CaMV35S(long)
5'引物:
M13 fwd
3'引物:
M13 rev
感受态:
stbl3
培养温度:
37℃

pBI121 载体图谱

pBI12114758 bp700140021002800350042004900560063007000770084009100980010500112001190012600133001400014700traJoriTColE1 ori; similar to sequence in GenBank Accession Number V00268TetRRB T-DNA repeatNOS promoterNeoR/KanRoriVCAP binding sitelac promoterlac operatorM13 revCaMV 35S promoterGUSNOSM13 fwdLB T-DNA repeatsimilar to tetA in GenBank Accession Number L13842trfAKanRIS1similarity to NPT III gene in GenBank Accession Number V01547similar to kilA in GenBank Accession Number M62846oriV

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。

pBI121 载体序列

LOCUS       Exported               14758 bp DNA     circular SYN 03-SEP-2024
DEFINITION  Binary vector pBI121, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14758)
  AUTHORS   Chen PY, Wang CK, Soong SC, To KY.
  TITLE     Complete sequence of the binary vector pBI121 and its application in
            cloning T-DNA insertion from transgenic plants
  JOURNAL   Mol. Breed. 11, 287-293 (2003)
REFERENCE   2  (bases 1 to 14758)
  AUTHORS   To KY.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-FEB-2002) Institute of BioAgricultural Sciences, 
            Academia Sinica, Taipei 11529, Taiwan
REFERENCE   3  (bases 1 to 14758)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 14758)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 14758)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Breed.
            11, 287-293 (2003)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (20-FEB-2002) Institute of BioAgricultural Sciences, Academia
            Sinica, Taipei 11529, Taiwan"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..14758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(148..516)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(549..658)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      complement(790..1168)
                     /direction=LEFT
                     /note="ColE1 ori; similar to sequence in GenBank Accession 
                     Number V00268"
     CDS             complement(1342..1989)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     misc_feature    complement(2454..2478)
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        2634..2817
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             2841..3629
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     rep_origin      complement(4135..4760)
                     /direction=LEFT
                     /label=oriV
                     /note="incP origin of replication"
     protein_bind    4823..4844
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4859..4889
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4897..4913
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     4921..4937
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5461..5806
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             5845..7653
                     /label=GUS
                     /note="beta-glucuronidase"
     regulatory      7727..7979
                     /label=NOS
                     /note="NOS"
                     /regulatory_class="terminator"
     terminator      7727..7979
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(7988..8004)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(8622..8646)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(9156..10198)
                     /label=similar to tetA in GenBank Accession Number L13842
                     /note="similar to tetA in GenBank Accession Number L13842"
     CDS             complement(10245..11390)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     CDS             complement(11692..12483)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     mobile_element  12675..13442
                     /label=IS1
                     /note="prokaryotic transposable element"
     misc_feature    complement(13444..13794)
                     /note="similarity to NPT III gene in GenBank Accession
                     Number V01547"
     misc_feature    complement(13795..14066)
                     /label=similar to kilA in GenBank Accession Number M62846
                     /note="similar to kilA in GenBank Accession Number M62846"
     rep_origin      complement(14143..14753)
                     /direction=LEFT
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"