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
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"