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"