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:
- pBINPLUS
- 载体抗性:
- Kanamycin
- 载体长度:
- 12396 bp
- 载体类型:
- Plant Vectors
- 复制子:
- oriV
- 宿主:
- Plants
- 载体来源:
- van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema
- 筛选标记:
- Neomycin/G418(Geneticin)
- 拷贝数:
- High copy number
- 启动子:
- NOS
- 5'引物:
- M13 fwd
- 3'引物:
- M13 rev
- 感受态:
- stbl3
- 培养温度:
- 37℃
pBINPLUS 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。
pBINPLUS 载体序列
LOCUS pBINPLUS. 12396 bp DNA circular SYN 01-JAN-1980 DEFINITION Improved binary Agrobacterium vector for plant transformation. ACCESSION . VERSION . KEYWORDS pBINPLUS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12396) AUTHORS van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema WJ. TITLE pBINPLUS: an improved plant transformation vector based on pBIN19. JOURNAL Transgenic Res. 1995;4:288-90. PUBMED 7655517 REFERENCE 2 (bases 1 to 12396) TITLE Direct Submission REFERENCE 3 (bases 1 to 12396) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Transgenic Res."; date: "1995"; volume: "4"; pages: "288-90" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT This sequence was reconstructed using the original paper. The orientation of the truncated pBR322 origin is uncertain. FEATURES Location/Qualifiers source 1..12396 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 6..616 /label=oriV /note="origin of replication for the bacterial F plasmid" rep_origin 1758..2110 /direction=RIGHT /label=pBR322 ori (truncated) /note="pBR322 ori (truncated)" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication (truncated)" CDS 2871..3662 /label=KanR /note="aminoglycoside phosphotransferase" CDS 3964..5109 /label=trfA /note="trans-acting replication protein that binds to and activates oriV" misc_feature 6708..6732 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" promoter 7205..7388 /label=NOS promoter /note="nopaline synthase promoter" CDS 7409..8200 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" terminator 8593..8845 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" protein_bind 9394..9415 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 9430..9460 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9468..9484 /label=lac repressor encoded by lacI binding site /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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 9492..9508 /label=M13 rev /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" misc_feature complement(9534..9590) /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(9605..9621) /label=M13 fwd /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" misc_feature 9919..9943 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" rep_origin 10000..10624 /label=oriV /note="incP origin of replication" oriT 11739..11848 /label=oriT /note="incP origin of transfer" CDS 11881..12249 /label=traJ /note="oriT-recognizing protein"