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:
- pYBA-1352
- 载体长度:
- 6366 bp
- 载体类型:
- Binary vector
- 复制子:
- pBBR1 oriV
- 宿主:
- Plants
- 载体来源:
- Yao L, Yan X, Wang H, Cao M, Robaglia C.
- 启动子:
- CaMV 35S
pYBA-1352 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。
pYBA-1352 载体序列
LOCUS 40924_47803 6366 bp DNA circular SYN 18-DEC-2018 DEFINITION Binary vector pYBA-1352, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6366) AUTHORS Yao L, Yan X, Wang H, Cao M, Robaglia C. TITLE Direct Submission JOURNAL Submitted (16-NOV-2014) Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, 9 Shuguang Huayuan Zhonglu, Beijing 100097, P.R. China REFERENCE 2 (bases 1 to 6366) TITLE Direct Submission REFERENCE 3 (bases 1 to 6366) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted (16-NOV-2014) Beijing Agro-Biotechnology Research Center, Beijing Academy of Agriculture and Forestry Sciences, 9 Shuguang Huayuan Zhonglu, Beijing 100097, P.R. China" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..6366 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin complement(1..589) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" rep_origin 741..1510 /label=pBBR1 oriV /note="replication origin of the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica; requires the pBBR1 Rep protein for replication" CDS 1511..2170 /label=pBBR1 Rep /note="replication protein for the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica" misc_feature 2368..2391 /label=RB 'overdrive' sequence /note="RB 'overdrive' sequence" misc_feature complement(2392..2416) /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" misc_feature 2428..2628 /note="CaMV poly-A signal; derived from pRT103" primer_bind 2662..2678 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2712..2728) /label=SK primer /note="common sequencing primer, one of multiple similar variants" CDS complement(2729..3445) /label=EGFP /note="enhanced GFP" misc_feature complement(3451..3506) /label=TMV Omega /note="translational enhancer from the tobacco mosaic virus 5'-leader sequence (Gallie et al., 1988)" promoter complement(3517..3862) /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" terminator complement(3964..4216) /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS complement(4220..4768) /label=BlpR /note="phosphinothricin acetyltransferase" promoter complement(4777..4960) /label=NOS promoter /note="nopaline synthase promoter" misc_feature complement(5095..5119) /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS complement(5309..6100) /label=NeoR/KanR /note="aminoglycoside phosphotransferase"