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"