pYBA-1352 载体 (V001935)

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

pYBA-13526366 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300oripBBR1 oriVpBBR1 RepRB 'overdrive' sequenceRB T-DNA repeatCaMV poly-A signal; derived from pRT103KS primerSK primerEGFPTMV OmegaCaMV 35S promoterNOS terminatorBlpRNOS promoterLB T-DNA repeatNeoR/KanR

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"