pCAMBIA1300-VE3-2S 载体 (V008733)

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:
pCAMBIA1300-VE3-2S
载体抗性:
Kanamycin
载体长度:
11369 bp
载体类型:
Binary vector
复制子:
ori
宿主:
Plants
载体来源:
Shi XG, Zhuang WJ.
启动子:
CaMV 35S (enhanced)

pCAMBIA1300-VE3-2S 载体图谱

pCAMBIA1300-VE3-2S11369 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000NOS terminator2-methyl-6-phytyl-1,4-hydroquinone methyltransferase, chloroplasticBrassica napus 2S seed storage protein promoterM13 fwdRB T-DNA repeatpVS1 StaApVS1 RepApVS1 oriVbomoriKanRLB T-DNA repeatCaMV poly(A) signalHygRCaMV 35S promoter (enhanced)CAP binding sitelac promoterlac operatorM13 rev

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。

pCAMBIA1300-VE3-2S 载体序列

LOCUS       V008733                11369 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008733
VERSION     V008733
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11369)
  AUTHORS   Shi XG, Zhuang WJ.
  TITLE     Binary vector pCAMBIA1300-VE3-2S for making MPBQ/MSBQ
            methyltransferase over expression in seed
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11369)
  AUTHORS   Zhuang WJ, Shi XG.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-OCT-2008) Crop Science Department, Oil Crop
            Institution, Fujian Agriculture, Jinshan, Fuzhou, Fujian 350002,
            P.R. China
REFERENCE   3  (bases 1 to 11369)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11369)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (08-OCT-2008) Crop Science Department, Oil Crop Institution, Fujian
            Agriculture, Jinshan, Fuzhou, Fujian 350002, P.R. China"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(8..260)
                     /label="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(280..1293)
                     /gene="VTE3"
                     /label="2-methyl-6-phytyl-1,4-hydroquinone
                     methyltransferase, chloroplastic"
                     /note="2-methyl-6-phytyl-1,4-hydroquinone
                     methyltransferase, chloroplastic from Arabidopsis thaliana.
                     Accession#: Q9LY74"
     regulatory      1305..2449
                     /label="Brassica napus 2S seed storage protein promoter"
                     /note="Brassica napus 2S seed storage protein promoter"
                     /regulatory_class="promoter"
     primer_bind     complement(2471..2487)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    2690..2714
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4014..4640
                     /label="pVS1 StaA"
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             5077..6141
                     /label="pVS1 RepA"
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      6210..6404
                     /label="pVS1 oriV"
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6748..6888
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7074..7662)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7752..8543)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     misc_feature    8968..8992
                     /label="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(9070..9244)
                     /label="CaMV poly(A) signal"
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(9287..10309)
                     /label="HygR"
                     /note="aminoglycoside phosphotransferase from E. coli"
     promoter        complement(10377..11054)
                     /label="CaMV 35S promoter (enhanced)"
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     protein_bind    11245..11266
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        11281..11311
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    11319..11335
                     /label="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     11343..11359
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"