pCXUN-HA 载体 (V008083)

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:
pCXUN-HA
载体抗性:
Kanamycin
载体长度:
12038 bp
载体类型:
Binary vector
复制子:
ori
载体来源:
Chen S, Songkumarn P, Liu J, Wang GL.
启动子:
Ubi

pCXUN-HA 载体图谱

pCXUN-HA12038 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000M13 fwdNOS terminatorccdBHAmaize ubiquitin-1 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。

pCXUN-HA 载体序列

LOCUS       40924_14045       12038 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCXUN-HA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12038)
  AUTHORS   Chen S, Songkumarn P, Liu J, Wang GL.
  TITLE     A versatile zero background T-vector system for gene cloning and 
            functional genomics
  JOURNAL   Plant Physiol. 150 (3), 1111-1121 (2009)
  PUBMED    19403729
REFERENCE   2  (bases 1 to 12038)
  AUTHORS   Chen S, Songkumarn P, Liu J, Wang G-L.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-APR-2009) Department of Plant Pathology, The Ohio 
            State University, 201 Kottman Hall, 2021 Coffey Rd, Columbus, OH 
            43210, USA
REFERENCE   3  (bases 1 to 12038)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12038)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Physiol."; date: "2009"; volume: "150"; issue: "3"; pages: 
            "1111-1121"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-APR-2009) Department of Plant Pathology, The Ohio State 
            University, 201 Kottman Hall, 2021 Coffey Rd, Columbus, OH 43210, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..12038
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     90..106
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(115..367)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             728..1030
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(1089..1115)
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     regulatory      complement(1131..3121)
                     /label=maize ubiquitin-1 promoter
                     /note="maize ubiquitin-1 promoter"
                     /regulatory_class="promoter"
     promoter        complement(1131..3121)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     primer_bind     complement(3143..3159)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3362..3386
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4685..5311
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             5748..6812
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      6881..7075
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    7419..7559
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7745..8333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8423..9214)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    9639..9663
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    complement(9741..9915)
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(9958..10980)
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     promoter        complement(11047..11724)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     protein_bind    11915..11936
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        11951..11981
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11989..12005
                     /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     12013..12029
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"