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:
pBINPLUS
载体抗性:
Kanamycin
载体长度:
12396 bp
载体类型:
Plant Vectors
复制子:
oriV
宿主:
Plants
载体来源:
van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema
筛选标记:
Neomycin/G418(Geneticin)
拷贝数:
High copy number
启动子:
NOS
5'引物:
M13 fwd
3'引物:
M13 rev
感受态:
stbl3
培养温度:
37℃

pBINPLUS 载体图谱

pBINPLUS12396 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000oriVpBR322 ori (truncated)KanRtrfALB T-DNA repeatNOS promoterNeoR/KanRNOS terminatorCAP binding sitelac promoterlac operatorM13 revMCSM13 fwdRB T-DNA repeatoriVoriTtraJ

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。

pBINPLUS 载体序列

LOCUS       pBINPLUS.       12396 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Improved binary Agrobacterium vector for plant transformation.
ACCESSION   .
VERSION     .
KEYWORDS    pBINPLUS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12396)
  AUTHORS   van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema 
            WJ.
  TITLE     pBINPLUS: an improved plant transformation vector based on pBIN19.
  JOURNAL   Transgenic Res. 1995;4:288-90.
  PUBMED    7655517
REFERENCE   2  (bases 1 to 12396)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 12396)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Transgenic 
            Res."; date: "1995"; volume: "4"; pages: "288-90"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     This sequence was reconstructed using the original paper. The 
            orientation of the truncated pBR322 origin is uncertain.
FEATURES             Location/Qualifiers
     source          1..12396
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..616
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     rep_origin      1758..2110
                     /direction=RIGHT
                     /label=pBR322 ori (truncated)
                     /note="pBR322 ori (truncated)"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication (truncated)"
     CDS             2871..3662
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             3964..5109
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     misc_feature    6708..6732
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        7205..7388
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             7409..8200
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     terminator      8593..8845
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    9394..9415
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9430..9460
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9468..9484
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="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     9492..9508
                     /label=M13 rev
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(9534..9590)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(9605..9621)
                     /label=M13 fwd
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    9919..9943
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      10000..10624
                     /label=oriV
                     /note="incP origin of replication"
     oriT            11739..11848
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             11881..12249
                     /label=traJ
                     /note="oriT-recognizing protein"