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:
pBin19
载体抗性:
Kanamycin
载体长度:
11777 bp
载体类型:
Plant Vectors
复制子:
oriV
宿主:
Plants
载体来源:
Bevan M.
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

pBin19 载体图谱

pBin1911777 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105001100011500oriVIS1KanRtrfALB T-DNA repeatM13 fwdMCSM13 revlac operatorlac promoterCAP binding siteNOS terminatorNeoR/KanRNOS promoterRB 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。

pBin19 载体序列

LOCUS       pBin19.       11777 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Binary Agrobacterium vector for plant transformation. Also known as 
            pBIN19 or BIN19 or Bin19 or Bin 19.
ACCESSION   .
VERSION     .
KEYWORDS    pBin19
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11777)
  AUTHORS   Bevan M.
  TITLE     Binary Agrobacterium vectors for plant transformation.
  JOURNAL   Nucleic Acids Res. 1984;12:8711-21.
  PUBMED    6095209
REFERENCE   2  (bases 1 to 11777)
  AUTHORS   Frisch DA, Harris-Haller LW, Yokubaitis NT, Thomas TL, Hardin SH, 
            Hall TC.
  TITLE     Complete sequence of the binary vector Bin 19.
  JOURNAL   Plant Mol. Biol. 1995;27:405-9.
  PUBMED    7888628
REFERENCE   3  (bases 1 to 11777)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11777)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "1984"; volume: "12"; pages: "8711-21"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Plant Mol. 
            Biol."; date: "1995"; volume: "27"; pages: "405-9"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      6..616
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     mobile_element  1317..2084
                     /mobile_element_type="insertion sequence:IS1"
                     /label=insertion sequence:IS1
                     /note="IS1"
                     /note="prokaryotic transposable element"
     CDS             2276..3067
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             3369..4514
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     misc_feature    6113..6137
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     6755..6771
                     /label=M13 fwd
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6772..6828
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(6841..6857)
                     /label=M13 rev
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6865..6881
                     /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)."
     promoter        complement(6889..6919)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6934..6955)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     terminator      complement(7504..7756)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(8149..8937)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     promoter        complement(8961..9144)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    9300..9324
                     /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            11120..11229
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             11262..11630
                     /label=traJ
                     /note="oriT-recognizing protein"