pCS 载体 (V008155)

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:
pCS
载体抗性:
Kanamycin
载体长度:
8420 bp
载体类型:
Binary vector
复制子:
ori
宿主:
Plants
载体来源:
Thomson JG, Cook M, Guttman M, Smith J, Thilmony R.
启动子:
CaMV 35S (enhanced)

pCS 载体图谱

pCS8420 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400LB T-DNA repeatCaMV35S polyA terminatorDihydropteroate synthase type-1pea rbcS ribulose-bisphosphate carboxylase small subunit transit peptide for plastid targetingCaMV 35S promoter (enhanced)RB T-DNA repeatpVS1 StaApVS1 RepApVS1 oriVbomoriKanR

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。

pCS 载体序列

LOCUS       V008155                 8420 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008155
VERSION     V008155
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8420)
  AUTHORS   Thomson JG, Cook M, Guttman M, Smith J, Thilmony R.
  TITLE     Novel sulI binary vectors enable an inexpensive foliar selection
            method in Arabidopsis
  JOURNAL   BMC Res Notes 4 (1), 44 (2011)
   PUBMED   21366919
REFERENCE   2  (bases 1 to 8420)
  AUTHORS   Thomson JG, Cook M, Guttman M, Smith J, Thilmony R.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-NOV-2010) Crop Improvement and Utilization Research
            Unit, USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA
REFERENCE   3  (bases 1 to 8420)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8420)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC Res
            Notes"; date: "2011"; volume: "4"; issue: "1"; pages: "44"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (09-NOV-2010) Crop Improvement and Utilization Research Unit,
            USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      complement(149..308)
                     /label="CaMV35S polyA terminator"
                     /note="CaMV35S polyA terminator"
                     /regulatory_class="terminator"
     CDS             complement(366..1202)
                     /gene="sulI"
                     /label="Dihydropteroate synthase type-1"
                     /note="Dihydropteroate synthase type-1 from Escherichia
                     coli. Accession#: P0C002"
     transit_peptide complement(1203..1304)
                     /gene="Sul1"
                     /note="pea rbcS ribulose-bisphosphate carboxylase small
                     subunit transit peptide for plastid targeting"
     promoter        complement(1311..1982)
                     /label="CaMV 35S promoter (enhanced)"
                     /note="cauliflower mosaic virus 35S promoter with a
                     duplicated enhancer region"
     misc_feature    2142..2166
                     /label="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3467..4093
                     /label="pVS1 StaA"
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             4530..5594
                     /label="pVS1 RepA"
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      5663..5857
                     /label="pVS1 oriV"
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6201..6341
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6527..7115)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7205..7996)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"