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:
- pCLEAN-G144
- 载体抗性:
- Kanamycin
- 载体长度:
- 4924 bp
- 载体类型:
- Binary vector
- 复制子:
- pSa ori
- 宿主:
- Plants
- 载体来源:
- Thole V, Worland B, Snape JW, Vain P.
- 启动子:
- CaMV 35S
pCLEAN-G144 载体图谱
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。
pCLEAN-G144 载体序列
LOCUS 40924_10991 4924 bp DNA circular SYN 17-DEC-2018
DEFINITION Binary vector pCLEAN-G144, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4924)
AUTHORS Thole V, Worland B, Snape JW, Vain P.
TITLE The pCLEAN dual binary vector system for Agrobacterium-mediated
plant transformation
JOURNAL Plant Physiol. 145 (4), 1211-1219 (2007)
PUBMED 17932303
REFERENCE 2 (bases 1 to 4924)
AUTHORS Thole V, Vain P.
TITLE Direct Submission
JOURNAL Submitted (01-OCT-2007) Department of Crop Genetics, John Innes
Centre, Colney Lane, Norwich NR4 7UH, UK
REFERENCE 3 (bases 1 to 4924)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4924)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant
Physiol."; date: "2007"; volume: "145"; issue: "4"; pages:
"1211-1219"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(01-OCT-2007) Department of Crop Genetics, John Innes Centre, Colney
Lane, Norwich NR4 7UH, UK"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4924
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(61..649)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(823..1635)
/label=KanR
/note="aminoglycoside phosphotransferase"
regulatory complement(1774..2070)
/label=soybean-derived poly(A)signal
/note="soybean-derived poly(A)signal"
/regulatory_class="terminator"
CDS complement(2106..2816)
/label=mgfp5
/note="GFP with folding enhancement mutations"
promoter complement(2915..3259)
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
rep_origin 3556..3991
/label=pSa ori
/note="origin of replication from bacterial plasmid pSa"
misc_feature 4118..4140
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA
(truncated)"
primer_bind 4305..4321
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 4331..4349
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature complement(4358..4465)
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(4478..4496)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4517..4533)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4541..4557)
/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)."
promoter complement(4565..4595)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4610..4631)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 4870..4894
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
misc_feature 4895..4918
/note="right border overdrive sequence (Peralta et al.
(1986) EMBO J 5: 1137-1142)"