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:
- pX6-GFP
- 载体抗性:
- Streptomycin
- 载体长度:
- 14103 bp
- 载体类型:
- Plant DNA excision vector
- 复制子:
- ori
- 宿主:
- Plants
- 载体来源:
- Zuo J, Niu QW, Moller SG, Chua NH.
- 启动子:
- NOS
pX6-GFP 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。
pX6-GFP 载体序列
LOCUS 40924_47053 14103 bp DNA circular SYN 18-DEC-2018
DEFINITION Plant DNA excision vector pX6-GFP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14103)
AUTHORS Zuo J, Niu QW, Moller SG, Chua NH.
TITLE Chemical-regulated, site-specific DNA excision in transgenic plants
JOURNAL Nat. Biotechnol. 19 (2), 157-161 (2001)
PUBMED 11175731
REFERENCE 2 (bases 1 to 14103)
AUTHORS Zuo J, Chua N-H.
TITLE Direct Submission
JOURNAL Submitted (20-DEC-2000) Laboratory of Plant Molecular Biology, The
Rockefeller University, 1230 York Avenue, New York, NY 10021, USA
REFERENCE 3 (bases 1 to 14103)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 14103)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
Biotechnol."; date: "2001"; volume: "19"; issue: "2"; pages:
"157-161"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(20-DEC-2000) Laboratory of Plant Molecular Biology, The Rockefeller
University, 1230 York Avenue, New York, NY 10021, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14103
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(1..25)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
promoter 216..262
/label=minimal CaMV 35S promoter
/note="minimal 35S promoter from cauliflower mosaic virus"
protein_bind complement(280..313)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS 332..583
/label=LexA BD
/note="DNA-binding domain of E. coli LexA"
CDS 599..832
/label=VP16 AD
/note="transcriptional activation domain of herpes simplex
virus protein VP16 (Triezenberg et al., 1988; Cousens et
al., 1989)"
CDS 842..1777
/label=ERT2
/note="mutated ligand-binding domain of the human estrogen
receptor (Feil et al., 1997)"
3'UTR 1797..1890
/label=rat glucocorticoid receptor
/note="rat glucocorticoid receptor"
regulatory 1916..2211
/label=pea rbcS E9
/note="pea rbcS E9"
/regulatory_class="terminator"
promoter 2368..2551
/label=NOS promoter
/note="nopaline synthase promoter"
CDS 2575..3363
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
misc_feature 3368..3739
/label=derived from pBI101
/note="derived from pBI101"
terminator 3756..4008
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 4099..4249
/label=LexA operator
/note="LexA operator"
regulatory 4329..4387
/label=35S promoter
/note="35S promoter"
/regulatory_class="promoter"
promoter 4331..4377
/label=minimal CaMV 35S promoter
/note="minimal 35S promoter from cauliflower mosaic virus"
5'UTR 4390..4408
/label=cre
/note="cre"
CDS join(4409..4840,5011..5610)
/codon_start=1
/product="cre"
/label=cre
/protein_id="AAK08508.1"
/translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
DGD"
intron 4841..5010
/number=5
/note="Arabidopsis KOR1 gene"
3'UTR 5611..5620
/label=cre
/note="cre"
regulatory 5624..6036
/label=Nos
/note="Nos"
/regulatory_class="terminator"
terminator 5788..6040
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
protein_bind complement(6041..6074)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS 6101..6814
/label=GFP (S65T)
/note="S65T variant of Aequorea victoria green fluorescent
protein (Heim et al., 1995)"
regulatory 6830..7299
/label=3A
/note="3A"
/regulatory_class="terminator"
misc_feature 7305..7505
/label=derived from pUC9
/note="derived from pUC9"
primer_bind complement(7331..7347)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 7506..7562
/label=derived from pBI101
/note="derived from pBI101"
misc_feature complement(7569..7593)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 8121..8909
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
rep_origin 9156..9744
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(9930..10070)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(10414..10608)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(10677..11741)
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(12178..12804)
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"