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:
- pSolycp00007
- 载体抗性:
- Kanamycin
- 载体长度:
- 11812 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 宿主:
- Plants
- 载体来源:
- Aoki K, Kurabayashi A, Suda K, Odahara M, Goto DB, Shibata D.
- 启动子:
- CaMV 35S (enhanced)
pSolycp00007 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。
pSolycp00007 载体序列
LOCUS 40924_40827 11812 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pSolycp00007 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11812)
AUTHORS Aoki K, Kurabayashi A, Suda K, Odahara M, Goto DB, Shibata D.
TITLE 5'-flanking region of Solyc02g086820 fused to SlHTA6-EYFP
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 11812)
AUTHORS Aoki K, Kurabayashi A, Suda K, Odahara M, Goto DB, Shibata D.
TITLE Tomato HTA6 fused to EYFP driven by 35S omega promoter
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 11812)
AUTHORS Aoki K, Kurabayashi A, Suda K, Odahara M, Goto D, Shibata D.
TITLE Direct Submission
JOURNAL Submitted (24-NOV-2011) Contact:Koh Aoki Osaka Prefecture
University, Department of Life and Environmental Sciences; Naka-ku,
Gakuen-cho, 1-1, Sakai, Osaka 599-8531, Japan E-mail
:kaoki@plant.osakafu-u.ac.jp Phone :81-72-254-9421 Fax
:81-72-254-9918
REFERENCE 4 (bases 1 to 11812)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 11812)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(24-NOV-2011) Contact:Koh Aoki Osaka Prefecture University,
Department of Life and Environmental Sciences"; volume: " Naka-ku,
Gakuen-cho, 1-1, Sakai, Osaka 599-8531, Japan E-mail
:kaoki@plant.osakafu-u.ac.jp Phone :81-72-254-9421 Fax "; pages:
"81-72-254-991"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11812
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature complement(21..45)
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 248..264
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 345..369
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
regulatory 390..1889
/note="tomato promoter, BLAST search(vs SL2.40.fasta)
result/SL2.40ch02:44011063..44012562"
/regulatory_class="promoter"
protein_bind complement(1906..1930)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
misc_feature 1955..2392
/gene="SlHTA6"
/note="The nucleotide sequence is identical to GenBank
JN790634.1.; putative histone 2A protein"
gene 1955..2392
/gene="SlHTA6"
/label=SlHTA6
CDS 2399..3115
/label=EYFP
/note="enhanced YFP"
terminator 3135..3382
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(3404..3420)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3428..3444
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(3452..3482)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3497..3518)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3709..4386
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 4455..5243
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal 5303..5477
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature complement(5555..5579)
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 6004..6795
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 6885..7473
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(7659..7799)
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(8143..8337)
/direction=LEFT
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(8406..9470)
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(9907..10533)
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"