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:
- pDOE-12
- 载体抗性:
- Kanamycin
- 载体长度:
- 11595 bp
- 载体类型:
- Binary vector
- 复制子:
- ori
- 载体来源:
- Gookin TE, Assmann SM.
- 启动子:
- MAS
pDOE-12 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。
pDOE-12 载体序列
LOCUS 40924_14965 11595 bp DNA circular SYN 17-DEC-2018
DEFINITION Binary vector pDOE-12, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11595)
AUTHORS Gookin TE, Assmann SM.
TITLE Significant reduction of BiFC non-specific assembly facilitates in
planta assessment of heterotrimeric G-protein interactors
JOURNAL Plant J. (2014) In press
PUBMED 25187041
REFERENCE 2 (bases 1 to 11595)
AUTHORS Gookin TE, Assmann SM.
TITLE Direct Submission
JOURNAL Submitted (09-SEP-2014) Department of Biology, The Pennsylvania
State University, 208 Mueller Laboratory, University Park, PA 16802,
USA
REFERENCE 3 (bases 1 to 11595)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11595)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant J.
(2014) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-SEP-2014) Department of Biology, The Pennsylvania State
University, 208 Mueller Laboratory, University Park, PA 16802, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11595
/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"
misc_feature 99..104
/label=non-unique SacI
/note="non-unique SacI"
terminator complement(111..363)
/label=MAS terminator
/note="mannopine synthase terminator"
misc_feature complement(373..406)
/label=BAR gene remnant
/note="BAR gene remnant"
misc_feature 407..412
/label=MCS2 KpnI site
/note="MCS2 KpnI site"
CDS complement(416..1129)
/label=mTurquoise2
/note="enhanced monomeric variant of CFP (Goedhart et al.,
2012)"
sig_peptide complement(1130..1234)
/gene="XT-Golgi-mTq2"
/note="exclusive localization to the medial Golgi
cisternae; from Arabidopsis AtXylT/XYLT
(beta-1,2-xylosyltransferase, At5g55500); characterized in
PubMed ID 12943552; confirmed by Saint-Jore-Dupas et al.
(Plant Cell 2006;18;3182-3200)"
misc_feature 1235..1240
/label=MCS2 NruI site
/note="MCS2 NruI site"
regulatory complement(1238..1618)
/note="mas1'-2' mannopine synthase promoter; mas2' drives
MCS2 ORF; mas1' truncated by 96 nt; enhancer elements, TATA
box, and start codon missing"
/regulatory_class="promoter"
promoter complement(1238..1618)
/label=MAS promoter
/note="mannopine synthase promoter (Velten et al., 1984)"
regulatory 1631..2268
/note="UBQ10; Arabidopsis ubiquitin10 promoter"
/regulatory_class="promoter"
misc_difference 1784
/replace="g"
/label=killed PmlI (CACcTG)
/note="killed PmlI (CACcTG)"
misc_feature 1965..2268
/label=cryptic intron from Norris et al. 1993
/note="cryptic intron from Norris et al. 1993"
misc_difference 2197
/replace="t"
/label=killed XbaI (aCTAGA)
/note="killed XbaI (aCTAGA)"
primer_bind 2269..2293
/label=pDOE mcs1F with XhoI
/note="pDOE mcs1F with XhoI"
misc_feature 2281..2297
/label=XhoSac linker
/note="XhoSac linker"
misc_feature 2308..2362
/label=TMV Omega
/note="translational enhancer from the tobacco mosaic virus
5'-leader sequence (Gallie et al., 1988)"
CDS 2364..3047
/codon_start=1
/gene="NmVen210-X"
/product="NmVenus210-mcs1 fusion protein"
/label=NmVen210-X
/protein_id="AIN46649.1"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKGASGGGSGSMGR
ALGTS"
gene 2364..3047
/gene="NmVen210-X"
/label=NmVen210-X
misc_feature 2364..2993
/gene="NmVen210-X"
/label=NmVenus210 specific sequence
/note="NmVenus210 specific sequence"
CDS 2364..2882
/codon_start=1
/product="N-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VN173
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIE"
misc_difference 2982..2984
/gene="NmVen210-X"
/label=A206K monomerizing mutation
/note="A206K monomerizing mutation"
misc_feature 2994..3047
/label=MCS1 front end
/note="MCS1 front end"
misc_feature 3048..3069
/label=MCS1 back end
/note="MCS1 back end"
terminator 3080..3787
/label=OCS terminator
/note="octopine synthase terminator"
regulatory 3808..4445
/note="UBQ10; ubiquitin10 promoter"
/regulatory_class="promoter"
misc_difference 3961
/replace="g"
/label=killed PmlI (CACcTG)
/note="killed PmlI (CACcTG)"
misc_difference 4374
/replace="t"
/label=killed XbaI (aCTAGA)
/note="killed XbaI (aCTAGA)"
primer_bind 4446..4470
/label=pDOE mcs3bF with Bsu36I
/note="pDOE mcs3bF with Bsu36I"
misc_feature 4453..4468
/label=Bsu361 linker
/note="Bsu361 linker"
regulatory 4469..4534
/label=TMV-Omega enhancer
/note="TMV-Omega enhancer"
/regulatory_class="enhancer"
misc_feature 4479..4533
/label=TMV Omega
/note="translational enhancer from the tobacco mosaic virus
5'-leader sequence (Gallie et al., 1988)"
misc_feature 4538..4624
/gene="CVen210-SFS-X"
/label=CVenus210 specific sequence
/note="CVenus210 specific sequence"
CDS 4637..4660
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
CDS 4670..4693
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 4700..4723
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
misc_feature 4739..4774
/label=MCS3 front end
/note="MCS3 front end"
misc_feature 4775..4792
/label=MCS3 back end
/note="MCS3 back end"
terminator 4794..5046
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 5056..5072
/label=remnant from pFGC5941
/note="remnant from pFGC5941"
primer_bind complement(5097..5113)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 5316..5340
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 6641..7267
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
CDS 7699..8769
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
rep_origin 8838..9032
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 9376..9516
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(9702..10290)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10380..11171)
/label=KanR
/note="aminoglycoside phosphotransferase"