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:
- pSEVA451
- 载体抗性:
- Streptomycin
- 载体长度:
- 5334 bp
- 载体类型:
- Cloning vector
- 复制子:
- RSF1010 oriV
- 载体来源:
- Silva-Rocha R, Martinez-Garcia E, Chavarria M, Calles B, Arce-Rodriguez A, de las Heras A, Paez-Espino D, Durante-Rodriguez G, Kim J, Nikel PI, Platero R, de Lorenzo V.
- 启动子:
- lac UV5
pSEVA451 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。
pSEVA451 载体序列
LOCUS 40924_39543 5334 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pSEVA451, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5334)
AUTHORS Silva-Rocha R, Martinez-Garcia E, Chavarria M, Calles B,
Arce-Rodriguez A, de las Heras A, Paez-Espino D, Durante-Rodriguez
G, Kim J, Nikel PI, Platero R, de Lorenzo V.
TITLE The Standard European Vector Architecture (SEVA): a coherent
platform for analysis and deployment of complex prokaryotic
phenotypes
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5334)
AUTHORS Silva-Rocha R, Martinez-Garcia E, Chavarria M, Calles B,
Arce-Rodriguez A, de las Heras A, Paez-Espino D, Durante-Rodriguez
G, Kim J, Nikel PI, Platero R, de Lorenzo V.
TITLE Direct Submission
JOURNAL Submitted (28-AUG-2012) Systems Biology Program, Centro Nacional de
Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid,
Madrid 28049, Spain
REFERENCE 3 (bases 1 to 5334)
AUTHORS Silva-Rocha R, Martinez-Garcia E, Chavarria M, Calles B,
Arce-Rodriguez A, de las Heras A, Paez-Espino D, Durante-Rodriguez
G, Kim J, Nikel PI, Platero R, de Lorenzo V.
TITLE Direct Submission
JOURNAL Submitted (30-SEP-2014) Systems Biology Program, Centro Nacional de
Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid,
Madrid 28049, Spain
REFERENCE 4 (bases 1 to 5334)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 5334)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(28-AUG-2012) Systems Biology Program, Centro Nacional de
Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid,
Madrid 28049, Spain"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(30-SEP-2014) Systems Biology Program, Centro Nacional de
Biotecnologia-CSIC, C/ Darwin 3, Campus de Cantoblanco, Madrid,
Madrid 28049, Spain"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Assembly Method :: ApE v. v2.0.40
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
On Sep 30, 2014 this sequence version replaced JX560340.1.
FEATURES Location/Qualifiers
source 1..5334
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 9..32
/label=R24
/note="R24"
misc_feature 56..112
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(121..144)
/label=F24
/note="F24"
terminator 155..249
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 485..1273
/codon_start=1
/label=SmR
/note="aminoglycoside adenylyltransferase (Murphy, 1985)"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
oriT 1422..1530
/label=oriT
/note="incP origin of transfer"
CDS complement(1544..2392)
/codon_start=1
/label=RSF1010 RepC
/note="replication protein C of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/translation="VVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
AG"
CDS complement(2382..3218)
/codon_start=1
/label=RSF1010 RepA
/note="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
CDS complement(3732..4700)
/codon_start=1
/label=RSF1010 RepB
/note="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
promoter complement(4724..4754)
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
rep_origin 4810..5204
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
terminator complement(5242..5328)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"