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。
pSET4s, pSET5s, and pSET6s are three thermosensitive (Ts) suicide vectors used for gene replacement in Streptococcus suis. These vectors could be propagated at 37°C in Escherichia coli, but their replication was blocked above 37°C in S. suis. orfC, replication regulatory protein; repATs, thermosensitive replication initiation–termination protein.
- 载体名称:
- pSET4s
- 载体抗性:
- Spectinomycin
- 载体长度:
- 4505 bp
- 载体类型:
- Thermosensitive suicide vector
- 复制子:
- ori
- 载体来源:
- Takamatsu D, Osaki M, Sekizaki T.
- 感受态:
- DH5alpha
- 培养温度:
- 37℃
pSET4s 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。
pSET4s 载体序列
LOCUS Exported 4505 bp DNA circular SYN 02-SEP-2024
DEFINITION Thermosensitive suicide vector pSET4s DNA, complete sequence.
ACCESSION AB055650
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4505)
AUTHORS Takamatsu D, Osaki M, Sekizaki T.
TITLE Thermosensitive suicide vectors for gene replacement in
Streptococcus suis
JOURNAL Plasmid 46 (2), 140-148 (2001)
PUBMED 11591139
REFERENCE 2 (bases 1 to 4505)
AUTHORS Takamatsu D, Osaki M, Sekizaki T.
TITLE Direct Submission
JOURNAL Submitted (08-FEB-2001) Daisuke Takamatsu, National Institute of
Animal Health, Laboratory of Molecular Bacteriology; Kannondai
3-1-1, Tsukuba, Ibaraki 305-0856, Japan
(E-mail:p1013dt@niah.affrc.go.jp, Tel:81-298-38-7743,
Fax:81-298-38-7743)
REFERENCE 3 (bases 1 to 4505)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
date: "2001"; volume: "46"; issue: "2"; pages: "140-148"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(08-FEB-2001) Daisuke Takamatsu, National Institute of Animal
Health, Laboratory of Molecular Bacteriology"; volume: " Kannondai
3-1-1, Tsukuba, Ibaraki 305-0856, Japan
(E-mail:p1013dt@niah.affrc.go.jp, Tel:81-298-38-7743, Fax"; pages:
"81-298-38-7743"
FEATURES Location/Qualifiers
source 1..4505
/mol_type="other DNA"
/db_xref="taxon:152114"
/organism="Thermosensitive suicide vector pSET4s"
gene 528..689
/gene="orfC"
/label=orfC
CDS 528..689
/codon_start=1
/gene="orfC"
/product="ORFC"
/label=orfC
/protein_id="BAB64880.1"
/translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
RKKSEQKK"
gene 756..1454
/gene="repAts"
/label=repAts
CDS 756..1454
/codon_start=1
/gene="repAts"
/product="RepAts"
/label=repAts
/protein_id="BAB64881.1"
/translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
VLDAETGEIK"
gene complement(1803..2126)
/gene="lacZ'"
/label=lacZ'
CDS complement(1803..2126)
/codon_start=1
/gene="lacZ'"
/product="LacZ'"
/label=lacZ'
/protein_id="BAB64882.1"
/translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
DAA"
primer_bind 2036..2052
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 2053..2109
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(2122..2138)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2146..2162
/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(2170..2200)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2215..2236
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(2524..3112)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
gene 3637..4404
/gene="spc"
/label=spc
CDS 3637..4404
/codon_start=1
/gene="spc"
/product="spectinomycin adenyltransferase"
/label=spc
/protein_id="BAB64883.1"
/translation="MRRIYLNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNS
DLDFLVVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQ
EFIYGEWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVR
RAIMDSSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERI
LLAVRSYLGENIEWTNENVNLTINYLNNRLKKL"