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。
Suicide Plasmid
- 载体名称:
- pEX18Tc
- 载体抗性:
- Tetracycline
- 载体长度:
- 6369 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP.
- 启动子:
- sacB
- 克隆方法:
- restriction endonuclease,
- 5'引物:
- M13F (-47) CGCCAGGGTTTTCCCAGTCACGAC
- 3'引物:
- M13R (-48) AGCGGATAACAATTTCACACAGGA
pEX18Tc 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。
pEX18Tc 载体序列
LOCUS Exported 6369 bp DNA circular SYN 02-SEP-2024
DEFINITION Cloning vector pEX18Tc, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6369)
AUTHORS Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP.
TITLE A broad-host-range Flp-FRT recombination system for site-specific
excision of chromosomally-located DNA sequences: application for
isolation of unmarked Pseudomonas aeruginosa mutants
JOURNAL Gene 212 (1), 77-86 (1998)
PUBMED 9661666
REFERENCE 2 (bases 1 to 6369)
AUTHORS Hoang TT, Karkhoff-Schweizer RR, Kutchma AJ, Schweizer HP.
TITLE Direct Submission
JOURNAL Submitted (10-FEB-1998) Microbiology, Colorado State University,
Fort Collins, CO 80523, USA
REFERENCE 3 (bases 1 to 6369)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6369)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 6369)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene";
date: "1998"; volume: "212"; issue: "1"; pages: "77-86"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(10-FEB-1998) Microbiology, Colorado State University, Fort Collins,
CO 80523, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6369
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 2899..2921
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(212..1630)
/label=SacB
/note="secreted levansucrase that renders bacterial growth
sensitive to sucrose"
promoter complement(1631..2076)
/label=sacB promoter
/note="sacB promoter and control region"
oriT complement(2450..2559)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
terminator complement(3002..3029)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(3121..3207)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rRNA complement(3208..3327)
/product="Eschericia coli 5S ribosomal RNA"
primer_bind 3557..3573
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(3577..3633)
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(3643..3658)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(3666..3682)
/label=lac operator
/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(3690..3720)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(3735..3756)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4044..4632)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4908..6095)
/codon_start=1
/label=TcR
/note="tetracycline efflux protein"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
promoter complement(6170..6274)
/label=AmpR promoter