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:
- pP02218-nat-cel8A
- 载体抗性:
- Ampicillin
- 载体长度:
- 6904 bp
- 载体类型:
- Shuttle vector
- 复制子:
- ori
- 载体来源:
- Heinze S, Zimmermann K, Ludwig C, Heinzlmeir S, Schwarz WH, Zverlov VV, Liebl W, Kornberger P.
pP02218-nat-cel8A 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。
pP02218-nat-cel8A 载体序列
LOCUS V004191 6904 bp DNA circular SYN 18-DEC-2018 DEFINITION Exported. ACCESSION V004191 VERSION V004191 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 6904) AUTHORS Heinze S, Zimmermann K, Ludwig C, Heinzlmeir S, Schwarz WH, Zverlov VV, Liebl W, Kornberger P. TITLE Efficient secretory heterologous enzyme production in the novel host Paenibacillus polymyxa by usage of its promoter sequences JOURNAL Unpublished REFERENCE 2 (bases 1 to 6904) AUTHORS Heinze S, Zimmermann K, Ludwig C, Heinzlmeir S, Schwarz WH, Zverlov VV, Liebl W, Kornberger P. TITLE Direct Submission JOURNAL Submitted (09-MAY-2018) Department for Microbiology, Technical University of Munich, Emil-Ramann-Str. 4, Freising, Bavaria 85354, Germany REFERENCE 3 (bases 1 to 6904) TITLE Direct Submission REFERENCE 4 (bases 1 to 6904) AUTHORS . TITLE Direct Submission COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## SGRef: number: 1; type: "Journal Article"; journalName: "Unpublished" SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (09-MAY-2018) Department for Microbiology, Technical University of Munich, Emil-Ramann-Str. 4, Freising, Bavaria 85354, Germany" SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6904 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin complement(79..667) /direction=LEFT /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(841..1698) /label="AmpR" /note="beta-lactamase" promoter complement(1699..1803) /label="AmpR promoter" CDS complement(1832..2116) /codon_start=1 /gene="traJ" /product="TraJ" /label="traJ" /protein_id="AXF54530.1" /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGQQKAHYVNHHPNQVFWGRGAVKH" gene complement(1832..2116) /gene="traJ" /label="traJ" oriT complement(2149..2258) /direction=LEFT /label="oriT" /note="incP origin of transfer" CDS complement(2515..3276) /codon_start=1 /gene="kanR" /product="KanR" /label="kanR" /note="Kanamycin Resistance" /protein_id="AXF54531.1" /translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK LLESLENFWNGIQEWTERHGYIVDVSKRIPF" gene complement(2515..3276) /gene="kanR" /label="kanR" CDS complement(3448..4449) /label="repB" /note="RepB replication protein" regulatory 4866..5277 /label="P02218 promoter from P. polymyxa DSM292" /note="P02218 promoter from P. polymyxa DSM292" /regulatory_class="promoter" CDS 5308..6717 /gene="celA" /label="Endoglucanase A" /note="Endoglucanase A from Acetivibrio thermocellus (strain ATCC 27405 / DSM 1237 / JCM 9322 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372). Accession#: A3DC29" CDS 6724..6741 /label="6xHis" /note="6xHis affinity tag"