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:
- pCtB1
- 载体抗性:
- Kanamycin
- 载体长度:
- 5457 bp
- 载体类型:
- Yeast dual vector
- 复制子:
- ori
- 宿主:
- Yeast
- 载体来源:
- Lee WY, Chung S-C., Song J-Y., Cho KM.
- 启动子:
- URA3
pCtB1 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。
pCtB1 载体序列
LOCUS 40924_13755 5457 bp DNA circular SYN 17-DEC-2018 DEFINITION Yeast dual vector pCtB1, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5457) AUTHORS Lee WY, Chung S-C., Song J-Y., Cho KM. TITLE Direct Submission JOURNAL Submitted (02-JUN-2014) BioMaterial Lab, Samsung Advanced Institute of Technology, 130 Samsung-ro, Suwon 443803, Republic of Korea REFERENCE 2 (bases 1 to 5457) TITLE Direct Submission REFERENCE 3 (bases 1 to 5457) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted (02-JUN-2014) BioMaterial Lab, Samsung Advanced Institute of Technology, 130 Samsung-ro, Suwon 443803, Republic of Korea" COMMENT SGRef: number: 2; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..5457 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin 134..1298 /label=2u ori /note="yeast 2u plasmid origin of replication" primer_bind 1356..1372 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 1379..1397 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature complement(1406..1513) /label=MCS /note="pBluescript multiple cloning site" CDS 1523..1549 /label=HA /note="HA (human influenza hemagglutinin) epitope tag" regulatory 1553..1799 /note="TPS1 terminator; interior" /regulatory_class="terminator" promoter 1821..2036 /label=URA3 promoter CDS 2037..2837 /label=URA3 /note="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" terminator 2841..3088 /label=CYC1 terminator /note="transcription terminator for CYC1" regulatory 3089..3301 /note="TPS1 terminator; lateral" /regulatory_class="terminator" promoter complement(3315..3333) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3354..3370) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3378..3394) /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(3402..3432) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3447..3468) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(3756..4344) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4517..5305) /label=NeoR/KanR /note="aminoglycoside phosphotransferase" promoter complement(5330..5434) /label=AmpR promoter