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。
pWB980 derived from pUB110 is an expression vector in Bacillus for its high copy number and high stability. It has a size of 3787 base pairs, which is considered manageable and does not impose significant metabolic burden on the host cells. One key feature of the pWB980 plasmid is its bacterial resistance attribute, specifically for kanamycin. This feature is extremely useful as it allows for the selection and identification of bacterial cells that have successfully taken up the plasmid. pWB980 harbors the P43 promoter whose transcription direction is consistent with majority of the pUB110-encoded genes. This high-level promoter is popular for driving gene expression in Bacillus subtilis. It is important to note that the pWB980 vector is not a shuttle vector. Users are recommended to treat bacterial cells carrying this plasmid with lysozyme to maximize the plasmid yield during extraction. pWB980-ori is a shuttle plasmid with an additional replication origin compared to pWB980.
- 载体名称:
- pWB980-ori
- 载体抗性:
- Kanamycin
- 载体长度:
- 4276 bp
- 载体类型:
- Bacillus Expression vector
- 拷贝数:
- High copy number
- 启动子:
- P43
- 感受态:
- DH10B
- 培养温度:
- 37℃
pWB980-ori 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。
pWB980-ori 载体序列
LOCUS Exported 4276 bp DNA circular SYN 14-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pWB980-ori
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4276)
AUTHORS A
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4276
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 309..1313
/codon_start=1
/gene="repB"
/product="RepB replication protein"
/label=repB
/note="from Enterococcus faecalis plasmid pAM-alpha-1"
/db_xref="GI:22652809"
/protein_id="AAN03827.1"
/translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
CDS 1473..2243
/codon_start=1
/product="aminoglycoside O-nucleotidyltransferase
ANT(4')-Ia [Staphylococcus aureus]"
/label=Kan
/translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHG
QFFSILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTT
FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD
SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
rep_origin complement(2373..2961)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 3727..3748
/label=pwb980-p43-F
promoter 3772..4063
/label=P43
CDS 4105..4191
/codon_start=1
/label=sacB signal peptide
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFA"