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。
pMD18-T Simple Vector is a specialized vector for efficient cloning of PCR products (TA Cloning). This vector is converted from pUC18 vector by inserting an EcoRV recognition site between XbaI and Sa1I recognition sites at the multiple cloning site of pUC18 vector, and then adding a “T” at the 3' end of both sides after enzymatic digestion with EcoR V. The pMD*18-T Vector is a special vector for highly efficient cloning of PCR products (TA Cloning). Because most of the thermophilic DNA polymerase PCR reaction has the characteristic of adding an “A” at the 3' end of the PCR product, the use of this product can greatly improve the efficiency of ligation and cloning of PCR products. Since this vector is constructed on the basis of pUC18 vector, it has the same function as pUC18 vec
- 载体名称:
- pMD18-T Simple
- 载体抗性:
- Ampicillin
- 载体长度:
- 2692 bp
- 载体类型:
- TA Cloning Vector
- 载体来源:
- Long JY, Chen YH, Xia JR.
- 拷贝数:
- High Copy
- 启动子:
- lac
- 5'引物:
- M13 fwd
- 3'引物:
- M13 rev
- 感受态:
- suitable for use in most common commercial host cells like TOP10, DH5α, TOP10F and JM109
pMD18-T Simple 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。
pMD18-T Simple 载体序列
LOCUS Exported 2692 bp DNA circular SYN 20-JAN-2025
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2692)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..2692
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(146..475)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITNSSSVPGDPLEISSTCRHASLALAVVLQRRDWENPGVTQL
NRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTI
CSDAA"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(471..487)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 495..511
/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(519..549)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 564..585
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(873..1461)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1632..2492)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2493..2597)
/gene="bla"
/label=AmpR promoter