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:
- pTEX5501ts
- 载体抗性:
- Chloramphenicol
- 载体长度:
- 6279 bp
- 载体类型:
- Shuttle vector
- 复制子:
- ori
- 载体来源:
- Nallapareddy SR, Singh KV, Murray BE.
pTEX5501ts 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。
pTEX5501ts 载体序列
LOCUS V002746 6279 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V002746
VERSION V002746
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6279)
AUTHORS Nallapareddy SR, Singh KV, Murray BE.
TITLE Construction of Improved Temperature-Sensitive and Mobilizable
Vectors and Their Use for Constructing Mutations in the
Adhesin-Encoding acm Gene of Poorly Transformable Clinical
Enterococcus faecium Strains
JOURNAL Appl. Environ. Microbiol. 72 (1), 334-345 (2006)
PUBMED 16391062
REFERENCE 2 (bases 1 to 6279)
AUTHORS Nallapareddy SR, Singh KV, Murray BE.
TITLE Direct Submission
JOURNAL Submitted (18-SEP-2005) Infectious Diseases, Univ. of Texas Med.
School at Houston, 6431 Fannin St, MSB 2.112, Houston, TX 77030, USA
REFERENCE 3 (bases 1 to 6279)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6279)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2006"; volume: "72"; issue: "1"; pages:
"334-345"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(18-SEP-2005) Infectious Diseases, Univ. of Texas Med. School at
Houston, 6431 Fannin St, MSB 2.112, Houston, TX 77030, USA"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6279
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..61
/note="multiple cloning site I; MCS-I"
CDS 195..842
/gene="cat"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
misc_feature 1007..1076
/note="multiple cloning site II; MCS-II"
CDS complement(1479..1490)
/label="Factor Xa site"
/note="Factor Xa recognition and cleavage site"
CDS 1624..1833
/codon_start=1
/gene="orfB"
/product="OrfB"
/label="orfB"
/note="portion of pWV01 based temperature-sensitive
gram-positive replicon derived from pTV1-OK"
/protein_id="ABB20801.1"
/translation="MGGKEANFASVLRPPIKCRVPIFVPKTLYPNWLKGLRGFSIANES
PTFSPTFFINLYLSSFIVVFMITK"
gene 1624..1833
/gene="orfB"
/label="orfB"
CDS 1874..2035
/codon_start=1
/product="OrfC"
/label="OrfC"
/note="portion of pWV01 based temperature-sensitive
gram-positive replicon derived from pTV1-OK"
/protein_id="ABB20802.1"
/translation="MVISESKKRVMISLTKEQDKKLTDMAKQKGFSKSAVAALAIEEYA
RKESEQKK"
CDS 2102..2800
/codon_start=1
/gene="repA(ts)"
/product="RepA(ts)"
/label="repA(ts)"
/note="portion of pWV01 based temperature-sensitive
gram-positive replicon derived from pTV1-OK"
/protein_id="ABB20803.1"
/translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDLYITLDESQKRELKNLLLDIV
DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
VLDAETGEIK"
gene 2102..2800
/gene="repA(ts)"
/label="repA(ts)"
CDS 2797..2946
/codon_start=1
/gene="orfD"
/product="OrfD"
/label="orfD"
/note="portion of pWV01 based temperature-sensitive
gram-positive replicon derived from pTV1-OK"
/protein_id="ABB20804.1"
/translation="MTNKEKELFAENEELKKEIKDLKERIERYREMEVELSTTIDLLRG
GIIE"
gene 2797..2946
/gene="orfD"
/label="orfD"
CDS 3173..4078
/codon_start=1
/gene="aph(2'')-Id"
/product="APH(2'')-Id"
/label="aph(2'')-Id"
/note="aminoglycoside modifying enzyme; derived from
Enterococcus casseliflavus in GenBank Accession Number
AF016483"
/protein_id="ABB20805.1"
/translation="MRTYTFDQVEKAIEQLYPDFTINTIEISGEGNDCIAYEINRDFIF
KFPKHSRGSTNLFNEVNILKRIHNKLPLPIPEVVFTGMPSETYQMSFAGFTKIKGVPLT
PLLLNNLPKQSQNQAAKDLARFLSELHSINISGFKSNLVLDFREKINEDNKKIKKLLSR
ELKGPQMKIVDDFYRDILENEIYFKYYPCLIHNDFSSDHILFDTEKNTICGIIDFGDAA
ISDPDNDFISLMEDDEEYGMEFVSKILNHYKHKDIPTVLEKYRMKEKYWSFEKIIYGKE
YGYMDWYEEGLNEIRSIKIK"
gene 3173..4078
/gene="aph(2'')-Id"
/label="aph(2'')-Id"
oriT 4172..4838
/note="oriT of Tn916 derived from pHS1; facilitates
conjugative transfer from the transformable E. faecalis
strain JH2Sm::Tn916 to E. faecium"
rep_origin complement(5036..5624)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"