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:
- pUC-TTrepT
- 载体抗性:
- Ampicillin
- 载体长度:
- 5056 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Vieira J, Messing J.
pUC-TTrepT 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。
pUC-TTrepT 载体序列
LOCUS 40924_44928 5056 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pUC-TTrepT DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5056)
AUTHORS Vieira J, Messing J.
TITLE The pUC plasmids, an M13mp7-derived system for insertion mutagenesis
and sequencing with synthetic universal primers
JOURNAL Gene 19 (3), 259-268 (1982)
PUBMED 6295879
REFERENCE 2 (bases 1 to 5056)
AUTHORS Fujita A, Misumi Y, Koyama Y.
TITLE Two versatile shuttle vectors for Thermus thermophilus-Escherichia
coli containing multiple cloning sites, lacZalpha gene and kanamycin
or hygromycin resistance marker
JOURNAL Plasmid 67 (3), 272-275 (2012)
PUBMED 22252135
REFERENCE 3 (bases 1 to 5056)
AUTHORS Fujita A.
TITLE Direct Submission
JOURNAL Submitted (08-SEP-2011) Contact:Atsushi Fujita National Institute of
Advanced Science and Technology, Biomedical Research Institute;
1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan URL
:http://www.aist.go.jp/
REFERENCE 4 (bases 1 to 5056)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 5056)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene";
date: "1982"; volume: "19"; issue: "3"; pages: "259-268"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Plasmid";
date: "2012"; volume: "67"; issue: "3"; pages: "272-275"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(08-SEP-2011) Contact:Atsushi Fujita National Institute of Advanced
Science and Technology, Biomedical Research Institute; 1-1-1
Higashi, Tsukuba, Ibaraki 305-8566, Japan URL
:http://www.aist.go.jp/"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5056
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 106..127
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 142..172
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 180..196
/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)."
primer_bind 204..220
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 233..283
/note="polylinker
HindIII-NotI-SalI-NruI-AflII-EcoRV-Acc65I-EcoRI"
primer_bind complement(284..300)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 690..3062
/label=derived from pYK225 (derivative of pTT8)
/note="derived from pYK225 (derivative of pTT8)"
CDS complement(1197..2360)
/codon_start=1
/gene="repA"
/product="putative RepA protein"
/label=repA
/protein_id="BAL70402.1"
/translation="MVLRAYAALRGLSPEALRAHLLAPPLRPERAREAFQRPYLAHFAQ
TLPRYPYATDDPKEGVRIYKRENALKRVHVQVGHYPHAVLRLVVDVDLPWPQVEERIHA
LPPSLVLVNPRSGHFHAWYELDPIPLTPPPGREGSLKGALALLAEVEALLEAYYGADPG
YNGLLSRNPFLHPPEWTWGGGKRWSLRDLHRELRGLLPSGTRRRVDPGLASYGRNNALF
DRLRAEAYAHVALFRGVPGGEEAFRAWVEQRAHALNQSLFRDHPKGPLDPREVHHTAKS
VAKWTYRNYRGARVYPVSSTGRPDRSRLSPQARALIPPLQGQELQEAVREGGRRRGSRR
RQEAEEKLTEALKRLQARGERVTARALAREAGVKPHTASKWLKRMRE"
gene complement(1197..2360)
/gene="repA"
/label=repA
promoter 3150..3254
/label=AmpR promoter
CDS 3255..4112
/label=AmpR
/note="beta-lactamase"
rep_origin 4286..4874
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"