Note:
基本信息
- 载体名称:
- pUC18T-mini-Tn7T-lux-Tp
- 载体抗性:
- Ampicillin
- 载体长度:
- 11739 bp
- 载体类型:
- Reporter vector
- 复制子:
- ori
- 载体来源:
- Damron FH, McKenney ES, Barbier M, Liechti GW, Schweizer HP, Goldberg JB.
- 启动子:
- Pc
- 感受态:
- DH10B
- 培养温度:
- 37℃
产品信息
质粒编号:V002410
质粒名称:pUC18T-mini-Tn7T-lux-Tp
规格:5 μg (冻干粉)
下载资源
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。
确保质粒的关键元件正确，但是我们并不能保证实验效果。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。
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。
pUC18T-mini-Tn7T-lux-Tp 质粒 (编号: V002410)序列
LOCUS Exported 11739 bp DNA circular SYN 21-JUL-2025
DEFINITION Reporter vector pUC18T-mini-Tn7T-lux-Tp, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11739)
AUTHORS Damron FH, McKenney ES, Barbier M, Liechti GW, Schweizer HP,
Goldberg JB.
TITLE Construction of conjugatable Mini-Tn7 Vectors for Bioluminescent
Detection and Single Copy Promoter lux Reporter Analysis in
Gram-negative Bacteria
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 11739)
AUTHORS Damron FH, McKenney ES, Barbier M, Liechti GW, Schweizer HP,
Goldberg JB.
TITLE Direct Submission
JOURNAL Submitted (30-MAR-2013) Microbiology, Immunology, and Cancer
Biology, University of Virginia, PO Box 800734, Charlottesville, VA
22908, USA
REFERENCE 3 (bases 1 to 11739)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11739)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 11739)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(30-MAR-2013) Microbiology, Immunology, and Cancer Biology,
University of Virginia, PO Box 800734, Charlottesville, VA 22908,
USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..11739
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 349..365
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 393..487
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
terminator 590..676
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
protein_bind 722..769
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(834..1067)
/label=TpR
/note="E. coli plasmid-associated dihydrofolate reductase"
regulatory 1263..1304
/label=promoter 2
/note="promoter 2"
/regulatory_class="promoter"
promoter complement(1393..1421)
/label=Pc promoter
/note="class 1 integron promoter"
misc_feature 1473..1520
/label=FRT
/note="FRT"
protein_bind 1473..1520
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
primer_bind 1473..1495
/label=Plux seq F
/note="Plux seq F"
promoter 1616..1644
/label=Pc promoter
/note="class 1 integron promoter"
primer_bind 2041..2063
/label=Plux seq R
/note="Plux seq R"
CDS 2491..3930
/label=LuxC
/note="LuxC fatty acid reductase"
CDS 3946..4866
/codon_start=1
/label=LuxD
/note="LuxD acyltransferase"
/translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
FEHLTIATVNERRMRIEIENQAISLS"
CDS 4918..5997
/label=LuxA
/note="LuxA luciferase subunit"
CDS 6015..6995
/codon_start=1
/label=LuxB
/note="LuxB luciferase subunit"
/translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILG
FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
CDS 7177..8286
/label=LuxE
/note="LuxE"
mobile_element complement(9020..9185)
/label=Tn7L
/note="mini-Tn7 element (left end of the Tn7 transposon)"
oriT complement(9344..9452)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
rep_origin complement(9684..10272)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10446..11303)
/label=AmpR
/note="beta-lactamase"
promoter complement(11304..11408)
/label=AmpR promoter