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学术研究使用，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。
Note:
- 载体名称:
- pAH-mini-Mu(LER)-YK
- 载体抗性:
- Kanamycin
- 载体长度:
- 7289 bp
- 载体类型:
- Cloning vector
- 复制子:
- R6K γ ori
- 载体来源:
- Gorshkova NV, Lobanova JS, Tokmakova IL, Smirnov SV, Akhverdyan VZ, Krylov AA, Mashko SV.
- 启动子:
- tetR/tetAs
pAH-mini-Mu(LER)-YK 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。
pAH-mini-Mu(LER)-YK 载体序列
LOCUS 40924_4150 7289 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pAH-mini-Mu(LER)-YK, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7289)
AUTHORS Gorshkova NV, Lobanova JS, Tokmakova IL, Smirnov SV, Akhverdyan VZ,
Krylov AA, Mashko SV.
TITLE Mu-driven transposition of recombinant mini-Mu unit DNA in the
Corynebacterium glutamicum chromosome
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7289)
AUTHORS Gorshkova NV, Lobanova JS, Tokmakova IL, Smirnov SV, Akhverdyan VZ,
Krylov AA, Mashko SV.
TITLE Direct Submission
JOURNAL Submitted (26-SEP-2017) Ajinomoto-Genetika Research Institute, 1st
Dorozhny pr. 1-1, Moscow 117545, Russian Federation
REFERENCE 3 (bases 1 to 7289)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7289)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-SEP-2017) Ajinomoto-Genetika Research Institute, 1st Dorozhny
pr. 1-1, Moscow 117545, Russian Federation"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7289
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_recomb complement(58..174)
/label=Mu-R
/note="Mu-R"
regulatory complement(179..276)
/label=deoT
/note="deoT"
/regulatory_class="terminator"
protein_bind complement(282..315)
/label=lox71
/note="Left element (LE) mutant of loxP (Araki et al.,
2010). Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
protein_bind 361..394
/label=FRT (minimal)
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
CDS complement(588..1379)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
protein_bind 1739..1786
/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)."
regulatory complement(1837..2204)
/label=Mu phage E-element
/note="Mu phage E-element"
/regulatory_class="enhancer"
CDS complement(2216..3052)
/codon_start=1
/product="streptomycin resistance protein subunit B"
/label=streptomycin resistance protein subunit B
/note="strB"
/protein_id="ATO98411.1"
/translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
CDS complement(3052..3855)
/codon_start=1
/product="streptomycin resistance protein subunit A"
/label=streptomycin resistance protein subunit A
/note="strA"
/protein_id="ATO98412.1"
/translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
protein_bind complement(3925..3958)
/label=lox66
/note="Right element (RE) mutant of loxP (Araki et al.,
2010). Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS complement(3962..4675)
/label=Citrine
/note="enhanced variant of YFP (Heikal et al., 2001)"
regulatory complement(4716..4837)
/label=P17
/note="P17"
/regulatory_class="promoter"
primer_bind complement(4909..4925)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
terminator 5128..5187
/label=his operon terminator
/note="This putative transcriptin terminator from the E.
coli his operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined."
misc_recomb complement(5199..5416)
/label=Mu-L
/note="Mu-L"
rep_origin complement(5546..5934)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(6024..7226)
/label=TcR
/note="tetracycline efflux protein"
protein_bind complement(7235..7253)
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"