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:
- pP43NMK
- 载体抗性:
- Ampicillin
- 载体长度:
- 7683 bp
- 载体类型:
- Shuttle expression-secretion vector
- 复制子:
- ori
- 载体来源:
- Zhang XZ, Cui ZL, Hong Q, Li SP.
- 启动子:
- P43
- 感受态:
- DH5α
- 培养温度:
- 37℃
pP43NMK 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。
pP43NMK 载体序列
LOCUS pP43NMK 7683 bp DNA circular SYN 21-SEP-2024
DEFINITION Shuttle expression-secretion vector pP43NMK, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7683)
AUTHORS Zhang XZ, Cui ZL, Hong Q, Li SP.
TITLE High-level expression and secretion of methyl parathion hydrolase in
Bacillus subtilis WB800
JOURNAL Appl. Environ. Microbiol. 71 (7), 4101-4103 (2005)
PUBMED 16000826
REFERENCE 2 (bases 1 to 7683)
AUTHORS Zhang X-Z., Cui Z-L., Li S-P.
TITLE Direct Submission
JOURNAL Submitted (25-OCT-2005) Department of Microbiology, College of Life
Sciences, Nanjing Agricultural University, MOA Key Lab of
Microbiological Engineering of Agricultural Environment, #6 Tongwei
Road, Weigang, Nanjing, Jiangsu 210095, P. R. China
REFERENCE 3 (bases 1 to 7683)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7683)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 7683)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2005"; volume: "71"; issue: "7"; pages:
"4101-4103"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(25-OCT-2005) Department of Microbiology, College of Life Sciences,
Nanjing Agricultural University, MOA Key Lab of Microbiological
Engineering of Agricultural Environment, #6 Tongwei Road, Weigang,
Nanjing, Jiangsu 210095, P. R. China"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7683
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 814..1815
/label=repB
/note="RepB replication protein"
CDS 1987..2748
/codon_start=1
/gene="km"
/product="Km"
/label=km
/protein_id="ABD24446.1"
/translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
gene 1987..2748
/gene="km"
/label=km
CDS 2965..3369
/codon_start=1
/gene="ble"
/product="Ble"
/label=ble
/protein_id="ABD24447.1"
/translation="MRMLQSIPALPVGDIKKSIGFYCDKLGFTLVHHEDGFAVLMCNEV
RIHLWEASDEGWRSRSNDSPVCTGAESFIAGTASCRIEVEGIDELYQHIKPLGILHPNT
SLKDQWWDERDFAVIDPDNNLISFFQQIKS"
gene 2965..3217
/gene="ble"
/label=ble
promoter 4366..4421
/label=P43
regulatory 4370..4387
/note="-35 and -10 signal of promoter controlled by sigma
B"
/regulatory_class="minus_35_signal"
regulatory 4389..4394
/note="-35 and -10 signal of promoter controlled by sigma
A"
/regulatory_class="minus_35_signal"
regulatory 4403..4412
/note="-35 and -10 signal of promoter controlled by sigma
B"
/regulatory_class="minus_10_signal"
regulatory 4412..4417
/note="-35 and -10 signal of promoter controlled by sigma
A"
/regulatory_class="minus_10_signal"
regulatory 4444..4453
/gene="mpd"
/regulatory_class="ribosome_binding_site"
CDS 4463..5437
/codon_start=1
/gene="mpd"
/product="MPH"
/label=mpd
/note="methyl parathion hydrolase; includes NprB signal
peptide"
/protein_id="ABD24448.1"
/translation="MRNSTKTSLLLAGLCTAAQMVFVTHASAAAPQVRTSAPGYYRMLL
GDFEITALSDGTVALPVDKRLNQPAPKTQSALAKSFQKAPLETSVTGYLVNTGSKLVLV
DTGAAGLFGPTLGRLAANLKAAGYQPEQVDEIYITHMHPDHVGGLMVGEQLAFPNAVVR
ADQKEADFWLSQTNLDKAPDDESKGFFKGAMASLNPYVKAGKFKPFSGNTDLVPGIKAL
ASHGHTPGHTTYVVESQGQKLALLGDLILVAAVQFDDPSVTNQLDIDGKSAAVERKKAF
ADAAKGGYLIAASHLPFPGIGHIRAEGKGYRFVPVNYSVVNPK"
primer_bind complement(5462..5478)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5486..5502)
/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)."
promoter complement(5510..5540)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5555..5576)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5864..6452)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6626..7483)
/label=AmpR
/note="beta-lactamase"
promoter complement(7484..7588)
/label=AmpR promoter