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:
- pN-PURO-PTP
- 载体抗性:
- Ampicillin
- 载体长度:
- 5650 bp
- 载体类型:
- Transfection vector
- 复制子:
- ori
- 载体来源:
- Schimanski B, Nguyen TN, Gunzl A.
pN-PURO-PTP 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。
pN-PURO-PTP 载体序列
LOCUS 40924_32963 5650 bp DNA circular SYN 18-DEC-2018
DEFINITION Transfection vector pN-PURO-PTP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5650)
AUTHORS Schimanski B, Nguyen TN, Gunzl A.
TITLE Highly efficient tandem affinity purification of trypanosome protein
complexes based on a novel epitope combination
JOURNAL Eukaryotic Cell 4 (11), 1942-1950 (2005)
PUBMED 16278461
REFERENCE 2 (bases 1 to 5650)
AUTHORS Schimanski B, Nguyen TN, Gunzl A.
TITLE Direct Submission
JOURNAL Submitted (17-AUG-2005) Department of Genetics and Developmental
Biology, University of Connecticut Health Center, 263 Farmington
Ave, Farmington, CT 06030, USA
REFERENCE 3 (bases 1 to 5650)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5650)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Eukaryotic
Cell"; date: "2005"; volume: "4"; issue: "11"; pages: "1942-1950"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-AUG-2005) Department of Genetics and Developmental Biology,
University of Connecticut Health Center, 263 Farmington Ave,
Farmington, CT 06030, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5650
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 21..125
/label=AmpR promoter
CDS 126..983
/label=AmpR
/note="beta-lactamase"
rep_origin 1157..1745
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2033..2054
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2069..2099
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2107..2123
/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 2131..2147
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2168..2186
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 2199..2204
/note="unique SacI restriction site for cloning the
resistance marker cassette"
misc_feature 2204..2702
/label=HSP70 genes 2 and 3 intergenic region
/note="HSP70 genes 2 and 3 intergenic region"
CDS 2703..3299
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 3315..4055
/label=beta-alpha tubulin intergenic region
/note="beta-alpha tubulin intergenic region"
misc_feature 4056..4061
/note="unique HindIII restriction site for cloning the
resistance marker cassette and the PTP cassette"
CDS 4526..4699
/label=ProtA
/note="IgG-binding unit of Staphylococcus aureus protein A"
CDS 4703..4873
/codon_start=1
/product="IgG-binding unit of Staphylococcus aureus protein
A"
/label=ProtA
/translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
EAKKLNDAQAPK"
CDS 4910..4930
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
misc_feature 4973..4998
/note="multiple cloning site containing unique restriction
sites for KpnI, ApaI, SnaBI and NotI for fusing the target
protein to the PTP sequence"
promoter complement(5007..5025)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5032..5048)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 5190..5645
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"