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:
- pSV2neo
- 载体抗性:
- Ampicillin
- 载体长度:
- 5729 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Southern PJ, Berg P.
- 启动子:
- SV40
pSV2neo 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。
pSV2neo 载体序列
LOCUS 40924_41729 5729 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pSV2neo, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5729)
AUTHORS Southern PJ, Berg P.
TITLE Transformation of mammalian cells to antibiotic resistance with a
bacterial gene under control of the SV40 early region promoter
JOURNAL J. Mol. Appl. Genet. 1 (4), 327-341 (1982)
PUBMED 6286831
REFERENCE 2 (bases 1 to 5729)
AUTHORS Kitts PA.
TITLE CLONTECH Vectors On Disc version 1.3
JOURNAL Unpublished
REFERENCE 3 (bases 1 to 5729)
AUTHORS Kitts PA.
TITLE Direct Submission
JOURNAL Submitted (07-OCT-1993) Paul A. Kitts, CLONTECH Laboratories, Inc.,
1020 East Meadow Circle, Palo Alto, CA 94303, USA
REFERENCE 4 (bases 1 to 5729)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 5729)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Mol.
Appl. Genet."; date: "1982"; volume: "1"; issue: "4"; pages:
"327-341"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
(07-OCT-1993) Paul A. Kitts, CLONTECH Laboratories, Inc., 1020 East
Meadow Circle, Palo Alto, CA 94303, USA"
COMMENT SGRef: number: 4; type: "Journal Article"
COMMENT This vector can be obtained from CLONTECH Laboratories, Inc., 1020
East Meadow Circle, Palo Alto, CA 94303, USA. To place an order call
(415) 424-8222 or (800) 662-2566, extension 1. International
customers, please contact your local distributor. For technical
information, call (415) 424- 8222 or (800) 662-2566, extension 3.
This sequence has been compiled from information in the sequence
databases, published literature and other sources, together with
partial sequences obtained by CLONTECH; this vector has not been
completely sequenced. If you suspect there is an error in this
sequence, please contact CLONTECH's Technical Service Department at
(415) 424-8222 or (800) 662-2566, extension 3 or E-mail
TECH@CLONTECH.COM.
FEATURES Location/Qualifiers
source 1..5729
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal complement(752..886)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
CDS complement(1311..1331)
/codon_start=1
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
intron complement(1461..1526)
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS complement(1950..2741)
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(3098..3427)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
misc_feature 3578..3718
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(3904..4492)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4666..5523)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5524..5628)
/label=AmpR promoter