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:
- pSH062
- 载体抗性:
- Ampicillin
- 载体长度:
- 10397 bp
- 载体类型:
- Cloning vector
- 复制子:
- p15A ori
- 载体来源:
- Hon S, Olson DG, Holwerda EK, Lanahan AA, Murphy SJL., Maloney MI, Zheng T, Papanek B, Guss AM, Lynd LR.
pSH062 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。
pSH062 载体序列
LOCUS V003257 10397 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V003257
VERSION V003257
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 10397)
AUTHORS Hon S, Olson DG, Holwerda EK, Lanahan AA, Murphy SJL., Maloney MI,
Zheng T, Papanek B, Guss AM, Lynd LR.
TITLE The ethanol pathway from Thermoanaerobacterium saccharolyticum
improves ethanol production in Clostridium thermocellum
JOURNAL Metab. Eng. 42, 175-184 (2017)
PUBMED 28663138
REFERENCE 2 (bases 1 to 10397)
AUTHORS Hon S, Olson DG, Holwerda EK, Lanahan AA, Murphy SJL., Maloney MI,
Zheng T, Papanek BA, Guss AM, Lynd LR.
TITLE Direct Submission
JOURNAL Submitted (27-MAR-2017) Thayer School of Engineering, Dartmouth
College, 14 Engineering Drive, Hanover, NH 03755, USA
REFERENCE 3 (bases 1 to 10397)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10397)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
42, 175-184 (2017)"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(27-MAR-2017) Thayer School of Engineering, Dartmouth College, 14
Engineering Drive, Hanover, NH 03755, USA"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10397
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 1..92
/label="cat promoter"
/note="cat promoter"
/regulatory_class="promoter"
CDS 93..740
/gene="cat"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from
Staphylococcus aureus. Accession#: P00485"
CDS complement(824..1681)
/label="AmpR"
/note="beta-lactamase"
promoter complement(1682..1773)
/label="AmpR promoter"
rep_origin 2033..2578
/direction=RIGHT
/label="p15A ori"
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
CDS 3049..4050
/label="repB"
/note="RepB replication protein"
regulatory 4099..4307
/label="Clo1313_2638 promoter"
/note="Clo1313_2638 promoter"
/regulatory_class="promoter"
CDS 4308..6887
/codon_start=1
/gene="adhE"
/product="AdhE"
/label="adhE"
/note="bifunctional alcohol and aldehyde dehydrogenase"
/protein_id="ASK86031.1"
/translation="MATTKTELDVQKQIDLLVSRAQEAQKKFMSYTQEQIDAIVKAMAL
AGVDKHVELAKMAYEETKMGVYEDKITKNLFATEYVYHDIKNEKTVGIINENIEENYME
VAEPIGVIAGVTPVTNPTSTTMFKCLISIKTRNPIIFSFHPKAIKCSIAAAKVMYEAAL
KAGAPEGCIGWIETPSIEATQLLMTHPGVSLILATGGAGMVKAAYSSGKPALGVGPGNV
PCYIEKSANIKRAVSDLILSKTFDNGVICASEQAVIIDEEIADEVKKLMKEYGCYFLNK
DEIKKLEKFAIDEQSCAMSPAVVGQPAAKIAEMAGFKVPEGTKILVAEYEGVGPKYPLS
REKLSPILACYTVKDYNEGIKKCEEMTEFGGLGHSAVIHSENQNVINEFARRVRTGRLI
VNSPSSQGAIGDIYNTNTPSLTLGCGSMGRNSTTDNVSVKNLLNIKRVVIRKDRMKWFK
IPPKIYFESGSLQYLCKVKRKKAFIVTDPFMVKLGFVDKVTYQLDKANIEYEIFSEVEP
DPSVDTVMNGVKIMNSYNPDLIIAVGDGSAIDAAKGMWLFYEYPDTEFETLRLKFADIR
KRAFKFPELGKKALFIAIPTTSGTGSEVTAFAVITDKKRNIKYPLADYELTPDIAIIDP
DLTKTVPPSVTADTGMDVLTHAIEAYVSVMASDYTDALAEKAIKIVFEYLPRAYKNGND
EEAREKMHNASCMAGMAFTNAFLGINHSMAHILGGKFHIPHGRANAILLPYVIRYNAEK
PTKFVAFPQYEYPKAAERYAEIAKFLGLPASTVEEGVESLIEAIKNLMKELNIPLTLKD
AGINKEQFEKEIEEMSDIAFNDQCTGTNPRMPLTKEIAEIYRKAYGA"
gene 4308..6887
/gene="adhE"
/label="adhE"
CDS 6905..7759
/codon_start=1
/gene="nfnA"
/product="NfnA"
/label="nfnA"
/note="NADH-dependent reduced ferredoxin:NADP+
oxidoreductase"
/protein_id="ASK86033.1"
/translation="MNEILEKKQLNPTVKMMVINAPLMAKKAKPGQFVIVRVDEKGERI
PLTIADYDRNKGTITIIFQEVGMSTKKLGTLNVGDRLHDFVGPLGKPVEFSKDTKRVLA
IGGGVGVAPLYPKVKMLNEMKVPVDSIIGGRSAEYVILEDEMKKVSENLYITTDDGTKG
RKGFVTDVLKELIEKDNKYDEVIAIGPLIMMKMVCNITKEYNIPTMVSMNPIMIDGTGM
CGGCRVTVGGETKFACVDGPAFDGLKVDFDEAMRRQNMYKDMERKVLENYEHECKLGGI
LNG"
gene 6905..7759
/gene="nfnA"
/label="nfnA"
CDS 7752..9143
/codon_start=1
/gene="nfnB"
/product="NfnB"
/label="nfnB"
/note="NADH-dependent reduced ferredoxin:NADP+
oxidoreductase"
/protein_id="ASK86034.1"
/translation="MANMSLKKVPMPEQEPDQRNKNFKEVALGYEENMAVEEAERCIQC
KNQPCVEGCPVHVKIPEFIKLIANRDFEGAYQKIKETNNLPAICGRVCPQESQCESVCT
RGKKGEPVAIGRLERFTADWHMKNNEDKIEKPETNGRKVAVIGSGPAGLSCAGDLAKMG
YDTTIFEAFHTPGGVLMYGIPEFRLPKEIVQKEIDSLKKLGVKIETNMVIGKILTIDDL
FDMGYEAVFIGTGAGLPKFMNIPGENLNGVYSANEFLTRINLMKAYDFPNSPTPVKVGK
KVAVVGGGNVAMDAARSAKRMGAEEVYIVYRRSEEEMPARLEEIHHAKEEGIIFKLLTN
PVRIIGDESGSVKGIECVNMVLGDVDESGRRRPVEEKGSEHVIDVDTVIIAIGQSPNPL
ITSTTEGLEKQRWGGIIVNEETLETSRRGVFAGGDAVTGAATVILAMGAGKKAAASIHK
YLSEK"
gene 7752..9143
/gene="nfnB"
/label="nfnB"
CDS 9161..10360
/codon_start=1
/gene="adhA"
/product="AdhA"
/label="adhA"
/note="NADPH-dependent alcohol dehydrogenase"
/protein_id="ASK86032.1"
/translation="MWETKVNPSKIFELRCKNTTYFGVGSIHKIKDILENLKINGINNV
IFITGKGSYKTSGAWDVVRPVLEELDLKYSLYDKVGPNPTVDMIDEAAKIGRESGAKAV
IGIGGGSPIDTAKSVAVLLKYTDKNARELYKQKFIPDDAVPIIAINLTHGTGTEVDRFA
VATIPEKNYKPAIAYDCLYPMFAIDDPSLMTKLDKKQTIAVTVDALNHITEAATTLVAS
PYSILTAKETVRLIVRYLPAAVNDPLNIVARYYLLYASALAGISFDNGLLHLTHALEHP
LSAVKPEIAHGLGLGAILPAVIKAIYPATAEVLADVYSPIVPGLKGLPVEAEYVAEKVQ
EWLFSVGCIQKLSDFGFTKDDIPNLVKLAKTTPSLDGLLSIAPVEATESVIEKIYLKSL
"
gene 9161..10360
/gene="adhA"
/label="adhA"