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:
- pCotZ-CL-thr
- 载体抗性:
- Ampicillin
- 载体长度:
- 7655 bp
- 载体类型:
- Cloning vector
- 复制子:
- ori
- 载体来源:
- Iwanicki A, Hinc K, Stasilojc M, Piatek I, Grela A, Lega T, Obuchowski M.
pCotZ-CL-thr 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。
pCotZ-CL-thr 载体序列
LOCUS V008272 7655 bp DNA circular SYN 17-DEC-2018
DEFINITION Exported.
ACCESSION V008272
VERSION V008272
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7655)
AUTHORS Iwanicki A, Hinc K, Stasilojc M, Piatek I, Grela A, Lega T,
Obuchowski M.
TITLE Vector system for spore sufrace display
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7655)
AUTHORS Iwanicki A, Hinc K, Stasilojc M, Piatek I, Grela A, Lega T,
Obuchowski M.
TITLE Direct Submission
JOURNAL Submitted (25-NOV-2013) Laboratory of Molecular Bacteriology,
Medical University of Gdansk, Debinki 1, Gdansk 80-211, Poland
REFERENCE 3 (bases 1 to 7655)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7655)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Assembly Method :: Vector NTI Advanced v. 10
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(25-NOV-2013) Laboratory of Molecular Bacteriology, Medical
University of Gdansk, Debinki 1, Gdansk 80-211, Poland"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7655
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 247..879
/codon_start=1
/gene="thrC"
/product="ThrC"
/label="thrC"
/note="N terminal part of thrC"
/protein_id="AHG54660.1"
/translation="MWKGLIHQYKEFLPVTDQTPALTLHEGNTPLIHLPKLSEQLGIEL
HVKTEGVNPTGSFKDRGMVMAVAKAKEEGNDTIMCASTGNTSAAAAAYAARANMKCIVI
IPNGKIAFGKLAQAVMYGAEIIAIDGNFDDALKIVRSICEKSPIALVNSVNPYRIEGQK
TAAFEIVHNGFPYAKYGQTWPARLLLFLTPDQLVMVATGAQLELRRY"
gene 247..879
/gene="thrC"
/label="thrC"
gene complement(1124..2316)
/gene="cotZ"
/label="cotZ"
misc_feature complement(1124..1149)
/gene="cotZ"
/label="multiple cloning site"
/note="multiple cloning site"
CDS complement(1135..1626)
/codon_start=1
/gene="cotZ"
/product="CotZ"
/label="cotZ"
/note="spore coat protein"
/protein_id="AHG54656.1"
/translation="MSQKTSSCVREAVENIEDLQNAVEEDCPTGCHSKLLSVSHSLGDT
VPFAIFTSKSTPLVAFGNVGELDNGPCFNTVFFRVERVHGSCATLSLLIAFDEHKHILD
FTDKDTVCEVFRLEKTNYCIEVDLDCFCAINCLNPRLINRTHHHGGGEAAAKGGGGSGR
"
regulatory complement(1633..1638)
/gene="cotZ"
/regulatory_class="ribosome_binding_site"
CDS complement(1754..2293)
/codon_start=1
/gene="cotY"
/product="CotY"
/label="cotY"
/note="spore coat protein"
/protein_id="AHG54655.1"
/translation="MLLRTKVKYPIKKELKSMSCGKTHGRHENCVCDAVEKILAEQEAV
EEQCPTGCYTNLLNPTIAGKDTIPFLVFDKKGGLFSTFGNVGGFVDDMQCFESIFFRVE
KLCDCCATLSILRPVDVKGDTLSVCHPCDPDFFGLEKTDFCIEVDLGCFCAIQCLSPEL
VDRTSPHKDKKHHHNG"
gene complement(1754..2293)
/gene="cotY"
/label="cotY"
regulatory complement(2288..2316)
/gene="cotZ"
/regulatory_class="promoter"
CDS 2536..3288
/codon_start=1
/gene="trpC"
/product="TrpC"
/label="trpC"
/protein_id="AHG54661.1"
/translation="MLEKIIKQKKEEVKTLVLPVEQPFEKRSFKEAPASPNRFIGLIAE
VKKASPSKGLIKEDFVPVQIAKDYEAAKADAISVLTDTPFFQGENSYLSDVKRAVSIPV
LRKDFIIDSLQVEESRRIGADAILLIGEVLDPLHLHELYLEAGEKGMDVLVEVHDASTL
EQILKVFTPDILGVNNRNLKTFETSVKQTEQIASLVPKESLLVSESGIGSLEHLTFVNE
HGARAVLIGESLMRQTSQRKAIHALFRE"
gene 2536..3288
/gene="trpC"
/label="trpC"
CDS 3718..4146
/codon_start=1
/gene="thrC"
/product="ThrC"
/label="thrC"
/note="C-terminal part of thrC"
/protein_id="AHG54659.1"
/translation="MRGFEAEGAAAIVRNEVIENPETIATAIRIGNPASWDKAVKAAEE
SNGKIDEVTDDEILHAYQLIARVEGVFAEPGSCASIAGVLKQVKSGEIPKGSKVVAVLT
GNGLKDPNTAVDISEIKPVTLPTDEDSILEYVKGAARV"
gene 3718..4146
/gene="thrC"
/label="thrC"
CDS 4143..4553
/codon_start=1
/gene="thrB"
/product="ThrB"
/label="thrB"
/note="N terminal part of thrB"
/protein_id="AHG54658.1"
/translation="MNEADMLFSVTVPGSTANLGPGFDSVGMALSRYLKLTVFESDKWS
FEAETETVAGINSSSFNFLTSSCIGTISRSSNPSSFTSIRFNQHSKNLTGHRQFFEQCA
NYQLFLQFIQGTGEPACNPSCSIMRNDPHPVS"
gene 4143..4553
/gene="thrB"
/label="thrB"
CDS 4797..5576
/gene="ant1"
/label="Spectinomycin 9-adenylyltransferase"
/note="Spectinomycin 9-adenylyltransferase from
Staphylococcus aureus (strain N315). Accession#: P0A0D1"
rep_origin complement(5826..6414)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6588..7445)
/label="AmpR"
/note="beta-lactamase"
promoter complement(7446..7550)
/label="AmpR promoter"