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:
- pLX-UCBSV
- 载体抗性:
- Kanamycin
- 载体长度:
- 13212 bp
- 载体类型:
- Cloning vector
- 复制子:
- pBBR1 oriV
- 宿主:
- Plants
- 载体来源:
- Pasin F, Bedoya LC, Bernabe-Orts JM, Gallo A, Simon-Mateo C, Orzaez D, Garcia JA.
- 启动子:
- CaMV 35S (enhanced)
pLX-UCBSV 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。
pLX-UCBSV 载体序列
LOCUS 40924_29436 13212 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pLX-UCBSV, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13212)
AUTHORS Pasin F, Bedoya LC, Bernabe-Orts JM, Gallo A, Simon-Mateo C, Orzaez
D, Garcia JA.
TITLE Multiple T-DNA Delivery to Plants Using Novel Mini Binary Vectors
with Compatible Replication Origins
JOURNAL ACS Synth Biol (2017) In press
PUBMED 28657330
REFERENCE 2 (bases 1 to 13212)
AUTHORS Pasin F, Garcia JA.
TITLE Direct Submission
JOURNAL Submitted (26-MAR-2017) Department of Plant Molecular Genetics,
Centro Nacional de Biotecnologia (CNB-CSIC), Darwin 3, Madrid,
Madrid 28049, Spain
REFERENCE 3 (bases 1 to 13212)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 13212)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol (2017) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-MAR-2017) Department of Plant Molecular Genetics, Centro
Nacional de Biotecnologia (CNB-CSIC), Darwin 3, Madrid, Madrid
28049, Spain"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13212
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 1..24
/label=assembly linker
/note="assembly linker"
regulatory 25..72
/note="artificial; tested only in sense orientation"
/regulatory_class="terminator"
misc_feature 133..157
/label=LB T-DNA repeat
/note="left border repeat from octopine Ach5 T-DNA"
misc_feature 184..208
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
promoter 514..1183
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
misc_feature 1183..10275
/note="DSMZ: PV-0912; Ugandan cassava brown streak virus
isolate Ke_125"
CDS 1317..10025
/codon_start=1
/product="polyprotein"
/label=polyprotein
/protein_id="ASG92166.1"
/translation="MSTIQLFKTIRFGSFEPVKLDEGNNIIEKVPVDLLAGNDGSGPEE
QSEQKYHRKESGESWRKVTDLYSVIGNSVYCRSYVAMKNFLNDTKWGGLFKNKKGQELK
AAARLRRATSYGFMYDPVACAFECPVCRTKATALEAFTSDCDHCFEIKHIDDDRIVQTE
TKFYPINPIELDVEDNLVEAASLEWLKGDVEERVVDRVLLLEDKEIHVTKKALVKRRAE
TKLVANAADLTKKLTEICCESGIPIIDIDNSKRKAIPMVKLKHIFGKVECDDMFEEDRY
FLEHSNAGKIFRSCEKITYRMIRPGWSGAIILRDNVQSEDHDKFDFINDICVVQGKNLI
SYKIENAMRVKTETELDLIDLYSFNLSWAKSKDKFIKHFESDTTQLVRSCCTPSTLWLY
ARKARFYKFVDHMILKGSPIIDILVKMEYVGKHLDMFNSVEDVCTEYSHFMKELISETV
NDKSDPDVLRVRNLIRAHFESVLEYNKYELIDRIIEKKTQLEAQEIMSRELIKHQYGEL
FSWRERLCLKLGIGSSNLYTYWVEREEKQSEKSNVISCIISKPGIEMLINWVSEVCKSK
YHSLVNCVDSGILFLWSRIVHLARVSVYGYWNLWFRQAMCVLFIFLVSNFSGKIVSYLK
KLIVSEKKLAIKNEEGFVEVQGRKEESFVLKWCAAATLFLSFLNYDWAVGCVSAIGKMK
TIFSALGPDFIEKQDGDDDLKFTTFEVEIPGDSRSSSAQTFGEWIEHCIKFNLVSIEPT
TSGPMLTLERGKANELADQINCLNATDIRVHGGVGTGKSTALPYELIRYGAVLVCVPTR
VLANALHESFMSLFGFDVSLAYRGRVRTGSKPITIMTYGYALNHFHHNPRNLAQFQFII
MDEVHTFPVHLNPLFSLLRELSPDKKIIKTSATHVGHSVDLSTNHKVDIHTLEIMDVKK
WAELQGTSVFGDVTKEPGNVLVFVASYRDVDVCSDKLKDKGFPVIKVDGRNFRKNTEVQ
KMVDGMQGEVKFIVATNIIENGVTLDVDVVVDFGERISPNLCSEDRCILMQRQRISQAE
RKQRFGRVGRMKRGSVYKFGRETLPDSMRNRVGSTESALICFAYGLKPVVDDVDIGSVR
SVTQRQALTASMFEANYIFTAHLVDKQGFMPRPVFELMKSLLLHTDAVGVSSTYLATNM
SGWRRLKEYIRIDDNSRHVQEVQIPWYCSDMSDDFIVKLTECVKAAKPKSQCGYKVDNV
DFHTVAHKISVGESNIDESRALVATILDEVKQWRDGITYHSSTPRNKSLMSLMVGWIPR
KAEKTKEILDNRIQRLELLLNQLNGVRGIDDYESLVRFFSENPHSAEYLESQCASDYIE
EKVMSVKRNYDKSLILGMVGLAVATGTFAYWYMRRSAAVELVEKQAKHKYNRDKRTGRL
MFDMDDRETYENFGPEYTDDVISAKMTKAQKERDSKKKGWKAGKINRPMRVFHQLYGVN
PLEFDEVVMRVGKLETEPWDVKELNVDAMMIELDDDYHILRDDRMFGKKVSLAFKKEGA
DEETIVNLTPHRSKMTSSMSLAPMGFPEEEGRWRQTGAPLIRKIEKEDEVEVQVAKPES
TNPYDHILVRLGRAHLGTRVLNCFFHGSKCVIPYHLAEKGDREESLVIATTRGQFDFGP
MKNIKCRKVTDYDITICPLPNDVQPFRSKIVMREPKLGEEVVIVCFTRINGKIVMKVSD
KSTTYPAGGQFAHLWAYKYDGQPGDCGGPIVATVDQKVVGFHSGVIRNSREEKLRAVYT
PVNQELLNCISGDIQMTDFWTFNPDLVEWNSVARVSTFFPMTKAINTITVQANEGEELI
DGNLMIVGYVNREVYHNHVIKGKRESFMRYCEQFPNCAFTKELRDQYLPSILSKPAFRK
GLLKYNEPVRVGSVNFSCLIRAYLKVEEMFEDLGFLEEAGPQWDPIEILDDLNKKAAMG
ALYQGKKQDWLKSIEPADFITAVRESFKHLAGGDVGIWSGSLKAELRPVEKVLEQKTRV
FTGAPIDLLLGGKILVDNFNHFFYFNHLKGPWTVGINKFNKGWDRLARYFNHSWNFIDC
DGSRFDTSLAPILFQLVCHMREKFGNFDDIERAALRNLYTQIVYTPILTIDGYITKKHR
GNNSGQPSTVVDNTIILMIVVEYCKAVMESEGRVMQFKYMCNGDDLILNVPDDEVSIVQ
SRFRELFSECGLDYNFDDVHKSIETIEYMSHSFMLKDGVYIPKLKKERIVAILEWERGD
EIMRTRSALNAAYIESYGYDDLMVEIERYAVFWATDKGCEYPLLDRKRVEGLYKDDYTD
INEEWLIGILPPSFEHCYVDTQTKDLRGREKLELRIESHDRTLQMQMKFPVTFVTGNLG
KLAEVKSILGIANDVMAKNIDLPEVQGTPEEIVRKKAQLAVKMTNSPVLVEDTCLCFNA
FNGLPGPYIKWFLKELGLDGVVKMLSAFGDKSAYALCTFAYVHNESSDPIVFKGVVNGE
IVPPRGNNGFGWDPIFKPDGCGCTFAEMPSSIKNDFSHRRRALEKVKLFLDNLMVKQEK
NKASVALTIDVQALNQEEIEAEITALKKLWKDNGPTRTRSSFEARRLRAPQVERVNELL
QKLKDEGLQTKKRPCGEPDDGEVVDDDSNDGDNQRSEKEVVDESQNNQQVDPRKLKFKI
RGDGNAIRRDDIDKIPTNALEIKKTFKPPKVSQSAYIWIPRSQRDNLTPDVIQNFLAYI
PPSHAIDNQLASGIEVENWAIEVAKAYGVNIQEFYRTVLPAWIVNCIVNGTSDERKNEK
SWRAVELNSQGEDVDDFEYPMEPMYKFALPTMRKVMRNFSSQAILMYQNSVAAGKAFVI
KAARNAGYTSIENKWLGIDFLAEAQLSQSQLDIKHQILAANVGRSKTRLFALAAPGDDG
NVDKERHTTHDVSANRHSYSGAAIE"
mat_peptide 1317..2402
/product="P1"
mat_peptide 2403..3284
/product="P3"
mat_peptide 3285..3440
/product="6K1"
mat_peptide 3441..5324
/product="CI"
mat_peptide 5325..5480
/product="6K2"
mat_peptide 5481..6035
/product="VPg"
mat_peptide 6036..6737
/product="NIa-Pro"
mat_peptide 6738..8243
/product="NIb"
mat_peptide 8244..8921
/product="Ham 1-like protein"
mat_peptide 8922..10022
/product="coat protein"
terminator 10288..10540
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 10578..10601
/label=RB
/note="RB"
regulatory complement(10666..10706)
/regulatory_class="terminator"
misc_feature 10716..10739
/label=assembly linker
/note="assembly linker"
CDS complement(10753..11565)
/label=KanR
/note="aminoglycoside phosphotransferase"
misc_feature 11667..11690
/label=assembly linker
/note="assembly linker"
CDS complement(11694..12353)
/label=pBBR1 Rep
/note="replication protein for the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica"
rep_origin complement(12354..13125)
/direction=LEFT
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"