质粒编号
质粒名称
规格
价格
V005901
pgR106
5 μg (冻干粉)
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:
- pgR106
- 载体抗性:
- Kanamycin
- 载体长度:
- 11148 bp
- 载体类型:
- Transformation vector
- 复制子:
- ori
- 宿主:
- Plants
- 载体来源:
- Lu R, Malcuit I, Moffett P, Ruiz MT, Peart J, Wu AJ, Rathjen JP, Bendahmane A, Day L, Baulcombe DC.
- 启动子:
- CaMV 35S
pgR106 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。
pgR106 载体序列
LOCUS Exported 11148 bp DNA circular SYN 02-SEP-2024
DEFINITION Exported.
ACCESSION V005901
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11148)
AUTHORS Lu R, Malcuit I, Moffett P, Ruiz MT, Peart J, Wu AJ, Rathjen JP,
Bendahmane A, Day L, Baulcombe DC.
TITLE High throughput virus-induced gene silencing implicates heat shock
protein 90 in plant disease resistance
JOURNAL EMBO J. 22 (21), 5690-5699 (2003)
PUBMED 14592968
REFERENCE 2 (bases 1 to 11148)
AUTHORS Lu R, Malcuit I, Moffett P, Ruiz T, Peart J, Wu A-J., Rathjen J,
Bendahmane A, Day L, Baulcombe D.
TITLE Direct Submission
JOURNAL Submitted (13-MAY-2003) Department of Plant Pathology, University of
California Riverside, 900 University Avenue, Riverside, CA 92521,
USA
REFERENCE 3 (bases 1 to 11148)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11148)
TITLE Direct Submission
REFERENCE 5 (bases 1 to 11148)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "EMBO J.";
date: "2003"; volume: "22"; issue: "21"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(13-MAY-2003) Department of Plant Pathology, University of
California Riverside, 900 University Avenue, Riverside, CA 92521,
USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT SGRef: number: 4; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..11148
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 10142..10917
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 558..582
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
protein_bind 821..842
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 857..887
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 895..911
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 919..935
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 956..974
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 1082..1427
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 1510..5877
/codon_start=1
/label=RNA replication protein
/note="RNA replication protein from Potato virus X (strain
X3). Accession#: P17779"
/translation="MAKVREVYQSFTDSTTKTLIQDEAYRNIRPIMEKHKLANPYAQTV
EAANDLEGFGIATNPYSIELHTHAAAKTIENKLLEVLGSILPQEPVTFMFLKPRKLNYM
RRNPRIKDIFQNVAIEPRDVARYPKETIIDKLTEITTETAYISDTLHFLDPSYIVETFQ
NCPKLQTLYATLVLPVEAAFKMESTHPNIYSLKYFGDGFQYIPGNHGGGAYHHEFAHLQ
WLKVGKIKWRDPKDSFLGHLNYTTEQVEMHTVTVQLQESFAANHLYCIRRGDLLTPEVR
TFGQPDRYVIPPQIFLPKVHNCKKPILKKTMMQLFLYVRTVKVAKNCDIFAKVRQLIKS
SDLDKYSAVELVYLVSYMEFLADLQATTCFSDTLSGGLLTKTLAPVRAWIQEKKMQLFG
LEDYAKLVKAVDFHPVDFSFKVETWDFRFHPLQAWKAFRPREVSDVEEMESLFSDGDLL
DCFTRMPAYAVNAEEDLAAIRKTPEMDVGQEVKEPAGDRNQYSNPAETFLNKLHRKHSR
EVKHQAAKKAKRLAEIQESMRAEGDAEPNEISGGMGAIPSNAELPGTNDARQELTLPTT
KPVPARWEDASFTDSSVEEEQVKPLGKETVETATQQVIEGLPWKHWIPQLNAVGFKALE
IQRDRSGTMIMPITEMVSGLEKEDFPEGTPKELARELFAMNRSPATIPLDLLRARDYGS
DVKNKRIGAITKTQATSWGEYLTGKIESLTERKVATCVIHGAGGSGKSHAIQKALREIG
KGSDITVVLPTNELRLDWSKKVPNTEPYMFKTYEKALIGGTGSIVIFDDYSKLPPGYIE
ALVCFYSKIKLIILTGDSRQSVYHETAEDASIRHLGPATEYFSKYCRYYLNATHRNKKD
LANMLGVYSERTGVTEISMSAEFLEGIPTLVPSDEKRKLYMGTGRNDTFTYAGCQGLTK
PKVQIVLDHNTQVCSANVMYTALSRATDRIHFVNTSANSSAFWEKLDSTPYLKTFLSVV
REQALREYEPAEAEPIQEPEPQTHMCVENEESVLEEYKEELLEKFDREIHSESHGHSNC
VQTEDTTIQLFSHQQAKDETLLWATIDARLKTSNQETNFREFLSKKDIGDVLFLNYQKA
MGLPKERIPFSQEVWEACAHEVQSKYLSKSKCNLINGTVRQSPDFDENKIMVFLKSQWV
TKVEKLGLPKIKPGQTIAAFYQQTVMLFGTMARYMRWFRQAFQPKEVFINCETTPDDMS
AWALNNWNFSRPSLANDYTAFDQSQDGAMLQFEVLKAKHHCIPEEIIQAYIDIKTNAQI
FLGTLSIMRLTGEGPTFDANTECNIAYTHTKFDIPAGTAQVYAGDDSALDCVPEVKHSF
HRLEDKLLLKSKPVITQQKKGSWPEFCGWLITPKGVMKDPIKLHVSLKLAEAKGELKKC
QDSYEIDLSYAYDHKDSLHDLFDEKQCQAHTLTCRTLIKSGRGTVSLSRLRNFL"
CDS 5911..6585
/codon_start=1
/label=Movement and silencing protein TGBp1
/note="Movement and silencing protein TGBp1 from Potato
virus X (strain X3). Accession#: P17780"
/translation="MDILISSLKSLGYSRTSKSLDSGPLVVHAVAGAGKSTALRKLILR
HPTFTVHTLGVPDKVSIRTRGIQKPGPIPEGNFAILDEYTLDNTTRNSYQALFADPYQA
PEFSLEPHFYLETSFRVPRKVADLIAGCGFDFETNSPEEGHLEITGIFKGPLLGKVIAI
DEESETTLSRHGVEFVKPCQVTGLEFKVVTIVSAAPIEEIGQSTAFYNAITRSKGLTYV
RAG"
misc_feature 7093..7125
/label=multiple cloning site
/note="multiple cloning site"
misc_feature 7126..8002
/label=Potato virus X strain UK3 3'-sequence
/note="Potato virus X strain UK3 3'-sequence"
terminator 8027..8279
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
misc_feature 8285..8452
/label=shuttle vector pSK5640 sequence
/note="shuttle vector pSK5640 sequence"
misc_feature 8460..8482
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA
(truncated)"
rep_origin complement(8549..9137)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9238..10053)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
mobile_element 10147..10914
/mobile_element_type="insertion sequence:IS1"
/label=IS1
/note="prokaryotic transposable element"
rep_origin join(11120..11148,1..407)
/label=pSa ori
/note="origin of replication from bacterial plasmid pSa"