Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V005901 | pgR106 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pgR106
- Antibiotic Resistance:
- Kanamycin
- Length:
- 11148 bp
- Type:
- Transformation vector
- Replication origin:
- ori
- Host:
- Plants
- Source/Author:
- Lu R, Malcuit I, Moffett P, Ruiz MT, Peart J, Wu AJ, Rathjen JP, Bendahmane A, Day L, Baulcombe DC.
- Promoter:
- CaMV 35S
pgR106 vector Map
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5. Store the plasmid at -20 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pgR106 vector Sequence
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"