Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V005290 | pJRD215 | 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:
- pJRD215
- Antibiotic Resistance:
- Kanamycin
- Length:
- 10316 bp
- Type:
- Expression vector
- Replication origin:
- RSF1010 oriV
- Source/Author:
- Davison J, Heusterspreute M, Chevalier N, Ha-Thi V, Brunel F.
- Promoter:
- tet
pJRD215 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.
pJRD215 vector Sequence
LOCUS 40924_26336 10316 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pJRD215, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10316)
AUTHORS Davison J, Heusterspreute M, Chevalier N, Ha-Thi V, Brunel F.
TITLE Vectors with restriction site banks. V. pJRD215, a wide-host-range
cosmid vector with multiple cloning sites
JOURNAL Gene 51 (2-3), 275-280 (1987)
PUBMED 3036654
REFERENCE 2 (bases 1 to 10316)
AUTHORS Piatkov K.
TITLE Direct Submission
JOURNAL Submitted (14-JUN-2012) Biology, California Institute of Technology,
1200 E. California Blvd., 147-75, Pasadena, CA 91125, USA
REFERENCE 3 (bases 1 to 10316)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10316)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene 51
(2-3), 275-280 (1987)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(14-JUN-2012) Biology, California Institute of Technology, 1200 E.
California Blvd., 147-75, Pasadena, CA 91125, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10316
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 13..41
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
CDS 253..1056
/codon_start=1
/product="Sm resistance protein A"
/label=Sm resistance protein A
/protein_id="AFQ55241.1"
/translation="MNRTNIFFGESHSDWLPVRGGESGDFVFRRGDGHAFAKIAPASRR
GELAGERDRLIWLKGRGVACPEVINWQEEQEGACLVITAIPGVPAADLSGADLLKAWPS
MGQQLGAVHSLSVDQCPFERRLSRMFGRAVDVVSRNAVNPDFLPDEDKSTPLHDLLARV
ERELPVRLDQERTDMVVCHGDPCMPNFMVDPKTLQCTGLIDLGRLGTADRYADLALMIA
NAEENWAAPDEAERAFAVLFNVLGIEAPDRERLAFYLRLDPLTWG"
CDS 1056..1892
/codon_start=1
/product="Sm resistance protein B"
/label=Sm resistance protein B
/protein_id="AFQ55242.1"
/translation="MFMPPVFPAHWHVSQPVLIADTFSSLVWKVSLPDGTPAIVKGLKP
IEDIADELRGADYLVWRNGRGAVRLLGRENNLMLLEYAGERMLSHIVAEHGDYQATEIA
AELMAKLYAASEEPLPSALLPIRDRFAALFQRARDDQNAGCQTDYVHAAIIADQMMSNA
SELRGLHGDLHHENIMFSSRGWLVIDPVGLVGEVGFGAANMFYDPADRDDLCLDPRRIA
QMADAFSRALDVDPRRLLDQAYAYGCLSAAWNADGEEEQRDLAIAAAIKQVRQTSY"
rep_origin complement(2536..2930)
/direction=LEFT
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
CDS complement(2957..3241)
/codon_start=1
/product="mobilization protein C"
/label=mobilization protein C
/protein_id="AFQ55243.1"
/translation="MVKGSNKAADRLAKLEEQRARINAEIQRVRAREQQQERKNETRRK
VLVGAMILAKVNSSEWPEDRLMAAMDAYLERDHDRALFGLPPRQKDEPG"
oriT 3272..3359
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS 4188..4601
/codon_start=1
/product="mobilization protein B"
/label=mobilization protein B
/protein_id="AFQ55245.1"
/translation="MNAIDRVKKSRGINELAEQIEPLAQSMATLADEARQVMSQTQQAS
EAQAAEWLKAQRQTGAAWVELAKELREVAAEVSSAAQSARSASRGWHWKLWLTVMLASM
MPTVVLLIASLLLLDLTPLTTEDGSIWLRLVAR"
CDS 4598..5566
/codon_start=1
/label=RSF1010 RepB
/note="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
CDS 5630..5842
/codon_start=1
/product="unknown"
/label=unknown
/note="E"
/protein_id="AFQ55247.1"
/translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
LNLDGCTLSLFREDKPFGPGKFLGD"
CDS 5844..6050
/codon_start=1
/product="repressor protein F"
/label=repressor protein F
/protein_id="AFQ55248.1"
/translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
EALRECLEELRAAQGGGSDPASA"
CDS 6080..6916
/codon_start=1
/label=RSF1010 RepA
/note="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
CDS 6906..7754
/codon_start=1
/label=RSF1010 RepC
/note="replication protein C of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/translation="VVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
AG"
CDS complement(8282..9073)
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
misc_feature 9457..9856
/label=cos
/note="lambda cos site; allows packaging into phage lambda
particles"