Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007993 | pDhtSK | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pDhtSK
- Antibiotic Resistance:
- Kanamycin
- Length:
- 7474 bp
- Type:
- UNVERIFIED: Cloning vector
- Replication origin:
- ori
- Source/Author:
- Feng J.
pDhtSK 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.
pDhtSK vector Sequence
LOCUS 40924_14705 7474 bp DNA circular SYN 18-DEC-2018
DEFINITION UNVERIFIED: Cloning vector pDhtSK, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7474)
AUTHORS Feng J.
TITLE Direct Submission
JOURNAL Submitted (29-JAN-2018) Sun Yat-sen Memorial Hospital, Sun Yat-sen
University, Department of Dermatology, 107 West Yanjiang Rd,
Guangzhou, Guangdong 510120, China
REFERENCE 2 (bases 1 to 7474)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7474)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
(29-JAN-2018) Sun Yat-sen Memorial Hospital, Sun Yat-sen University,
Department of Dermatology, 107 West Yanjiang Rd, Guangzhou,
Guangdong 510120, China"
COMMENT SGRef: number: 2; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
GenBank staff is unable to verify sequence and/or annotation
provided by the submitter.
FEATURES Location/Qualifiers
source 1..7474
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal complement(77..251)
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
protein_bind 552..573
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 588..618
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 626..642
/label=lac operator
/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 650..666
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 687..705
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
misc_feature 718..825
/label=MCS
/note="pBluescript multiple cloning site"
promoter complement(834..852)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(862..878)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 1171..1195
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 2495..3121
/codon_start=1
/label=pVS1 StaA
/note="stability protein from the Pseudomonas plasmid pVS1
(Heeb et al., 2000)"
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 3558..4622
/codon_start=1
/label=pVS1 RepA
/note="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
rep_origin 4691..4885
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
misc_feature 5229..5369
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(5555..6143)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6233..7024)
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 7449..7473
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"