Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V010923 | pRSFDuet-1 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
The pRSFDuet-1 vector contains an RSF 1030 origin and dual promoters, allowing the simultaneous expression of two genes. This makes it highly efficient for co-expressing two proteins and is ideal for studying protein-protein interactions and complex biological pathways.
The pRSFDuet-1 vector is suitable for cases where one is studying protein complexes or interactions requiring co-expression of two genes.
- Vector Name:
- pRSFDuet-1
- Antibiotic Resistance:
- Kanamycin
- Length:
- 3829 bp
- Type:
- pET & Duet Vectors (Novagen)
- Replication origin:
- RSF ori
- Source/Author:
- Novagen (EMD Millipore)
- Copy Number:
- High copy number
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pRSFDuet-1 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.
References
- Chen Y, Ma B, Cao S, Wu X, Xu Y. Efficient synthesis of Ibrutinib chiral intermediate in high space-time yield by recombinant E. coli co-expressing alcohol dehydrogenase and glucose dehydrogenase. RSC Adv. 2019 Jan 18;9(4):2325-2331. doi: 10.1039/c8ra08100j. PMID: 35516114; PMCID: PMC9059822.
pRSFDuet-1 vector Sequence
LOCUS Exported 3829 bp DNA circular SYN 10-SEP-2025
DEFINITION Bacterial vector with an RSF 1030 origin for the co-expression of
two genes.
ACCESSION .
VERSION .
KEYWORDS pRSFDuet-1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3829)
AUTHORS Novagen (EMD Millipore)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 3829)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 3829)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3829
/mol_type="other DNA"
/organism="synthetic DNA construct"
source join(48..3829,1..47)
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 31..49
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 50..74
/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)."
RBS 89..111
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 118..120
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 130..147
/label=6xHis
/note="6xHis affinity tag"
promoter 261..279
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 280..304
/label=lac repressor encoded by lacI binding site
/bound_moiety="lac repressor encoded by lacI"
/note="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)."
RBS 333..338
/note="ribosome binding site"
CDS 347..349
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
CDS 413..457
/label=S-Tag
/note="affinity and epitope tag derived from pancreatic
ribonuclease A"
terminator 509..556
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(786..1601)
/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"
promoter complement(1602..1693)
/label=AmpR promoter
rep_origin complement(1709..2458)
/direction=LEFT
/label=RSF ori
/note="Plasmids containing the RSF 1030 origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
protein_bind complement(2620..2641)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(2654..3736)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(3737..3814)
/label=lacI promoter