Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V014703 | pHRW32 | 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:
- pHRW32
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10942 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Yeast
- Selection Marker:
- Zeo
- Promoter:
- TEF
- Growth Temperature:
- 37℃
pHRW32 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.
pHRW32 vector Sequence
LOCUS Exported 10942 bp DNA circular SYN 12-SEP-2024
DEFINITION S22409110001-PHRW32_for.ref.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10942)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..10942
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 1265..1276
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 1922..2419
/codon_start=1
/product="13 tandem Myc epitope tags"
/label=13xMyc
/translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
EDL"
terminator 2612..2859
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(2882..2898)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2906..2922
/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)."
promoter complement(2930..2960)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2975..2996
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3284..3872)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4043..4903)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4904..5008)
/gene="bla"
/label=AmpR promoter
promoter 5216..5559
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
terminator 5940..6137
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
rep_origin 6480..7317
/label=ARS1
/note="S. cerevisiae autonomously replicating sequence
ARS1/ARS416"
misc_feature 7453..7745
/label=CEN4
/note="S. cerevisiae CEN4 centromere"
primer_bind 8502..8518
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(8546..9553)
/codon_start=1
/product="tetracycline-controlled transactivator,
comprising a fusion of the tetracycline repressor TetR with
the C-terminal activation domain of herpes simplex virus
VP16"
/label=tTA
/note="In the Tet-Off(R) system, tTA binds to the
Tet-responsive element and stimulates transcription only in
the absence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
promoter complement(9659..9862)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
enhancer 9863..10242
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
protein_bind 10597..10867
/label=tetracycline response element
/bound_moiety="tetracycline repressor TetR"
/note="contains seven copies of the tetracycline operator
tetO"