Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V005495 | pGEM-7Zf(+) | 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
The pGEM-7Zf(+) and pGEM-7Zf(–) Vectors are derivatives of the pGEM-3Zf(+) Vector and contain the origin of replication of the filamentous phage f1. These plasmids serve as standard cloning vectors, as templates for in vitro transcription and can be used for the production of circular ssDNA. These plasmids contain SP6 and T7 RNA polymerase promoters flanking a region of multiple cloning sites within the α-peptide coding region of β-galactosidase. Insertional inactivation of the α-peptide allows recombinant clones to be identified directly by color screening on indicator plates when using appropriate E. coli strains (e.g., JM109). The multiple cloning region is unique and includes restriction sites for ApaI, AatII, SphI, XbaI, XhoI, EcoRI, KpnI, SmaI, ClaI, HindIII, BamHI, SacI, BstXI and NsiI. This arrangement is designed specifically for generating unidirectional deletions with the Erase-a-Base System. pGEM-7Zf(+) and pGEM-7Zf(–) Vectors are identical except for the orientation of the f1 origin.
- Vector Name:
- pGEM-7Zf(+)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 2997 bp
- Type:
- E. coli Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Promega
- Copy Number:
- High copy number
- Cloning Method:
- Enzyme digestion and ligation
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
pGEM-7Zf(+) 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.
pGEM-7Zf(+) vector Sequence
LOCUS 40924_21320 2997 bp DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2997)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 2997)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..2997
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(122..140)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(158..174)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(182..198)
/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)."
promoter complement(206..236)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(251..272)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(560..1148)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1322..2179)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2180..2284)
/label=AmpR promoter
rep_origin complement(2362..2817)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 2958..2974
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2981..2997
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"