Basic Vector Information
The pGEM -7Zf(+) Vector (a) is a derivative of the pGEM -3Zf(+) Vector (a) and con- tains the origin of replication of the filamentous phage f1. The plasmid serves as a standard cloning vector, as a template for in vitro transcription, and can be used for the production of circular ssDNA. The plasmid contains SP6 and T7 RNA poly- merase promoters flanking a region of multiple cloning sites within the α -peptide coding region of β -galactosidase (1). Insertional inactivation of the α -peptide allows recombinant clones to be directly identified by color screening on indicator plates. The multiple cloning region is unique and includes restriction sites for Apa I, Aat II, Sph I, Xba I, Xho I, Eco R I, Kpn I, Sma I, Csp 45 I, Cla I, Hin d III, Bam H I, Sac I, Bst X I and Nsi I. This arrangement is designed specifically for generation of unidirec- tional deletions with Promega’s Erase-a-Base System. The polylinker contains restriction enzyme sites that produce 5 ́ overhangs or blunt ends (sensitive to Exonuclease III) flanked on both sides by blocks of restriction sites that generate 3 ́ overhangs (resistant to Exonuclease III).
- Vector Name:
- pEGM-7ZF(+)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 2997 bp
- Type:
- E. coli Cloning Vectors
- Replication origin:
- ori
- Cloning Method:
- Enzyme digestion and ligation
pEGM-7ZF(+) vector Vector Map
Plasmid Resuspension 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 ℃.
pEGM-7ZF(+) vector Sequence
LOCUS 40924_17279 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"
This page is informational only.