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).
| 载体基本信息 | |||
| 载体名称 | pEGM-7ZF(+) | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 2997 bp | 载体类型 | E. coli Cloning Vectors |
| 克隆方法 | Enzyme digestion and ligation | ||
pEGM-7ZF(+) 载体载体图谱
pEGM-7ZF(+) 载体载体序列
LOCUS Exported 2997 bp ds-DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2997)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..2997
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(122..140)
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(158..174)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 182..198
/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)."
promoter complement(206..236)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 251..272
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(560..1148)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1319..2179)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2180..2284)
/gene="bla"
/note="AmpR promoter"
rep_origin complement(2362..2817)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(2794..162)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITPSYLGDTIEYSSYASNALGALRIQAYRFRTRGTEFLESRG
ACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPS
QQLRSLNGEWTRPVAAH"
primer_bind 2958..2974
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 2981..2
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
