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.
| 载体基本信息 | |||
| 载体名称 | pGEM-7Zf(+) | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 2997 bp | 载体类型 | E. coli Expression Vectors |
| 载体来源 | Promega | 拷贝数 | High copy number |
| 克隆方法 | Enzyme digestion and ligation | 5引物 | M13 fwd |
| 3引物 | M13 rev | ||
pGEM-7Zf(+) 载体载体图谱
pGEM-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"
