The pGEM-11Zf(+) Vector can be used as a standard cloning vector and as a template for in vitro transcription and the production of ssDNA. The plasmid contains T7 and SP6 RNA polymerase promoters flanking a multiple cloning region within the alpha-peptide coding region of beta-galactosidase. Insertional inactivation of the alpha-peptide allows recombinant clones to be identified directly by color screening on indicator plates when using appropriate E.coli strains (e.g., JM109, DH5alpha, XL1-Blue). The multiple cloning region contains unique restriction sites for SfiI, SacI, EcoRI, SalI, XhoI, BamHI, ApaI, XbaI, NotI, SphI, NsiI and HindIII.
| 载体基本信息 | |||
| 载体名称 | pEGM-11ZF(+) | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 3221 bp | 载体类型 | E. coli Cloning Vectors |
| 克隆方法 | Enzyme digestion and ligation | ||
pEGM-11ZF(+) 载体载体图谱
pEGM-11ZF(+) 载体载体序列
LOCUS Exported 3221 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 3221)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3221
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(92..110)
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(128..144)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 152..168
/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(176..206)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 221..242
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(530..1118)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1289..2149)
/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(2150..2254)
/gene="bla"
/note="AmpR promoter"
rep_origin complement(2586..3041)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(3018..132)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/note="lacZ-alpha"
/translation="MTMITPSYLGDTIEYSSLCMRPHLEGPDPSRSTNSSSADLANSPY
SESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEW
TRPVAAH"
primer_bind 3182..3198
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 3205..2
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
