The pSilencer siRNA Expression Vectors are linearized with both BamH 1 and Hind III to facilitate directional cloning. They are purified to remove the digested insert so that it cannot re-ligate with the vector. This greatly increases the percentage of clones bearing the hairpin siRNA-coding insert after ligation, reducing the time and effort required to screen clones. Both pSilencer 2.0-U6 and pSilencer 3.0-H1 are linearized with the same restriction enzymes, so that a given hairpin siRNA insert can be subcloned into either vector using the 5 overhangs left by restriction enzyme digestion.
| 载体基本信息 | |||
| 载体名称 | pSilencer 3.0-H1 | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 2795 bp | 载体类型 | RNAi |
| 载体来源 | Thermo Fisher Scientific | 拷贝数 | High copy number |
| 启动子 | H1 | 克隆方法 | Enzyme Cut |
| 表达方法 | Transient | ||
pSilencer 3.0-H1 载体载体图谱
pSilencer 3.0-H1 载体载体序列
LOCUS Exported 2795 bp ds-DNA circular SYN 08-APR-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pSilencer 3.0-H1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2795)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..2795
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 379..395
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 402..492
/note="H1 promoter"
/note="human H1 RNA promoter"
primer_bind complement(574..590)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 598..614
/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(622..652)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 667..688
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(976..1564)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1735..2595)
/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(2596..2700)
/gene="bla"
/note="AmpR promoter"
