Basic Vector Information
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.
- Vector Name:
- pSilencer 3.0-H1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 2795 bp
- Type:
- RNAi
- Replication origin:
- ori
- Source/Author:
- Thermo Fisher Scientific
- Copy Number:
- High copy number
- Promoter:
- H1
- Cloning Method:
- Enzyme Cut
- Expression Method:
- Transient
pSilencer 3.0-H1 vector Map
pSilencer 3.0-H1 vector Sequence
LOCUS 40924_40357 2795 bp DNA circular SYN 08-APR-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2795)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 2795)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..2795
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 402..492
/label=H1 promoter
/note="human H1 RNA promoter"
primer_bind complement(574..590)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(598..614)
/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(622..652)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(667..688)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(976..1564)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1738..2595)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2596..2700)
/label=AmpR promoter
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