pSilencer 3.0-H1 vector (V012700)

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

Copy Sequence View 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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