Basic Vector Information
pSilencer 5.1 Retro vectors harness retroviruses’ ability to stably integrate into the host genome of nearly all mitotically active cells and include a puromycin resistance gene for selection of cells that express the recombinant DNA. The pSilencer 5.1 Retro vectors enable researchers to study the long-term effects of reducing the expression of specific genes in cell culture models. This technology expands the capabilities of RNA interference research to address long-term alteration of gene expression.
- Vector Name:
- pSilencer 5.1-U6 Retro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6486 bp
- Type:
- RNAi
- Replication origin:
- ori
- Source/Author:
- Thermo Fisher Scientific
- Selection Marker:
- Puromycin
- Copy Number:
- High copy number
- Promoter:
- MSCV
- Cloning Method:
- Enzyme Cut
- Expression Method:
- Constiutive, Stable
pSilencer 5.1-U6 Retro vector Map
pSilencer 5.1-U6 Retro vector Sequence
LOCUS 40924_40382 6486 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 6486)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 6486)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6486
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 733..1249
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
misc_feature 1313..1654
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 1721..2137
/codon_start=1
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
/translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
primer_bind 2156..2172
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2246..2486)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
promoter 2580..3079
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 3104..3700
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
protein_bind complement(4289..4305)
/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(4313..4343)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4358..4379)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4667..5255)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5429..6286)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6287..6391)
/label=AmpR promoter
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