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.
| 载体基本信息 | |||
| 载体名称 | pSilencer 5.1-U6 Retro | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 6486 bp | 载体类型 | RNAi |
| 载体来源 | Thermo Fisher Scientific | 筛选标记 | Puromycin |
| 拷贝数 | High copy number | 启动子 | U6 |
| 克隆方法 | Enzyme Cut | 表达方法 | Constiutive, Stable |
pSilencer 5.1-U6 Retro 载体载体图谱
pSilencer 5.1-U6 Retro 载体载体序列
LOCUS Exported 6486 bp ds-DNA circular SYN 08-APR-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pSilencer 5.1-U6 Retro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6486)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6486
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 733..1249
/note="5 LTR"
/note="5 long terminal repeat from murine embryonic stem
cell virus"
misc_feature 1313..1654
/note="MESV Psi"
/note="packaging signal of murine embryonic stem cell
virus"
CDS 1721..2137
/note="gag (truncated)"
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 2156..2172
/note="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(2246..2486)
/note="U6 promoter"
/note="RNA polymerase III promoter for human U6 snRNA"
promoter 2580..3079
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 3104..3703
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
protein_bind 4289..4305
/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(4313..4343)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 4358..4379
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4667..5255)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5426..6286)
/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(6287..6391)
/gene="bla"
/note="AmpR promoter"
