The pSilencer vectors employ RNA polymerase III (pol III) promoters which generate large amounts of small RNA using relatively simple promoter and terminator sequences. They also include an antibiotic resistance gene that provides a mechanism to select for transfected cells that express the introduced DNA.
| 载体基本信息 | |||
| 载体名称 | pSilencer 3.1-H1 neo | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 4285 bp | 载体类型 | RNAi |
| 载体来源 | Thermo Fisher Scientific | 筛选标记 | Neomycin/G418(Geneticin) |
| 拷贝数 | High copy number | 启动子 | H1 |
| 克隆方法 | Enzyme Cut | 表达方法 | Transient |
pSilencer 3.1-H1 neo 载体载体图谱
pSilencer 3.1-H1 neo 载体载体序列
LOCUS Exported 4285 bp ds-DNA circular SYN 08-APR-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pSilencer 3.1-H1 neo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4285)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4285
/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"
polyA_signal 2625..2746
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(2922..3716)
/codon_start=1
/gene="aph(3)-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
promoter complement(3783..4099)
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 3797..3932
/note="SV40 ori"
/note="SV40 origin of replication"
