The pSilencer 4.1-CMV siRNA Expression vectors are linearized with both BamH 1 and Hind III to facilitate directional cloning. They are also 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 template insert after ligation, reducing the time and effort required to screen clones.
| 载体基本信息 | |||
| 载体名称 | pSilencer 4.1-CMV neo | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 4944 bp | 载体类型 | RNAi |
| 载体来源 | Thermo Fisher Scientific | 拷贝数 | High copy number |
| 启动子 | CMV | 克隆方法 | Enzyme Cut |
| 表达方法 | Transient | ||
pSilencer 4.1-CMV neo 载体载体图谱
pSilencer 4.1-CMV neo 载体载体序列
LOCUS Exported 4944 bp ds-DNA circular SYN 08-APR-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pSilencer 4.1-CMV neo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4944)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4944
/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 complement(536..739)
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
enhancer 740..1043
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter complement(1165..1183)
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(1195..1213)
/note="SP6 promoter"
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1233..1249)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1257..1273
/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(1281..1311)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 1326..1347
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1635..2223)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2394..3254)
/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 3284..3405
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(3581..4375)
/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(4442..4758)
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 4456..4591
/note="SV40 ori"
/note="SV40 origin of replication"
