| 载体基本信息 | |||
| 载体名称 | NLSx3-CRY2PHR-p65 | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 8026 bp | 载体类型 | Mammalian Expression |
| 筛选标记 | Neomycin (select with G418) | 拷贝数 | High Copy |
| 启动子 | CMV | 克隆方法 | Restriction Enzyme |
| 5'引物 | CMV-F | 3'引物 | BGH Reverse |
NLSx3-CRY2PHR-p65 载体载体图谱
NLSx3-CRY2PHR-p65 载体载体序列
LOCUS Exported 8026 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Photoactivatable transcription system. Expression of activator
probe, which includes the photolyase homology region of CRY2
(CRY2PHR) and the transcriptional activator domain..
ACCESSION .
VERSION .
KEYWORDS NLSx3-CRY2PHR-p65
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8026)
AUTHORS Nihongaki Y, Yamamoto S, Kawano F, Suzuki H, Sato M
TITLE CRISPR-Cas9-based Photoactivatable Transcription System.
JOURNAL Chem Biol. 2015 Feb 19;22(2):169-74. doi:
10.1016/j.chembiol.2014.12.011. Epub 2015 Jan 22.
PUBMED 25619936
REFERENCE 2 (bases 1 to 8026)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1016/j.chembiol.2014.12.011"; journalName: "Chem Biol"; date:
"2015-02-19- 19"; volume: "22"; issue: "2"; pages: "169-74"
FEATURES Location/Qualifiers
source 1..8026
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 908..917
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 923..943
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 959..979
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1085..2578
/codon_start=1
/product="N-terminal photolyase homology region of
Arabidopsis thaliana cryptochrome 2"
/label=CRY2PHR
/note="binds to CIB1 upon exposure to blue light (Kennedy
et al., 2010)"
/translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
CDS 2636..2662
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 3092..3463
/codon_start=1
/product="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
/label=RelA (p65) AD
/translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
EDFSSIADMDFSALLSQISS"
CDS 3524..3565
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 3575..3592
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(3615..3632)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3621..3845
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3891..4319
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3978..3997)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4188..4209
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4328..4348)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4333..4662
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4513..4648
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4575..4594
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4729..5523
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4783..4802)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5393..5412
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5699..5820
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5736..5755)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5790..5809
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5869..5885)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5869..5885)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5882..5904)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5893..5909
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(5917..5947)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5962..5983
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6100..6117)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6271..6859)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6351..6370)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7030..7890)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7653..7672
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7891..7995)
/gene="bla"
/label=AmpR promoter
我们不提供该载体，载体信息仅供查询和参考。