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。