| 质粒名称: | pcDNA3-TLR4-YFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 13018 |
| 存储实验室: | Doug Golenbock |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3-YFP |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6100 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Toll-Like Receptor 4,TLR4,TOLL,CD284 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2517 bp |
| 基因突变: | -- |
| GenBank编号 | BC117422 |
| 启动子 | -- |
| 标签/融合蛋白: | YFP（主干上的C终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI(未破坏) |
| 3' 克隆位点: | XhoI(未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | GTCTTGTAGTTGCCGTCGTC |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3-TLR4-YFP was a gift from Doug Golenbock (Addgene plasmid # 13018 ; http://n2t.net/addgene:13018 ; RRID:Addgene_13018) |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pcDNA3-TLR4-YFP 8618 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3-TLR4-YFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8618)
TITLE TLR Signaling Plasmids
REFERENCE 2 (bases 1 to 8618)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8618
/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"
CDS 3436..4155
/codon_start=1
/product="enhanced YFP"
/label=EYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3481..3502)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3742..3761)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4089..4110
/label=EGFP-C
/note="EGFP, forward primer"
promoter complement(4171..4189)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(4172..4189)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(4209..4226)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4215..4439
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4485..4913
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4572..4591)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4782..4803
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4922..4942)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4927..5256
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5107..5242
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5169..5188
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5323..6117
/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(5377..5396)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5987..6006
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6291..6412
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6328..6347)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6382..6401
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6461..6477)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6461..6477)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6474..6496)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6485..6501
/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(6509..6539)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6554..6575
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6692..6709)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6863..7451)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6943..6962)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7622..8482)
/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 8245..8264
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8483..8587)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
901 gagctcggat ccatgatgtc tgcctcgcgc ctggctggga ctctgatccc agccatggcc
961 ttcctctcct gcgtgagacc agaaagctgg gagccctgcg tggaggtggt tcctaatatt
1021 acttatcaat gcatggagct gaatttctac aaaatccccg acaacctccc cttctcaacc
1081 aagaacctgg acctgagctt taatcccctg aggcatttag gcagctatag cttcttcagt
1141 ttcccagaac tgcaggtgct ggatttatcc aggtgtgaaa tccagacaat tgaagatggg
1201 gcatatcaga gcctaagcca cctctctacc ttaatattga caggaaaccc catccagagt
1261 ttagccctgg gagccttttc tggactatca agtttacaga agctggtggc tgtggagaca
1321 aatctagcat ctctagagaa cttccccatt ggacatctca aaactttgaa agaacttaat
1381 gtggctcaca atcttatcca atctttcaaa ttacctgagt atttttctaa tctgaccaat
1441 ctagagcact tggacctttc cagcaacaag attcaaagta tttattgcac agacttgcgg
1501 gttctacatc aaatgcccct actcaatctc tctttagacc tgtccctgaa ccctatgaac
1561 tttatccaac caggtgcatt taaagaaatt aggcttcata agctgacttt aagaaataat
1621 tttgatagtt taaatgtaat gaaaacttgt attcaaggtc tggctggttt agaagtccat
1681 cgtttggttc tgggagaatt tagaaatgaa ggaaacttgg aaaagtttga caaatctgct
1741 ctagagggcc tgtgcaattt gaccattgaa gaattccgat tagcatactt agactactac
1801 ctcgatgata ttattgactt atttaattgt ttgacaaatg tttcttcatt ttccctggtg
1861 agtgtgacta ttgaaagggt aaaagacttt tcttataatt tcggatggca acatttagaa
1921 ttagttaact gtaaatttgg acagtttccc acattgaaac tcaaatctct caaaaggctt
1981 actttcactt ccaacaaagg tgggaatgct ttttcagaag ttgatctacc aagccttgag
2041 tttctagatc tcagtagaaa tggcttgagt ttcaaaggtt gctgttctca aagtgatttt
2101 gggacaacca gcctaaagta tttagatctg agcttcaatg gtgttattac catgagttca
2161 aacttcttgg gcttagaaca actagaacat ctggatttcc agcattccaa tttgaaacaa
2221 atgagtgagt tttcagtatt cctatcactc agaaacctca tttaccttga catttctcat
2281 actcacacca gagttgcttt caatggcatc ttcaatggct tgtccagtct cgaagtcttg
2341 aaaatggctg gcaattcttt ccaggaaaac ttccttccag atatcttcac agagctgaga
2401 aacttgacct tcctggacct ctctcagtgt caactggagc agttgtctcc aacagcattt
2461 aactcactct ccagtcttca ggtactaaat atgagccaca acaacttctt ttcattggat
2521 acgtttcctt ataagtgtct gaactccctc caggttcttg attacagtct caatcacata
2581 atgacttcca aaaaacagga actacagcat tttccaagta gtctagcttt cttaaatctt
2641 actcagaatg actttgcttg tacttgtgaa caccagagtt tcctgcaatg gatcaaggac
2701 cagaggcagc tcttggtgga agttgaacga atggaatgtg caacaccttc agataagcag
2761 ggcatgcctg tgctgagttt gaatatcacc tgtcagatga ataagaccat cattggtgtg
2821 tcggtcctca gtgtgcttgt agtatctgtt gtagcagttc tggtctataa gttctatttt
2881 cacctgatgc ttcttgctgg ctgcataaag tatggtagag gtgaaaacat ctatgatgcc
2941 tttgttatct actcaagcca ggatgaggac tgggtaagga atgagctagt aaagaattta
3001 gaagaagggg tgcctccatt tcagctctgc cttcactaca gagactttat tcccggtgtg
3061 gccattgctg ccaacatcat ccatgaaggt ttccataaaa gccgaaaggt gattgttgtg
3121 gtgtcccagc acttcatcca gagccgctgg tgtatctttg aatatgagat tgctcagacc
3181 tggcagtttc tgagcagtcg tgctggtatc atcttcattg tcctgcagaa ggtggagaag
3241 accctgctca ggcagcaggt ggagctgtac cgccttctca gcaggaacac ttacctggag
3301 tgggaggaca gtgtcctggg gcggcacatc ttctggagac gactcagaaa agccctgctg
3361 gatggtaaat catggaatcc agaaggaaca gtgggtacag gatgcaattg gcaggaagca
3421 acatctatcc tcgagatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
3481 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
3541 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
3601 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgaagtgctt cgcccgctac
3661 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
3721 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
3781 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
3841 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
3901 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
3961 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
4021 ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag
4081 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
4141 gagctgtaca agtaatctag agggccctat tctatagtgt cacctaaatg ctagagctcg
4201 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
4261 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
4321 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
4381 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
4441 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
4501 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
4561 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
4621 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
4681 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
4741 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
4801 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
4861 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
4921 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
4981 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
5041 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
5101 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
5161 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
5221 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
5281 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
5341 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
5401 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
5461 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
5521 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
5581 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
5641 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
5701 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
5761 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
5821 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
5881 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
5941 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
6001 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
6061 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
6121 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
6181 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
6241 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
6301 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
6361 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
6421 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
6481 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
6541 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
6601 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
6661 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
6721 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
6781 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
6841 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
6901 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
6961 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
7021 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
7081 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
7141 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
7201 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
7261 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
7321 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
7381 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
7441 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
7501 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
7561 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
7621 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
7681 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
7741 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
7801 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
7861 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
7921 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
7981 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
8041 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
8101 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
8161 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
8221 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
8281 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
8341 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
8401 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
8461 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
8521 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
8581 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
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