| 质粒名称: | pmirGLO-3'UTR-TNF |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 153069 |
| 存储实验室: | Silvia Monticelli |
|
载体骨架基本信息 |
|
| 载体名称: | pmirGLO Dual-Luciferase |
| 载体出品公司: | Promega |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7345 bp |
| 完整质粒大小: | 8119 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，荧光素酶 |
| 筛选标记: | -- |
| 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: | 3' UTR region of TNF gene |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 774 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | PGK |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI(未破坏) |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | AGAAGCTGCGCGGTGGTGTTGTG |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pmirGLO-3'UTR-TNF was a gift from Silvia Monticelli (Addgene plasmid # 153069 ; http://n2t.net/addgene:153069 ; RRID:Addgene_153069) |
| 参考文献部分: | A molecular network regulating the proinflammatory phenotype of human memory T lymphocytes. Emming S, Bianchi N, Polletti S, Balestrieri C, Leoni C, Montagner S, Chirichella M, Delaleu N, Natoli G, Monticelli S. Nat Immunol. 2020 Apr;21(4):388-399. doi: 10.1038/s41590-020-0622-8. Epub 2020 Mar 16. 10.1038/s41590-020-0622-8 PubMed 32205878 |
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 pmirGLO-3'UTR-TNF 8119 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Human TNF 3'UTR region cloned downstream of luciferase reporter
gene.
ACCESSION .
VERSION .
KEYWORDS pmirGLO-3'UTR-TNF
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8119)
AUTHORS Emming S, Bianchi N, Polletti S, Balestrieri C, Leoni C, Montagner
S, Chirichella M, Delaleu N, Natoli G, Monticelli S
TITLE A molecular network regulating the proinflammatory phenotype of
human memory T lymphocytes.
JOURNAL Nat Immunol. 2020 Apr;21(4):388-399. doi: 10.1038/s41590-020-0622-8.
Epub 2020 Mar 16.
PUBMED 32205878
REFERENCE 2 (bases 1 to 8119)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8119
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 89..108
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
regulatory 197..206
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 203..1855
/codon_start=1
/product="firefly luciferase"
/label=luciferase
/note="synthetic luc2 version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(2737..2755)
/label=T7 Term
/note="T7 terminator, reverse primer"
polyA_signal 2792..2913
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2803..2822)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2857..2876
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 3158..3515
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3181..3201)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3366..3501
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3428..3447
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
regulatory 3560..3569
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3566..4498
/codon_start=1
/product="Renilla luciferase"
/label=hRluc
/note="human codon-optimized"
/translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
CDS 4550..5341
/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="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV
LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS
HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG
LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL
ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4601..4620)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5211..5230
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5405..5453
/note="synthetic polyadenylation signal"
promoter 5609..5713
/gene="bla"
/label=AmpR promoter
CDS 5714..6574
/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 complement(5932..5951)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin complement(6732..7320)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6812..6831)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
misc_feature 7436..7719
/label=cer region
/note="ColE1-derived recombination site that helps to
maintain plasmids as monomers"
promoter 7776..156
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(7811..7834)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
ORIGIN
1 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
61 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
121 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag cttggcaatc
181 cggtactgtt ggtaaagcca ccatggaaga tgccaaaaac attaagaagg gcccagcgcc
241 attctaccca ctcgaagacg ggaccgccgg cgagcagctg cacaaagcca tgaagcgcta
301 cgccctggtg cccggcacca tcgcctttac cgacgcacat atcgaggtgg acattaccta
361 cgccgagtac ttcgagatga gcgttcggct ggcagaagct atgaagcgct atgggctgaa
421 tacaaaccat cggatcgtgg tgtgcagcga gaatagcttg cagttcttca tgcccgtgtt
481 gggtgccctg ttcatcggtg tggctgtggc cccagctaac gacatctaca acgagcgcga
541 gctgctgaac agcatgggca tcagccagcc caccgtcgta ttcgtgagca agaaagggct
601 gcaaaagatc ctcaacgtgc aaaagaagct accgatcata caaaagatca tcatcatgga
661 tagcaagacc gactaccagg gcttccaaag catgtacacc ttcgtgactt cccatttgcc
721 acccggcttc aacgagtacg acttcgtgcc cgagagcttc gaccgggaca aaaccatcgc
781 cctgatcatg aacagtagtg gcagtaccgg attgcccaag ggcgtagccc taccgcaccg
841 caccgcttgt gtccgattca gtcatgcccg cgaccccatc ttcggcaacc agatcatccc
901 cgacaccgct atcctcagcg tggtgccatt tcaccacggc ttcggcatgt tcaccacgct
961 gggctacttg atctgcggct ttcgggtcgt gctcatgtac cgcttcgagg aggagctatt
1021 cttgcgcagc ttgcaagact ataagattca atctgccctg ctggtgccca cactatttag
1081 cttcttcgct aagagcactc tcatcgacaa gtacgaccta agcaacttgc acgagatcgc
1141 cagcggcggg gcgccgctca gcaaggaggt aggtgaggcc gtggccaaac gcttccacct
1201 accaggcatc cgccagggct acggcctgac agaaacaacc agcgccattc tgatcacccc
1261 cgaaggggac gacaagcctg gcgcagtagg caaggtggtg cccttcttcg aggctaaggt
1321 ggtggacttg gacaccggta agacactggg tgtgaaccag cgcggcgagc tgtgcgtccg
1381 tggccccatg atcatgagcg gctacgttaa caaccccgag gctacaaacg ctctcatcga
1441 caaggacggc tggctgcaca gcggcgacat cgcctactgg gacgaggacg agcacttctt
1501 catcgtggac cggctgaaga gcctgatcaa atacaagggc taccaggtag ccccagccga
1561 actggagagc atcctgctgc aacaccccaa catcttcgac gccggggtcg ccggcctgcc
1621 cgacgacgat gccggcgagc tgcccgccgc agtcgtcgtg ctggaacacg gtaaaaccat
1681 gaccgagaag gagatcgtgg actatgtggc cagccaggtt acaaccgcca agaagctgcg
1741 cggtggtgtt gtgttcgtgg acgaggtgcc taaaggactg accggcaagt tggacgcccg
1801 caagatccgc gagattctca ttaaggccaa gaagggcggc aagatcgccg tgtaattcta
1861 gttgtttaaa cgagctcgct agcggaggac gaacatccaa ccttcccaaa cgcctcccct
1921 gccccaatcc ctttattacc ccctccttca gacaccctca acctcttctg gctcaaaaag
1981 agaattgggg gcttagggtc ggaacccaag cttagaactt taagcaacaa gaccaccact
2041 tcgaaacctg ggattcagga atgtgtggcc tgcacagtga agtgctggca accactaaga
2101 attcaaactg gggcctccag aactcactgg ggcctacagc tttgatccct gacatctgga
2161 atctggagac cagggagcct ttggttctgg ccagaatgct gcaggacttg agaagacctc
2221 acctagaaat tgacacaagt ggaccttagg ccttcctctc tccagatgtt tccagacttc
2281 cttgagacac ggagcccagc cctccccatg gagccagctc cctctattta tgtttgcact
2341 tgtgattatt tattatttat ttattattta tttatttaca gatgaatgta tttatttggg
2401 agaccggggt atcctggggg acccaatgta ggagctgcct tggctcagac atgttttccg
2461 tgaaaacgga gctgaacaat aggctgttcc catgtagccc cctggcctct gtgccttctt
2521 ttgattatgt tttttaaaat atttatctga ttaagttgtc taaacaatgc tgatttggtg
2581 accaactgtc actcattgct gagcctctgc tccccagggg agttgtgtct gtaatcgccc
2641 tactattcag tggcgagatc tagagtcgac ctgcaggcat gcaagctgat ccggctgcta
2701 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
2761 cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
2821 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
2881 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
2941 tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt
3001 ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
3061 cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg
3121 aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt
3181 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
3241 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
3301 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
3361 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
3421 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
3481 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tgattcttct gacacaacag
3541 tctcgaacca aaggctggag ccaccatggc ttccaaggtg tacgaccccg agcaacgcaa
3601 acgcatgatc actgggcctc agtggtgggc tcgctgcaag caaatgaacg tgctggactc
3661 cttcatcaac tactatgatt ccgagaagca cgccgagaac gccgtgattt ttctgcatgg
3721 taacgctgcc tccagctacc tgtggaggca cgtcgtgcct cacatcgagc ccgtggctag
3781 atgcatcatc cctgatctga tcggaatggg taagtccggc aagagcggga atggctcata
3841 tcgcctcctg gatcactaca agtacctcac cgcttggttc gagctgctga accttccaaa
3901 gaaaatcatc tttgtgggcc acgactgggg ggcttgtctg gcctttcact actcctacga
3961 gcaccaagac aagatcaagg ccatcgtcca tgctgagagt gtcgtggacg tgatcgagtc
4021 ctgggacgag tggcctgaca tcgaggagga tatcgccctg atcaagagcg aagagggcga
4081 gaaaatggtg cttgagaata acttcttcgt cgagaccatg ctcccaagca agatcatgcg
4141 gaaactggag cctgaggagt tcgctgccta cctggagcca ttcaaggaga agggcgaggt
4201 tagacggcct accctctcct ggcctcgcga gatccctctc gttaagggag gcaagcccga
4261 cgtcgtccag attgtccgca actacaacgc ctaccttcgg gccagcgacg atctgcctaa
4321 gatgttcatc gagtccgacc ctgggttctt ttccaacgct attgtcgagg gagctaagaa
4381 gttccctaac accgagttcg tgaaggtgaa gggcctccac ttcagccagg aggacgctcc
4441 agatgaaatg ggtaagtaca tcaagagctt cgtggagcgc gtgctgaaga acgagcagac
4501 cggtggtggg agcggaggtg gcggatcagg tggcggaggc tccggaggga ttgaacaaga
4561 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
4621 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
4681 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
4741 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
4801 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
4861 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
4921 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
4981 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
5041 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
5101 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
5161 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
5221 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
5281 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
5341 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg
5401 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat
5461 agcgataagg atcctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac
5521 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg tttcttagac
5581 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
5641 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
5701 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
5761 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
5821 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
5881 gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc tgctatgtgg
5941 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
6001 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
6061 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
6121 tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
6181 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
6241 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
6301 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
6361 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
6421 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
6481 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
6541 tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc aagcgacgcc
6601 caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
6661 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
6721 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
6781 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
6841 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
6901 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
6961 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
7021 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
7081 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
7141 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
7201 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
7261 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa
7321 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
7381 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt atagtccgga
7441 aatacaggaa cgcacgctgg atggcccttc gctgggatgg tgaaaccatg aaaaatggca
7501 gcttcagtgg attaagtggg ggtaatgtgg cctgtaccct ctggttgcat aggtattcat
7561 acggttaaaa tttatcaggc gcgattgcgg cagtttttcg ggtggtttgt tgccattttt
7621 acctgtctgc tgccgtgatc gcgctgaacg cgttttagcg gtgcgtacaa ttaagggatt
7681 atggtaaatc cacttactgt ctgccctcgt agccatcgag ataaaccgca gtactccggc
7741 cacgatgcgt ccggcgtaga ggatcgagat ctaccgggta ggggaggcgc ttttcccaag
7801 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
7861 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
7921 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
7981 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
8041 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
8101 ctccttcgct ttctgggct
//
