质粒名称: | pGL3 3 kb prom. CD274 |
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出品公司: | Addgene |
目录编号: | 107004 |
存储实验室: | Julian Downward |
载体骨架基本信息 |
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载体名称: | pGL3 |
载体出品公司: | Promega |
载体抗性: | 氨苄青霉素 |
载体类型: | 哺乳动物表达,荧光素酶 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | CD274,PD-L1, B7-H1 |
物种来源: | H. sapiens (human) |
插入大小: | 3073 bp |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | MluI(销毁后未知) |
3' 克隆位点: | XhoI(销毁后未知 |
5' 测序引物 | RVprimer3 |
3' 测序引物 | TCTTTATGTTTTTGGCGTC |
文献引用方法 |
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材料和方法部分: | pGL3 3 kb prom. CD274 was a gift from Julian Downward (Addgene plasmid # 107004 ; http://n2t.net/addgene:107004 ; RRID:Addgene_107004) |
参考文献部分: | Oncogenic RAS Signaling Promotes Tumor Immunoresistance by Stabilizing PD-L1 mRNA. Coelho MA, de Carne Trecesson S, Rana S, Zecchin D, Moore C, Molina-Arcas M, East P, Spencer-Dene B, Nye E, Barnouin K, Snijders AP, Lai WS, Blackshear PJ, Downward J. Immunity. 2017 Dec 19;47(6):1083-1099.e6. doi: 10.1016/j.immuni.2017.11.016. Epub 2017 Dec 12. 10.1016/j.immuni.2017.11.016 PubMed 29246442 |
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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 pGL3 3 kb prom. CD274 7879 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Modified Luciferase expression vector. ACCESSION . VERSION . KEYWORDS pGL3 3 kb prom. CD274 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7879) AUTHORS Coelho MA, de Carne Trecesson S, Rana S, Zecchin D, Moore C, Molina-Arcas M, East P, Spencer-Dene B, Nye E, Barnouin K, Snijders AP, Lai WS, Blackshear PJ, Downward J TITLE Oncogenic RAS Signaling Promotes Tumor Immunoresistance by Stabilizing PD-L1 mRNA. JOURNAL Immunity. 2017 Dec 19;47(6):1083-1099.e6. doi: 10.1016/j.immuni.2017.11.016. Epub 2017 Dec 12. PUBMED 29246442 REFERENCE 2 (bases 1 to 7879) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7879 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 7..55 /note="synthetic polyadenylation signal" misc_feature 69..160 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 109..128 /label=RVprimer3 /note="pGL3 vector, forward primer" regulatory 3310..3319 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3316..4968 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(3382..3400) /label=LucNrev /note="Firefly luciferase, reverse primer" polyA_signal 5009..5130 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5020..5039) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 5074..5093 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(5378..5395) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5549..6137) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5629..5648) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6308..7168) /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 6931..6950 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7169..7273) /gene="bla" /label=AmpR promoter rep_origin 7300..7755 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7387..7406) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7597..7618 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg 61 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag 121 gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgataggt accgagctct 181 tacgcgtctc cccagcttag gacaggaagg agagagacac agtgtgagcg agagaagcat 241 ctgattgtga gtgtcagatg aggacagctg ctactctctg gtcacccata catgttaatt 301 acccctgtca tttaaaccat tgttttgtgt aactagaaca aagtattctt tccacgaagt 361 ctgtttgaca cgagacacat ataggatgtg agtctatgac aagggtaggg gcaacaaatg 421 tgaggggttg gatggaaaca gaggaagaga aatgagcatt gcatagggca gtggtttaaa 481 accctacatg atcaaaaata gaacctggag acaaatcttg atattgtctt tatacataat 541 aaaatgcatt ttaaaatatt cgatgcaaac aaagcagttt gtcatgtctt ttgttttgag 601 tctctttagt atatgctgct tttgaagatc aagaacattt actggaaatt gctccttcac 661 caggaatttg ctcacatctc ttcaggtcca cttataagat cttgaaatca gtcctcctga 721 gatcagtaca aacgaggatc atctctcctg ctttagaagc aaaagtggca agcagggtag 781 acagaggcag aaggaaggat ggtactgata aatcagccaa attagatgga agaaagcgaa 841 ttacagctgt acttgcaaag aactcttggc tcattttttc ccttaaatat taaatcacta 901 gttttcaaaa cttcaaatgt gaacatgact cacctgagga caatgttaaa atgttgacct 961 ccaagaccct gcccccaagt ttggatccca ggcagaaccc tggaatctgt attttaaaca 1021 ttaaccccaa gtcgagtgta atgcaagtaa ccatcagaga cactctgaga aacagcacat 1081 tggcaatgat gaatttatat gtctaaaaat gaatagagta gatgttacat gaataggaag 1141 tggtggtatt caagatgacc atagtatcta gctcttcagc cctggctccc cactgctctt 1201 ctcccatctc agcacttctc catctatttc gtccaaataa aaaaattcac acagagtttc 1261 aggacttaac cccccactca ttaaccatct gttttgcttt acatattttt ctgaggtaat 1321 aaaatttctc tttttctaaa cacagcctgt ttttcaatct ccgggtagtt gatcaattgt 1381 atgggaaaat gaatggctga agggtagaaa caggtgggaa agatgaacaa aaacacgaat 1441 cctcacatta ctaatacgca aatcactgag cagcaagctg agcaaatacc ctcaattccc 1501 atcagcaact ttagagaaag gcaaattccg tttgcctcat tgatcattta ggtagaccct 1561 gaacactgct ttcataaaac aaaaacaaaa tacccatccc cagtttaaaa aattattcat 1621 agatcatcca ggccatctag gaggatatga tttaatcctg gctacttggt aaattatttg 1681 cccaagttaa ctcagctagt tagtggtaga tggctctgaa gccagttgtt tttttttgtt 1741 ttgttttttg cagacctcaa gagtcatgat gaactagcag atcataaagt ttatgccctg 1801 ggtcttgacc atttttagaa aaataaaaca ttaaatgaaa atatcagagg gcattgcaga 1861 tagtagatct aagtattttt tcatgaaact tgttgtacat gtgtgtgtca tacacagact 1921 atatatatgc agtacctgta aactgtattg ccacataatg tctatatttt cctagaggtc 1981 acagtcacca aagttgggaa gtcacccaac ttcgggaact ttgggaagtc acccaaactt 2041 acagtcacca aaattgctct attctactat gtgacctcaa aagtgatttg aaagaaggaa 2101 catctgagct gggcccaaac cctattgcaa ttttattggg gccaaagaga actccatgct 2161 cctgccaaat caaggcagtg tcagcctcaa taatttccca gataaaaata aaaatctgtg 2221 atacaatcag aatgtgaaaa ttcttatttt ggaagcaaat gtcataacca atgcaagggc 2281 tatctcaata ttcattcatt atgcagtatt ttgaactgca gttgaaatga ataagaagga 2341 aaggcaaaca acgaagagtc caatttctca atttagaaaa agagaaaaaa aagaaaaggg 2401 agcacacagg cacggtggct caagcctgta atatcagcac tttggcggat cacttgaggt 2461 caaggagttc gagaaaagag agcacctaga agttcagcgc gggataatac ttaagtaaat 2521 tatgacacca tcgtctgtca tcttgggccc attcactaac ccaaagcttt caaaagggct 2581 ttcttaaccc tcacctagaa taggcttccg cagccttaat ccttagggtg gcagaatatc 2641 agggaccctg agcattctta aaagatgtag ctcgggatgg gaagttcttt taatgacaaa 2701 gcaaatgaag tttcattatg tcgaggaact ttgaggaagt cacagaatcc acgatttaaa 2761 aatatatttc ctattataca cccatacaca cacacacaca cctactttct agaataaaaa 2821 ccaaagccat atgggtctgc tgctgacttt ttatatgttg tagagttata tcaagttatg 2881 tcaagatgtt cagtcacctt gaagaggctt ttatcagaaa gggggacgcc tttctgataa 2941 aggttaaggg gtaaccttaa gctcttaccc ctctgaaggt aaaatcaagg tgcgttcaga 3001 tgttggcttg ttgtaaattt ctttttttat taataacata ctaaatgtgg atttgcttta 3061 atcttcgaaa ctcttcccgg tgaaaatctc atttacaaga aaactggact gacatgtttc 3121 actttctgtt tcatttctat acacagcttt attcctagga caccaacact agatacctaa 3181 actgaaagct tccgccgatt tcaccgaagg tcaggaaagt ccaacgcccg gcaaactgga 3241 tttgctgcct tgggcagagc tcgagatctg cgatctaagt aagcttggca ttccggtact 3301 gttggtaaag ccaccatgga agacgccaaa aacataaaga aaggcccggc gccattctat 3361 ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg 3421 gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag 3481 tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat 3541 cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg 3601 ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc 3661 aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa 3721 attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa 3781 acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt 3841 tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc 3901 atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc 3961 tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact 4021 gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat 4081 ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg 4141 agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc 4201 gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt 4261 ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt 4321 atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac acccgagggg 4381 gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat 4441 ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct 4501 atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat 4561 ggatggctac attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt 4621 gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa 4681 tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat 4741 gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa 4801 aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga 4861 gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc 4921 agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaatt ctagagtcgg 4981 ggcggccggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca 5041 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt 5101 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt 5161 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt 5221 aaaatcgata aggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt 5281 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca 5341 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct 5401 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca 5461 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga 5521 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc 5581 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg 5641 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat 5701 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt 5761 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc 5821 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg 5881 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg 5941 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg 6001 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg 6061 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca 6121 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 6181 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 6241 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 6301 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 6361 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 6421 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 6481 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 6541 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 6601 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 6661 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 6721 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 6781 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 6841 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 6901 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 6961 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 7021 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 7081 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 7141 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 7201 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 7261 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat 7321 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag 7381 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 7441 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 7501 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt 7561 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 7621 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg 7681 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 7741 taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg 7801 tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg 7861 tacgggaggt acttggagc //