| 質粒名稱: | pGL3 3 kb prom. CD274 |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 107004 |
| 存儲實驗室: | Julian Downward |
|
載體骨架基本信息 |
|
| 載體名稱: | pGL3 |
| 載體出品公司: | Promega |
| 載體抗性: | 氨芐青霉素 |
| 載體類型: | 哺乳動物表達，熒光素酶 |
| 生長條件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名稱: | CD274,PD-L1, B7-H1 |
| 物種來源: | H. sapiens (human) |
| 插入大小: | 3073 bp |
| 克隆方法: | 限制性內切酶 |
| 5' 克隆位點: | MluI（銷毀后未知） |
| 3' 克隆位點: | XhoI（銷毀后未知 |
| 5' 測序引物 | RVprimer3 |
| 3' 測序引物 | TCTTTATGTTTTTGGCGTC |
|
文獻引用方法 |
|
| 材料和方法部分: | pGL3 3 kb prom. CD274 was a gift from Julian Downward (Addgene plasmid # 107004 ; //n2t.net/addgene:107004 ; RRID:Addgene_107004) |
| 參考文獻部分: | 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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pGL3 3 kb prom. CD274載體質粒基本信息
pGL3 3 kb prom. CD274質粒圖譜載體圖譜和pGL3 3 kb prom. CD274載體序列質粒序列多克隆位點信息
pGL3 3 kb prom. CD274質粒載體簡介
pGL3 3 kb prom. CD274質粒序列載體序列
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
//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
//