| 质粒名称: | pGL3-Basic-IRES |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 64784 |
| 存储实验室: | Joshua Mendell |
|
载体骨架基本信息 |
|
| 载体名称: | pGL3-Basic |
| 载体出品公司: | Promega |
| 空载体大小: | 4818 bp |
| 完整质粒大小: | 5408 bp |
| 载体类型: | Luciferase |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 荧光素酶 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | RVprimer3 |
| 3’测序引物 | GLprimer2 |
|
文献引用方法 |
|
| 材料和方法部分: | pGL3-Basic-IRES was a gift from Joshua Mendell (Addgene plasmid # 64784 ; http://n2t.net/addgene:64784 ; RRID:Addgene_64784) |
| 参考文献部分: | Transactivation of miR-34a by p53 broadly influences gene expression and promotes apoptosis. Chang TC, Wentzel EA, Kent OA, Ramachandran K, Mullendore M, Lee KH, Feldmann G, Yamakuchi M, Ferlito M, Lowenstein CJ, Arking DE, Beer MA, Maitra A, Mendell JT. Mol Cell. 2007 Jun 8;26(5):745-52. Epub 2007 May 31. 10.1016/j.molcel.2007.05.010 PubMed 17540599 |
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 pGL3-Basic-IRES 5408 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Luciferase reporter vector.
ACCESSION .
VERSION .
KEYWORDS pGL3-Basic-IRES
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5408)
AUTHORS Chang TC, Wentzel EA, Kent OA, Ramachandran K, Mullendore M, Lee KH,
Feldmann G, Yamakuchi M, Ferlito M, Lowenstein CJ, Arking DE, Beer
MA, Maitra A, Mendell JT
TITLE Transactivation of miR-34a by p53 broadly influences gene expression
and promotes apoptosis.
JOURNAL Mol Cell. 2007 Jun 8;26(5):745-52. Epub 2007 May 31.
PUBMED 17540599
REFERENCE 2 (bases 1 to 5408)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5408
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 164..626
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(220..237)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 447..466
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
regulatory 672..681
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 678..2330
/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(744..762)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
polyA_signal 2371..2492
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2382..2401)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2436..2455
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2740..2757)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(2911..3499)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2991..3010)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3670..4530)
/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 4293..4312
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4531..4635)
/gene="bla"
/label=AmpR promoter
rep_origin 4662..5117
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4749..4768)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4959..4980
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 5248..5296
/note="synthetic polyadenylation signal"
misc_feature 5310..5401
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind 5350..5369
/label=RVprimer3
/note="pGL3 vector, forward primer"
ORIGIN
1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgaccccatg catcgaattc
61 cgcccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
121 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
181 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
241 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
301 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
361 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
421 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
481 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
541 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
601 ttttcctttg aaaaacacga tgataagatc tgcgatctaa gtaagcttgg cattccggta
661 ctgttggtaa agccaccatg gaagacgcca aaaacataaa gaaaggcccg gcgccattct
721 atccgctgga agatggaacc gctggagagc aactgcataa ggctatgaag agatacgccc
781 tggttcctgg aacaattgct tttacagatg cacatatcga ggtggacatc acttacgctg
841 agtacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg ctgaatacaa
901 atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg
961 cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc
1021 tcaacagtat gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa
1081 aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc atggattcta
1141 aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat ctacctcccg
1201 gttttaatga atacgatttt gtgccagagt ccttcgatag ggacaagaca attgcactga
1261 tcatgaactc ctctggatct actggtctgc ctaaaggtgt cgctctgcct catagaactg
1321 cctgcgtgag attctcgcat gccagagatc ctatttttgg caatcaaatc attccggata
1381 ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact acactcggat
1441 atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga
1501 ggagccttca ggattacaag attcaaagtg cgctgctggt gccaacccta ttctccttct
1561 tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa attgcttctg
1621 gtggcgctcc cctctctaag gaagtcgggg aagcggttgc caagaggttc catctgccag
1681 gtatcaggca aggatatggg ctcactgaga ctacatcagc tattctgatt acacccgagg
1741 gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg
1801 atctggatac cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc
1861 ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg attgacaagg
1921 atggatggct acattctgga gacatagctt actgggacga agacgaacac ttcttcatcg
1981 ttgaccgcct gaagtctctg attaagtaca aaggctatca ggtggctccc gctgaattgg
2041 aatccatctt gctccaacac cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg
2101 atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag acgatgacgg
2161 aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag
2221 gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa
2281 tcagagagat cctcataaag gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc
2341 ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
2401 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
2461 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
2521 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
2581 gtaaaatcga taaggatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
2641 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
2701 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
2761 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
2821 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
2881 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
2941 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
3001 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
3061 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
3121 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
3181 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
3241 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
3301 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
3361 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
3421 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
3481 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
3541 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
3601 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
3661 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
3721 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
3781 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
3841 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
3901 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
3961 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
4021 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
4081 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
4141 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
4201 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
4261 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
4321 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
4381 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
4441 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
4501 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
4561 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
4621 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc
4681 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
4741 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
4801 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
4861 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
4921 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
4981 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
5041 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
5101 attaacgctt acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
5161 ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa
5221 ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt
5281 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
5341 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
5401 tatcgata
//
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