| 质粒名称: | MG15-luc (mut p53 binding sites) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 16443 |
| 存储实验室: | Bert Vogelstein |
|
载体骨架基本信息 |
|
| 载体名称: | -- |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 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: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | p53-binding site (mutant) |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | luciferase（主干上的C末端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRV（在克隆过程中被破坏） |
| 3' 克隆位点: | EcoRV（在克隆过程中被破坏） |
| 5' 测序引物 | M13pUC-Fwd |
| 3' 测序引物 | LucNRev |
|
文献引用方法 |
|
| 材料和方法部分: | MG15-luc (mut p53 binding sites) was a gift from Bert Vogelstein (Addgene plasmid # 16443 ; http://n2t.net/addgene:16443 ; RRID:Addgene_16443) |
| 参考文献部分: | WAF1, a potential mediator of p53 tumor suppression. el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM, Lin D, Mercer WE, Kinzler KW, Vogelstein B. Cell. 1993 Nov 19. 75(4):817-25. 10.1016/0092-8674(93)90500-P PubMed 8242752 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS MG15-luc (mut p53 binding sites) 5961 bp ds-DNA circular SYN 29-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS MG15-luc (mut p53 binding sites)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5961)
AUTHORS el-Deiry WS, Tokino T, Velculescu VE, Levy DB, Parsons R, Trent JM,
Lin D, Mercer WE, Kinzler KW, Vogelstein B
TITLE WAF1, a potential mediator of p53 tumor suppression.
JOURNAL Cell. 1993 Nov 19. 75(4):817-25.
PUBMED 8242752
REFERENCE 2 (bases 1 to 5961)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 29, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5961
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(410..426)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 480..2132
/codon_start=1
/gene="luc"
/product="firefly luciferase"
/label=luciferase
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
primer_bind complement(546..564)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
intron 2263..2328
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 2458..2478
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 2903..3037
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2940..2959)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2994..3013
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3034..3050)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3087..3105)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3126..3142)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3126..3142)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3139..3161)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3150..3166
/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(3174..3204)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3219..3240
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3357..3374)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3528..4116)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3608..3627)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4287..5147)
/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 4910..4929
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5148..5252)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5278..5733)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5415..5436)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5627..5646
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5860..5882
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5874..5891
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 5875..5891
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5898..5917
/label=T7
/note="T7 promoter, forward primer"
promoter 5898..5916
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 5941..5957
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 5942..5958
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 agcttgatcc ttaatggact ttaatggcct tatggacttt aatggcctta atggacttta
61 atggccttaa tggactttaa tggccttaat ggactttaat ggccttaatg gactttaatg
121 gccttaatgg actttaatgg ccttaatgga ctttaatggc cttaatggac tttaatggcc
181 atcgaattcg atctgttttt tttagtatta agcagaggcc gggggcccct ggcctccgct
241 tactctggag aaaaagaaga gaggcattcc agaggcaact tgtcaaaaca ggactggcgc
301 cttggaggcg ctgtggggcc acccaaattg atataattaa gccccaaccg cctcttcccg
361 cctcatttca gcctcaccac cctcgacaag cttatcgata ccgtcggggg atccactagt
421 tctagagcgg ccgccaccgc ggtggagctc gagcttggca ttccggtact gttggtaaaa
481 tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatcctcta gaggatggaa
541 ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg
601 cttttacaga tgcacatatc gaggtgaaca tcacgtacgc ggaatacttc gaaatgtccg
661 ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat
721 gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg
781 cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgaacattt
841 cgcagcctac cgtagtgttt gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa
901 aaaaattacc aataatccag aaaattatta tcatggattc taaaacggat taccagggat
961 ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt
1021 ttgtaccaga gtcctttgat cgtgacaaaa caattgcact gataatgaat tcctctggat
1081 ctactgggtt acctaagggt gtggcccttc cgcatagaac tgcctgcgtc agattctcgc
1141 atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg
1201 ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc
1261 gagtcgtctt aatgtataga tttgaagaag agctgttttt acgatccctt caggattaca
1321 aaattcaaag tgcgttgcta gtaccaaccc tattttcatt cttcgccaaa agcactctga
1381 ttgacaaata cgatttatct aatttacacg aaattgcttc tgggggcgca cctctttcga
1441 aagaagtcgg ggaagcggtt gcaaaacgct tccatcttcc agggatacga caaggatatg
1501 ggctcactga gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg
1561 cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa
1621 cgctgggcgt taatcagaga ggcgaattat gtgtcagagg acctatgatt atgtccggtt
1681 atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg
1741 gagacatagc ttactgggac gaagacgaac acttcttcat agttgaccgc ttgaagtctt
1801 taattaaata caaaggatat caggtggccc ccgctgaatt ggaatcgata ttgttacaac
1861 accccaacat cttcgacgcg ggcgtggcag gtcttcccga cgatgacgcc ggtgaacttc
1921 ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt
1981 acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg
2041 aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa
2101 aggccaagaa gggcggaaag tccaaattgt aaaatgtaac tgtattcagc gatgacgaaa
2161 ttcttagcta ttgtaatact ctagaggatc tttgtgaagg aaccttactt ctgtggtgtg
2221 acataattgg acaaactacc tacagagatt taaagctcta aggtaaatat aaaattttta
2281 agtgtataat gtgttaaact actgattcta attgtttgtg tattttagat tccaacctat
2341 ggaactgatg aatgggagca gtggtggaat gcctttaatg aggaaaacct gttttgctca
2401 gaagaaatgc catctagtga tgatgaggct actgctgact ctcaacattc tactcctcca
2461 aaaaagaaga gaaaggtaga agaccccaag gactttcctt cagaattgct aagttttttg
2521 agtcatgctg tgtttagtaa tagaactctt gcttgctttg ctatttacac cacaaaggaa
2581 aaagctgcac tgctatacaa gaaaattatg gaaaaatatt ctgtaacctt tataagtagg
2641 cataacagtt ataatcataa catactgttt tttcttactc cacacaggca tagagtgtct
2701 gctattaata actatgctca aaaattgtgt acctttagct ttttaatttg taaaggggtt
2761 aataaggaat atttgatgta tagtgccttg actagagatc ataatcagcc ataccacatt
2821 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
2881 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
2941 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
3001 gtccaaactc atcaatgtat cttatcatgt ctggatccac tagttctaga gcggccgcca
3061 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa
3121 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
3181 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
3241 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
3301 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
3361 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
3421 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
3481 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
3541 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
3601 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
3661 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
3721 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
3781 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
3841 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
3901 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
3961 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
4021 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
4081 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
4141 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
4201 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
4261 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
4321 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
4381 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
4441 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
4501 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
4561 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
4621 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
4681 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
4741 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
4801 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
4861 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
4921 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
4981 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
5041 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
5101 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
5161 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
5221 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
5281 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
5341 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
5401 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
5461 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
5521 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
5581 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
5641 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
5701 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
5761 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
5821 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
5881 cgacggccag tgaattgtaa tacgactcac tatagggcga attgggtacc gggccccccc
5941 tcgaggtcga cggtatcgat a
//
