| 质粒名称: | WWP-Luc (p21/WAF1 promoter) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 16451 |
| 存储实验室: | Bert Vogelstein |
|
载体骨架基本信息 |
|
| 载体名称: | pBluescript II KS (+) + luc |
| 载体出品公司: | -- |
| 载体抗性: | 氯霉素 |
| 空载体大小: | 5600 bp |
| 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: | WAF1 promoter,p21 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2400 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | luciferase (主链C端) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII（销毁后未知） |
| 3' 克隆位点: | HindIII（销毁后未知） |
| 5' 测序引物 | T7 |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | WWP-Luc (p21/WAF1 promoter) was a gift from Bert Vogelstein (Addgene plasmid # 16451 ; http://n2t.net/addgene:16451 ; RRID:Addgene_16451) |
| 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 WWP-Luc (p21/WAF1 promoter) 8366 bp ds-DNA circular SYN 29-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS WWP-Luc (p21/WAF1 promoter)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8366)
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 8366)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 29, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8366
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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)."
primer_bind 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 240..260
/label=T3
/note="T3 promoter, forward primer"
promoter 242..260
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
polyA_signal 291..425
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(315..334)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 369..388
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(850..870)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
intron 1000..1065
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS complement(1196..2848)
/codon_start=1
/gene="luc"
/product="firefly luciferase"
/label=luciferase
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
primer_bind 2764..2782
/label=LucNrev
/note="Firefly luciferase, reverse primer"
primer_bind 2902..2918
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2917..2933
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 2918..2934
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5306..5322)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5354..5373)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5355..5373)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5383..5400)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5383..5399)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5392..5414)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 5541..5996
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5628..5647)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5838..5859
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 6022..6126
/gene="bla"
/label=AmpR promoter
promoter 6446..6548
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 6549..7208
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(6615..6634)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
rep_origin 7597..8185
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8086..8105
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8339..8356
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
241 caattaaccc tcactaaagg gaacaaaagc tgggtaccgg gccccccctc gatccagaca
301 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
361 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
421 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
481 ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat ctctagtcaa
541 ggcactatac atcaaatatt ccttattaac ccctttacaa attaaaaagc taaaggtaca
601 caatttttga gcatagttat taatagcaga cactctatgc ctgtgtggag taagaaaaaa
661 cagtatgtta tgattataac tgttatgcct acttataaag gttacagaat atttttccat
721 aattttcttg tatagcagtg cagctttttc ctttgtggtg taaatagcaa agcaagcaag
781 agttctatta ctaaacacag catgactcaa aaaacttagc aattctgaag gaaagtcctt
841 ggggtcttct acctttctct tcttttttgg aggagtagaa tgttgagagt cagcagtagc
901 ctcatcatca ctagatggca tttcttctga gcaaaacagg ttttcctcat taaaggcatt
961 ccaccactgc tcccattcat cagttccata ggttggaatc taaaatacac aaacaattag
1021 aatcagtagt ttaacacatt atacacttaa aaattttata tttaccttag agctttaaat
1081 ctctgtaggt agtttgtcca attatgtcac accacagaag taaggttcct tcacaaagat
1141 cctctagagt attacaatag ctaagaattt cgtcatcgct gaatacagtt acattttaca
1201 atttggactt tccgcccttc ttggccttta tgaggatctc tctgattttt cttgcgtcga
1261 gttttccggt aagacctttc ggtacttcgt ccacaaacac aactcctccg cgcaactttt
1321 tcgcggttgt tacttgactg gcgacgtaat ccacgatctc tttttccgtc atcgtctttc
1381 cgtgctccaa aacaacaacg gcggcgggaa gttcaccggc gtcatcgtcg ggaagacctg
1441 ccacgcccgc gtcgaagatg ttggggtgtt gtaacaatat cgattccaat tcagcggggg
1501 ccacctgata tcctttgtat ttaattaaag acttcaagcg gtcaactatg aagaagtgtt
1561 cgtcttcgtc ccagtaagct atgtctccag aatgtagcca tccatccttg tcaatcaagg
1621 cgttggtcgc ttccggattg tttacataac cggacataat cataggtcct ctgacacata
1681 attcgcctct ctgattaacg cccagcgttt tcccggtatc cagatccaca accttcgctt
1741 caaaaaatgg aacaacttta ccgaccgcgc ccggtttatc atccccctcg ggtgtaatca
1801 gaatagctga tgtagtctca gtgagcccat atccttgtcg tatccctgga agatggaagc
1861 gttttgcaac cgcttccccg acttctttcg aaagaggtgc gcccccagaa gcaatttcgt
1921 gtaaattaga taaatcgtat ttgtcaatca gagtgctttt ggcgaagaat gaaaataggg
1981 ttggtactag caacgcactt tgaattttgt aatcctgaag ggatcgtaaa aacagctctt
2041 cttcaaatct atacattaag acgactcgaa atccacatat caaatatccg agtgtagtaa
2101 acattccaaa accgtgatgg aatggaacaa cacttaaaat cgcagtatcc ggaatgattt
2161 gattgccaaa aataggatct ctggcatgcg agaatctgac gcaggcagtt ctatgcggaa
2221 gggccacacc cttaggtaac ccagtagatc cagaggaatt cattatcagt gcaattgttt
2281 tgtcacgatc aaaggactct ggtacaaaat cgtattcatt aaaaccggga ggtagatgag
2341 atgtgacgaa cgtgtacatc gactgaaatc cctggtaatc cgttttagaa tccatgataa
2401 taattttctg gattattggt aatttttttt gcacgttcaa aattttttgc aacccctttt
2461 tggaaacaaa cactacggta ggctgcgaaa tgttcatact gttgagcaat tcacgttcat
2521 tataaatgtc gttcgcgggc gcaactgcaa ctccgataaa taacgcgccc aacaccggca
2581 taaagaattg aagagagttt tcactgcata cgacgattct gtgatttgta ttcagcccat
2641 atcgtttcat agcttctgcc aaccgaacgg acatttcgaa gtattccgcg tacgtgatgt
2701 tcacctcgat atgtgcatct gtaaaagcaa ttgttccagg aaccagggcg tatctcttca
2761 tagccttatg cagttgctct ccagcggttc catcctctag aggatagaat ggcgccgggc
2821 ctttctttat gtttttggcg tcttccattt taccaacagt accggaatgc caagctcgag
2881 ctccaccgcg gtggcggccg ctctagaact agtggatcga ggtcgacggt atcgataagc
2941 ttctgacttc ggcagctgct cacacctcag ctggcgcagc tctgcgcggc cctgatatac
3001 aaccgccccg cccgggcccg cctcaaggag gcgggacccg cgctcggccc accgcgccgc
3061 ccgggaccgc cctccccggg gcccaggctc ctggctgccc agcgcggagc cagctgagcc
3121 tggccgagtt ccagcaggcc agccggtccc ggaacctcgc gtgctgcaga ggggcacttc
3181 cctcctcccc cagtccctcg cctgcgttgg tgcgctggac acatttcccc acgaagtgag
3241 ccacaaatct ggcttttttt acttggagaa tgagttggca ctctccagga ggacacagca
3301 ctgttagaat gagccccctt tctggctcac cgctgaccca ctctggcagg caaggattta
3361 cccaatgcag ctgaaaagat caggaggatg acattaatac ataaaaattc ataaattata
3421 aaaacgatgc acctctctgc aatttccaga aaagccccac aatatcacct ctattcccac
3481 tgatccctca ctaggtcacc tctcccagaa gcacctggag cacctagaca ccccaacaaa
3541 gcatcttgag gccggaatga tttttcagtc ccgtttattt cacagatgag gaaattgagg
3601 tccactgaac ttaagtatac aaagttgttg attgtcacat gcttccggga aggagggaat
3661 tggagagact accaaaaaag ggcaacctga tctccaggga aacagaagaa ttggacattg
3721 aaccaatctc cctacaccct acactcacct gaacagaaga aacccctgtg gttgcagcag
3781 ctttgttggc caggaagggg aggatttgac gagtgagttg tctgtctcct gaatactccc
3841 cacatagccc gtatacactg ctggggaaac tggggctcag agaagtctgg tgacctaact
3901 cagatcatgc agcaaagaaa tgactatagt tggaacacag gacttttgcc tcctgcccgg
3961 ggctctctgc ttgtcatcct ttatttctgt ggttccaaaa tgacaaaaat gccaaataac
4021 cctcatttgc agatggttta tggagatgac gtaaataaag gacaattctg gaagtgtcta
4081 ctggttcttc tgacagacac agaaatgagt gatgtgccta tccgctccca tctacctcac
4141 acccctgacc tcccctggac ttcacctttg cctcctttct gtgcctgaaa catttgcagt
4201 tttgctttta aaaaattgca gaggatggat tgttcatctg aacagaaatc ccactgaaaa
4261 acagaaccca ggcttggagc agctacaatt actgacatct caggctgctc agagtctgga
4321 aatctctgcc cagacatgcc cagtcttctt cctctaacgc agctgacctc ggggaggaca
4381 ggcttctttc tcctctgctg tggggatggg agcccaggta ggggtgggag gacacagcag
4441 caggccccct tggcctgcct cgccctggag tcaggccagg attgtggcta aaccccagaa
4501 gggccacagc acagcctcag gaccccactc taagccccac tgctgtctgc accttcgctc
4561 ctatacatac aagccatcca aagggctggt tgtcaaatgt ccagcagagg acaggcacct
4621 tctcccactg ccttgaagcc cctctgcttt caggcatttc aaatagacta gacattgtct
4681 gtctgtttac ttcggggaga aatttttttt taatgaaggc ttgtaatgaa ttatttaaat
4741 ttttgtggta ctgagcttca caatgaaata attctttttg ttgttgtttt tgggaaagaa
4801 ttttttttat aacttctagc tcaccaccac cacgacattc aatatttctt aagtacctac
4861 taagtgtcta gtactattca gtgctttaaa aaaaaaagtt tgcaaccatg cacttgaatg
4921 tgtacccaga cacactctaa gggaggactt ctgccctgag gaaggcaagg gagcaggctg
4981 ttaaaagtca cacacagaat ctgactccca gcacacactc acacaagcac acatgcatca
5041 gatcccagcc ctgtcgcaag gatctgctgg cagatcacat accctgttca gagtaacagg
5101 ctaaggctta cctggggcct ttagaggtct tctgtctcct accatcccct tcctcacctg
5161 ataacaggca gcccaaggac aaaatagcca ccagcctctt ctatgccaga gctcaacatg
5221 ttgggacatg ttcctaatgg ccagaaagcc aatcagagcc acagcctgct gcccaagctt
5281 gatatcgaat tcctgcagcc cgggggatcc actagttcta gagcggccgc caccgcggtg
5341 gagctccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
5401 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
5461 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
5521 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
5581 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
5641 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
5701 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
5761 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
5821 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
5881 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
5941 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
6001 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
6061 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
6121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
6181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
6241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
6301 cgccccgaag aacgttttcc aatgatgagc acttttcgac cgaataaata cctgtgacgg
6361 aagatcactt cgcagaataa ataaatcctg gtgtccctgt tgataccggg aagccctggg
6421 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
6481 atgaaataag atcactaccg ggcgtatttt ttgagttgtc gagattttca ggagctaagg
6541 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
6601 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
6661 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
6721 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattacgt atggcaatga
6781 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
6841 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
6901 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
6961 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
7021 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
7081 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
7141 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
7201 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc tacgcctgaa
7261 taagtgataa taagcggatg aatggcagaa attcgaaagc aaattcgacc cggtcgtcgg
7321 ttcagggcag ggtcgttaaa tagccgctta tgtctattgc tggtttaccg gtttattgac
7381 taccggaagc agtgtgaccg tgtgcttctc aaatgcctga ggccagtttg ctcaggctct
7441 ccccgtggag gtaataattg acgatatgat cctttttttc tgatcaaaaa ggatctaggt
7501 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
7561 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
7621 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
7681 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
7741 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
7801 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
7861 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
7921 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
7981 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
8041 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
8101 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
8161 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
8221 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
8281 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
8341 cgagtcagtg agcgaggaag cggaag
//
