| 质粒名称: | PG13-luc (wt p53 binding sites) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 16442 |
| 存储实验室: | Bert Vogelstein |
|
载体骨架基本信息 |
|
| 载体名称: | pBluescript II SK + luc |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5800 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: | p53-binding site |
| 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（C端在主干上） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII（如果被摧毁则未知） |
| 3' 克隆位点: | EcoRI（如果被摧毁则未知） |
| 5' 测序引物 | T7 |
| 3' 测序引物 | LucNrev |
|
文献引用方法 |
|
| 材料和方法部分: | PG13-luc (wt p53 binding sites) was a gift from Bert Vogelstein (Addgene plasmid # 16442 ; http://n2t.net/addgene:16442 ; RRID:Addgene_16442) |
| 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 PG13-luc (wt p53 binding sites) 6038 bp ds-DNA circular SYN 12-APR-2019
DEFINITION Luciferase reporter containing 13 copies of the p53-binding
consensus sequence.
ACCESSION .
VERSION .
KEYWORDS PG13-luc (wt p53 binding sites)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6038)
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 6038)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 12, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6038
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(421..437)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 491..2143
/codon_start=1
/gene="luc"
/product="firefly luciferase"
/label=luciferase
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
primer_bind complement(557..575)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
intron 2274..2339
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 2469..2489
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 2914..3048
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2951..2970)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3005..3024
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3045..3061)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3098..3116)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3137..3153)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3137..3153)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3150..3172)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3161..3177
/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(3185..3215)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3230..3251
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3368..3385)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3539..4127)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3619..3638)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4298..5158)
/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 4921..4940
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5159..5263)
/gene="bla"
/label=AmpR promoter
rep_origin complement(5289..5744)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5426..5447)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5638..5657
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5871..5893
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5885..5902
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 5886..5902
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5909..5928
/label=T7
/note="T7 promoter, forward primer"
promoter 5909..5927
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 5952..5968
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 5953..5969
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 ccaggcaagt ccaggcaggc caggcaagtc caggcaggcc aggcaagtcc aggcaggcca
61 ggcaagtcca ggcaggccag gcaagtccag gcaggccagg caagtccagg caggccaggc
121 aagtccaggc agggccaggc aagtccaggc aggccaggca agtccaggca ggccaggcaa
181 gtccaggcag gatcgaattc gatctgtttt ttttagtatt aagcagaggc cgggggcccc
241 tggcctccgc ttactctgga gaaaaagaag agaggcattc cagaggcaac ttgtcaaaac
301 aggactggcg ccttggaggc gctgtggggc cacccaaatt gatataatta agccccaacc
361 gcctcttccc gcctcatttc agcctcacca ccctcgacaa gcttatcgat accgtcgggg
421 gatccactag ttctagagcg gccgccaccg cggtggagct cgagcttggc attccggtac
481 tgttggtaaa atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatcctct
541 agaggatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
601 tggaacaatt gcttttacag atgcacatat cgaggtgaac atcacgtacg cggaatactt
661 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
721 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
781 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
841 tatgaacatt tcgcagccta ccgtagtgtt tgtttccaaa aaggggttgc aaaaaatttt
901 gaacgtgcaa aaaaaattac caataatcca gaaaattatt atcatggatt ctaaaacgga
961 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
1021 tgaatacgat tttgtaccag agtcctttga tcgtgacaaa acaattgcac tgataatgaa
1081 ttcctctgga tctactgggt tacctaaggg tgtggccctt ccgcatagaa ctgcctgcgt
1141 cagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
1201 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
1261 atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttt tacgatccct
1321 tcaggattac aaaattcaaa gtgcgttgct agtaccaacc ctattttcat tcttcgccaa
1381 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctgggggcgc
1441 acctctttcg aaagaagtcg gggaagcggt tgcaaaacgc ttccatcttc cagggatacg
1501 acaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
1561 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
1621 taccgggaaa acgctgggcg ttaatcagag aggcgaatta tgtgtcagag gacctatgat
1681 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
1741 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tagttgaccg
1801 cttgaagtct ttaattaaat acaaaggata tcaggtggcc cccgctgaat tggaatcgat
1861 attgttacaa caccccaaca tcttcgacgc gggcgtggca ggtcttcccg acgatgacgc
1921 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
1981 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
2041 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
2101 gatcctcata aaggccaaga agggcggaaa gtccaaattg taaaatgtaa ctgtattcag
2161 cgatgacgaa attcttagct attgtaatac tctagaggat ctttgtgaag gaaccttact
2221 tctgtggtgt gacataattg gacaaactac ctacagagat ttaaagctct aaggtaaata
2281 taaaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt gtattttaga
2341 ttccaaccta tggaactgat gaatgggagc agtggtggaa tgcctttaat gaggaaaacc
2401 tgttttgctc agaagaaatg ccatctagtg atgatgaggc tactgctgac tctcaacatt
2461 ctactcctcc aaaaaagaag agaaaggtag aagaccccaa ggactttcct tcagaattgc
2521 taagtttttt gagtcatgct gtgtttagta atagaactct tgcttgcttt gctatttaca
2581 ccacaaagga aaaagctgca ctgctataca agaaaattat ggaaaaatat tctgtaacct
2641 ttataagtag gcataacagt tataatcata acatactgtt ttttcttact ccacacaggc
2701 atagagtgtc tgctattaat aactatgctc aaaaattgtg tacctttagc tttttaattt
2761 gtaaaggggt taataaggaa tatttgatgt atagtgcctt gactagagat cataatcagc
2821 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
2881 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
2941 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
3001 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcca ctagttctag
3061 agcggccgcc accgcggtgg agctccagct tttgttccct ttagtgaggg ttaatttcga
3121 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
3181 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
3241 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
3301 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
3361 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
3421 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
3481 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
3541 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
3601 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
3661 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
3721 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
3781 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
3841 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
3901 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
3961 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
4021 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
4081 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
4141 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
4201 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
4261 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
4321 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
4381 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
4441 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
4501 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
4561 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
4621 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
4681 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
4741 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
4801 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
4861 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
4921 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
4981 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
5041 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
5101 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
5161 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
5221 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
5281 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
5341 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
5401 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
5461 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
5521 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
5581 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
5641 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
5701 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
5761 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
5821 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
5881 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattgggtac
5941 cgggcccccc ctcgaggtcg acggtatcga taagcttgat gccaggcaag tccaggcagg
6001 ccaggcaagt ccaggcaggc caggcaagtc caggcagg
//
