| 质粒名称: | p53-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 11770 |
| 存储实验室: | Geoff Wahl |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4733 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | p53 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | 72R variant of p53 |
| GenBank编号 | AAD28628.1 |
| 启动子 | -- |
| 标签/融合蛋白: | GFP（主干上的C终端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | SalI (未破坏) |
| 3' 克隆位点: | BamHI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | EGFP-N |
|
文献引用方法 |
|
| 材料和方法部分: | p53-GFP was a gift from Geoff Wahl (Addgene plasmid # 11770 ; http://n2t.net/addgene:11770 ; RRID:Addgene_11770) |
买家导航
p53GFP载体基本信息
p53GFP载体图谱质粒图谱和多克隆位点信息
p53GFP载体简介
p53GFP载体序列
LOCUS p53-GFP 5906 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS p53-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5906)
TITLE p53-GFP
REFERENCE 2 (bases 1 to 5906)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5906
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 16..319
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 320..523
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 474..494
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 601..610
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 1801..1810
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1807..2526
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1852..1873)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2113..2132)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2460..2481
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2649..2770
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2686..2705)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2740..2759
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2777..3232)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2914..2935)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 3126..3145
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 3259..3363
/gene="bla"
/label=AmpR promoter
promoter 3365..3722
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3388..3408)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3573..3708
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3635..3654
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3757..4551
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3811..3830)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4421..4440
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4739..4758)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4783..4830
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5159..5747
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5648..5667
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
61 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
121 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
181 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
241 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
301 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
361 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
421 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
481 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
541 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacc
601 gccaccatgg aggagccgca gtcagatcct agcgtcgagc cccctctgag tcaggaaaca
661 ttttcagacc tatggaaact acttcctgaa aacaacgttc tgtccccctt gccgtcccaa
721 gcaatggatg atttgatgct gtccccggac gatattgaac aatggttcac tgaagaccca
781 ggtccagatg aagctcccag aatgccagag gctgctcccc gcgtggcccc tgcaccagca
841 gctcctacac cggcggcccc tgcaccagcc ccctcctggc ccctgtcatc ttctgtccct
901 tcccagaaaa cctaccaggg cagctacggt ttccgtctgg gcttcttgca ttctgggaca
961 gccaagtctg tgacttgcac gtactcccct gccctcaaca agatgttttg ccaactggcc
1021 aagacctgcc ctgtgcagct gtgggttgat tccacacccc cgcccggcac ccgcgtccgc
1081 gccatggcca tctacaagca gtcacagcac atgacggagg ttgtgaggcg ctgcccccac
1141 catgagcgct gctcagatag cgatggtctg gcccctcctc agcatcttat ccgagtggaa
1201 ggaaatttgc gtgtggagta tttggatgac agaaacactt ttcgacatag tgtggtggtg
1261 ccctatgagc cgcctgaggt tggctctgac tgtaccacca tccactacaa ctacatgtgt
1321 aacagttcct gcatgggcgg catgaaccgg aggcccatcc tcaccatcat cacactggaa
1381 gactccagtg gtaatctact gggacggaac agctttgagg tgcgtgtttg tgcctgtcct
1441 gggagagacc ggcgcacaga ggaagagaat ctccgcaaga aaggggagcc tcaccacgag
1501 ctgcccccag ggagcactaa gcgagcactg cccaacaaca ccagctcctc tccccagcca
1561 aagaagaaac cactggatgg agaatatttc acccttcaga tccgtgggcg tgagcgcttc
1621 gagatgttcc gagagctgaa tgaggccttg gaactcaagg atgcccaggc tgggaaggag
1681 ccagggggga gcagggctca ctccagccac ctgaagtcca aaaagggtca gtctacctcc
1741 cgccataaaa aactcatgtt caagacagaa gggcctgact cagacgggga tccaccggtc
1801 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
1861 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
1921 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
1981 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
2041 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
2101 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
2161 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
2221 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
2281 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
2341 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
2401 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
2461 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
2521 aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
2581 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
2641 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
2701 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
2761 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
2821 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
2881 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
2941 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
3001 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
3061 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
3121 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
3181 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
3241 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
3301 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
3361 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
3421 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
3481 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
3541 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
3601 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
3661 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
3721 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
3781 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
3841 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
3901 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
3961 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
4021 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
4081 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
4141 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
4201 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
4261 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
4321 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
4381 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
4441 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
4501 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
4561 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
4621 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
4681 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
4741 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
4801 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
4861 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
4921 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
4981 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
5041 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
5101 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
5161 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
5221 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
5281 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
5341 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
5401 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
5461 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
5521 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
5581 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
5641 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
5701 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
5761 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
5821 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca
5881 attacggggt cattagttca tagccc
//
