| 质粒名称: | pcDNA3 flag p53 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 10838 |
| 存储实验室: | Thomas Roberts |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | Invitrogen |
| 空载体大小: | 5500 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄青霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | p53 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1200 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | flag（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI（未销毁） |
| 3’克隆位点: | XbaI（未销毁） |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3 flag p53 was a gift from Thomas Roberts (Addgene plasmid # 10838 ; http://n2t.net/addgene:10838 ; RRID:Addgene_10838) |
| 参考文献部分: | Induction of p53-independent apoptosis by simian virus 40 small t antigen. Gjoerup O, Zaveri D, Roberts TM. J Virol. 2001 Oct . 75(19):9142-55. 10.1128/JVI.75.19.9142-9155.2001 PubMed 11533178 |
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 pcDNA3 flag p53 6590 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3 flag p53
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6590)
AUTHORS Gjoerup O, Zaveri D, Roberts TM
TITLE Induction of p53-independent apoptosis by simian virus 40 small t
antigen.
JOURNAL J Virol. 2001 Oct . 75(19):9142-55.
PUBMED 11533178
REFERENCE 2 (bases 1 to 6590)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6590
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 907..930
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 937..2115
/codon_start=1
/gene="Homo sapiens TP53"
/product="human tumor suppressor protein that is often
mutated in cancers"
/label=p53
/note="responds to cellular stresses to regulate the
expression of target genes"
/translation="EEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLS
PDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQG
SYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYK
QSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPP
EVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDR
RTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMF
RELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD"
promoter complement(2143..2161)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2144..2161)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2181..2198)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2187..2411
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2457..2885
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2544..2563)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2754..2775
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2894..2914)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2899..3228
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3079..3214
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3141..3160
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3295..4089
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3349..3368)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3959..3978
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4263..4384
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4300..4319)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4354..4373
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4433..4449)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4433..4449)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4446..4468)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4457..4473
/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(4481..4511)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4526..4547
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4664..4681)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4835..5423)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4915..4934)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5594..6454)
/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 6217..6236
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6455..6559)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
901 gacatggact acaaggacga cgatgacaag ggatccgagg agccgcagtc agatcctagc
961 gtcgagcccc ctctgagtca ggaaacattt tcagacctat ggaaactact tcctgaaaac
1021 aacgttctgt cccccttgcc gtcccaagca atggatgatt tgatgctgtc cccggacgat
1081 attgaacaat ggttcactga agacccaggt ccagatgaag ctcccagaat gccagaggct
1141 gctccccccg tggcccctgc accagcagct cctacaccgg cggcccctgc accagccccc
1201 tcctggcccc tgtcatcttc tgtcccttcc cagaaaacct accagggcag ctacggtttc
1261 cgtctgggct tcttgcattc tgggacagcc aagtctgtga cttgcacgta ctcccctgcc
1321 ctcaacaaga tgttttgcca actggccaag acctgccctg tgcagctgtg ggttgattcc
1381 acacccccgc ccggcacccg cgtccgcgcc atggccatct acaagcagtc acagcacatg
1441 acggaggttg tgaggcgctg cccccaccat gagcgctgct cagatagcga tggtctggcc
1501 cctcctcagc atcttatccg agtggaagga aatttgcgtg tggagtattt ggatgacaga
1561 aacacttttc gacatagtgt ggtggtgccc tatgagccgc ctgaggttgg ctctgactgt
1621 accaccatcc actacaacta catgtgtaac agttcctgca tgggcggcat gaaccggagg
1681 cccatcctca ccatcatcac actggaagac tccagtggta atctactggg acggaacagc
1741 tttgaggtgc gtgtttgtgc ctgtcctggg agagaccggc gcacagagga agagaatctc
1801 cgcaagaaag gggagcctca ccacgagctg cccccaggga gcactaagcg agcactgccc
1861 aacaacacca gctcctctcc ccagccaaag aagaaaccac tggatggaga atatttcacc
1921 cttcagatcc gtgggcgtga gcgcttcgag atgttccgag agctgaatga ggccttggaa
1981 ctcaaggatg cccaggctgg gaaggagcca ggggggagca gggctcactc cagccacctg
2041 aagtccaaaa agggtcagtc tacctcccgc cataaaaaac tcatgttcaa gacagaaggg
2101 cctgactcag actgactcga gcatgcatct agagggccct attctatagt gtcacctaaa
2161 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
2221 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
2281 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
2341 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
2401 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
2461 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
2521 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
2581 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
2641 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
2701 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
2761 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
2821 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
2881 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
2941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
3001 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
3061 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
3121 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
3181 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
3241 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
3301 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
3361 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
3421 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
3481 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
3541 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
3601 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
3661 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
3721 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
3781 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
3841 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
3901 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
3961 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
4021 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
4081 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
4141 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
4201 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
4261 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
4321 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
4381 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
4441 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
4501 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
4561 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
4621 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
4681 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
4741 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
4801 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
4861 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
4921 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
4981 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
5041 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
5101 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
5161 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
5221 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
5281 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
5341 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
5401 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
5461 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
5521 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
5581 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
5641 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
5701 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
5761 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
5821 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
5881 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
5941 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
6001 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
6061 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
6121 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
6181 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
6241 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
6301 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
6361 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
6421 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
6481 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
6541 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
