| 质粒名称: |
|
|---|---|
| 出品公司： | addgene |
| 目录编号: | 16639 |
| 存储实验室: | Bert Vogelstein |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV2-Tag 2A |
| 载体出品公司: | Stratagene |
| 空载体大小: | 4300 bp |
| 完整质粒大小: | 7516bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 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: | PIK3CA H1047R/Phosphatidylinositol 3-kinase /110 alpha |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 951 bp |
| 基因突变: | PIK3CA (a.k.a. CCM4, CLAPO, CLOVE, CWS5, MCAP, MCM, MCMTC, PI3K, PI3K-alpha, p110-alpha) |
| GenBank编号 | JN247409 |
| 启动子: | CMV |
| 标签/融合蛋白: | Flag (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | EcoRV / XmnI (destroyed during cloning) |
| 3’克隆位点: | XhoI (not destroyed) |
| 5’测序引物 | T3 |
| 3’测序引物 | T7 |
|
文献引用方法 |
|
| 材料和方法部分: | PIK3CA - Exon20 H1047R was a gift from Bert Vogelstein (Addgene plasmid # 16639 ; http://n2t.net/addgene:16639 ; RRID:Addgene_16639) |
| 参考文献部分: | High frequency of mutations of the PIK3CA gene in human cancers. Samuels Y, Wang Z, Bardelli A, Silliman N, Ptak J, Szabo S, Yan H, Gazdar A, Powell SM, Riggins GJ, Willson JK, Markowitz S, Kinzler KW, Vogelstein B, Velculescu VE. Science. 2004 Apr 23. 304(5670):554. 10.1126/science.1096502 PubMed 15016963 |
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 PIK3CA - Exon20 H1047R 7516 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS PIK3CA - Exon20 H1047R
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7516)
AUTHORS Samuels Y, Wang Z, Bardelli A, Silliman N, Ptak J, Szabo S, Yan H,
Gazdar A, Powell SM, Riggins GJ, Willson JK, Markowitz S, Kinzler
KW, Vogelstein B, Velculescu VE
TITLE High frequency of mutations of the PIK3CA gene in human cancers.
JOURNAL Science. 2004 Apr 23. 304(5670):554.
PUBMED 15016963
REFERENCE 2 (bases 1 to 7516)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7516
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 68..371
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 372..575
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 526..546
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
promoter 621..639
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
regulatory 674..683
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 683..706
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
primer_bind complement(4018..4037)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4019..4037)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 4311..4432
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4348..4367)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4402..4421
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(4439..4894)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4576..4597)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4788..4807
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4921..5025
/gene="bla"
/label=AmpR promoter
promoter 5027..5384
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(5050..5070)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 5235..5370
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5297..5316
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5419..6213
/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(5473..5492)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6083..6102
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(6401..6420)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 6445..6492
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6821..7409
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7310..7329
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
601 agcgattacg ccaagctcga aattaaccct cactaaaggg aacaaaagct ggagctccac
661 cgcggtggcg gccgccacca tggattacaa ggatgacgac gataaggccc gggcggatcc
721 cccgggctgc aggaattcga tagttcgaac catgcctcca agaccatcat caggtgaact
781 gtggggcatc cacttgatgc ccccaagaat cctagtagaa tgtttactac caaatggaat
841 gatagtgact ttagaatgcc tccgcgaggc tacattaata accataaagc atgaactatt
901 taaagaagca agaaaatacc ccctccatca acttcttcaa gatgaatctt cttacatttt
961 cgtaagtgtt actcaagaag cagaaaggga agaatttttt gatgaaacaa gacgactttg
1021 tgaccttcgg ctttttcaac cctttttaaa agtaattgaa ccagtaggca accgtgaaga
1081 aaagatcctc gatcgagaaa ttggttttgc tatcggcatg ccagtgtgtg aatttgatat
1141 ggttaaagat ccagaagtac aggacttccg aagaaatatt ctgaacgttt gtaaagaagc
1201 tgtggatctt agggacctca attcacctca tagtagagca atgtatgtct atcctccaaa
1261 tgtagaatct tcaccagaat cgccaaagca catatacaat aaattagata aagggcaaat
1321 aatagtggtg atctgggtaa tagtttctcc aaataatgac aagcagaagt atactctgaa
1381 aatcaaccat gactgtgtac cagaacaagt aattgctgaa gcaatcagga aaaaaactcg
1441 aagtatgttg ctatcctctg aacaactaaa actctgtgtt ttagaatatc agggcaagta
1501 tattttaaaa gtgtgtggat gtgatgaata cttcctagaa aaatatcctc tgagtcagta
1561 taagtatata agaagctgta taatgctagg gaggatgccc aatttgatgt tgatggctaa
1621 agaaagcctt tattctcaac tgccgatgga ctgtttcaca atgccatctt attccagacg
1681 catttccaca gctacaccat atatgaatgg agaagcatct acaaaatccc tttgggttat
1741 aaatagtgca ctcagaataa aaattctttg tgcaacctac gtgaatgtaa atattcgaga
1801 cattgataag atctatgttc gaacaggtat ctaccatgga ggagaaccct tatgtgacaa
1861 tgtgaacact caaagagtac cttgttccaa tcccaggtgg aatgaatggc tgaattatga
1921 tatatacatt cctgatcttc ctcgtgctgt tcgactttgc ccttccattt gctctgttaa
1981 aggccgaaag ggtgctaaag aggaacactg tccattggca tggggaaata taaacttgtt
2041 tgattacaca gacactctag tatctggaaa aatggctttg aatctttggc cagtacctca
2101 tggattagaa gatttgctga accctattgg tgttactgga tcaaatccaa ataaagaaac
2161 tccatgctta gagttggagt ttgactggtt cagcagtgtg gtaaagttcc cagatatgtc
2221 agtgattgaa gagcatgcca attggtctgt atcccgagaa gcaggattta gctattccca
2281 cgcaggactg agtaacagac tagctagaga caatgaatta agggaaaatg acaaagaaca
2341 gctcaaagca atttctacac gagatcctct ctctgaaatc actgagcagg agaaagattt
2401 tctatggagt cacagacact attgtgtaac tatccccgaa attctaccca aattgcttct
2461 gtctgttaaa tggaattcta gagatgaagt agcccagatg tattgcttgg taaaagattg
2521 gcctccaatc aaacctgaac aggctatgga acttctggac tgtaattacc cagatcctat
2581 ggttcgaggt tttgctgttc ggtgcttgga aaaatattta acagatgaca aactttctca
2641 gtatttaatt cagctagtac aggtcctaaa atatgaacaa tatttggata acttgcttgt
2701 gagattttta ctgaagaaag cattgactaa tcaaaggatt gggcactttt tcttttggca
2761 tttaaaatct gagatgcaca ataaaacagt tagccagagg tttggcctgc ttttggagtc
2821 ctattgtcgt gcatgtggga tgtatttgaa gcacctgaat aggcaagtcg aggcaatgga
2881 aaagctcatt aacttaactg acattctcaa acaggagaag aaggatgaaa cacaaaaggt
2941 acagatgaag tttttagttg agcaaatgag gcgaccagat ttcatggatg ctctacaggg
3001 ctttctgtct cctctaaacc ctgctcatca actaggaaac ctcaggcttg aagagtgtcg
3061 aattatgtcc tctgcaaaaa ggccactgtg gttgaattgg gagaacccag acatcatgtc
3121 agagttactg tttcagaaca atgagatcat ctttaaaaat ggggatgatt tacggcaaga
3181 tatgctaaca cttcaaatta ttcgtattat ggaaaatatc tggcaaaatc aaggtcttga
3241 tcttcgaatg ttaccttatg gttgtctgtc aatcggtgac tgtgtgggac ttattgaggt
3301 ggtgcgaaat tctcacacta ttatgcaaat tcagtgcaaa ggcggcttga aaggtgcact
3361 gcagttcaac agccacacac tacatcagtg gctcaaagac aagaacaaag gagaaatata
3421 tgatgcagcc attgacctgt ttacacgttc atgtgctgga tactgtgtag ctaccttcat
3481 tttgggaatt ggagatcgtc acaatagtaa catcatggtg aaagacgatg gacaactgtt
3541 tcatatagat tttggacact ttttggatca caagaagaaa aaatttggtt ataaacgaga
3601 acgtgtgcca tttgttttga cacaggattt cttaatagtg attagtaaag gagcccaaga
3661 atgcacaaag acaagagaat ttgagaggtt tcaggagatg tgttacaagg cttatctagc
3721 tattcgacag catgccaatc tcttcataaa tcttttctca atgatgcttg gctctggaat
3781 gccagaacta caatcttttg atgacattgc atacattcga aagaccctag ccttagataa
3841 aactgagcaa gaggctttgg agtatttcat gaaacaaatg aatgatgcac gtcatggtgg
3901 ctggacaaca aaaatggatt ggatcttcca cacaattaaa cagcatgcat tgaactagct
3961 cgaggggggg cccggtacct taattaatta aggtaccagg taagtgtacc caattcgccc
4021 tatagtgagt cgtattacaa ttcactcgat cgcccttccc aacagttgcg cagcctgaat
4081 ggcgaatgga gatccaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt
4141 gtgtatttta gattcacagt cccaaggctc atttcaggcc cctcagtcct cacagtctgt
4201 tcatgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
4261 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
4321 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
4381 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
4441 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
4501 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
4561 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
4621 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
4681 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
4741 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
4801 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
4861 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
4921 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
4981 caataaccct gataaatgct tcaataatat tgaaaaagga agaatcctga ggcggaaaga
5041 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
5101 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
5161 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
5221 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
5281 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
5341 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
5401 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
5461 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
5521 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
5581 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
5641 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
5701 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
5761 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
5821 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
5881 caggatgatc tggacgaaga acatcagggg ctcgcgccag ccgaactgtt cgccaggctc
5941 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
6001 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
6061 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga acttggcggc
6121 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
6181 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
6241 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
6301 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
6361 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
6421 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
6481 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
6541 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
6601 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
6661 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
6721 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
6781 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
6841 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
6901 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
6961 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
7021 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
7081 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
7141 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
7201 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
7261 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
7321 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
7381 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
7441 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
7501 ataaccgtat taccgc
//
