| 质粒名称: | pCMV Flag WT-PTEN |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 22231 |
| 存储实验室: | Hong Wu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV-Tag2 |
| 载体出品公司: | Stratagene |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4300 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: | PTEN |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1209 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: | BamH1(未破坏) |
| 3' 克隆位点: | EcoR1 (未破坏) |
| 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: | pCMV Flag WT-PTEN was a gift from Hong Wu (Addgene plasmid # 22231 ; http://n2t.net/addgene:22231 ; RRID:Addgene_22231) |
| 参考文献部分: | Analysis of PTEN complex assembly and identification of hnRNP C as a component of the PTEN-associated complex. Mosessian S, Avliyakulov NK, Mulholland DJ, Boontheung P, Loo JA, Wu H. J Biol Chem. 2009 Sep 9. ():. 10.1074/jbc.M109.027995 PubMed 19740742 |
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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 pCMV Flag WT-PTEN 5539 bp ds-DNA circular SYN 19-MAR-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCMV Flag WT-PTEN
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5539)
AUTHORS Mosessian S, Avliyakulov NK, Mulholland DJ, Boontheung P, Loo JA, Wu
H
TITLE Analysis of PTEN complex assembly and identification of hnRNP C as a
component of the PTEN-associated complex.
JOURNAL J Biol Chem. 2009 Sep 9. ():.
PUBMED 19740742
REFERENCE 2 (bases 1 to 5539)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 19, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5539
/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(1969..1985)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1970..1986)
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind complement(2041..2060)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(2042..2060)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 2334..2455
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2371..2390)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2425..2444
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2462..2917)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2599..2620)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2811..2830
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2944..3048
/gene="bla"
/label=AmpR promoter
promoter 3050..3407
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3073..3093)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3258..3393
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3320..3339
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3442..4236
/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(3496..3515)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4106..4125
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4424..4443)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4468..4515
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4844..5432
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5333..5352
/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 gataagagcc cgggcggatc
721 catgacagcc atcatcaaag agatcgttag cagaaacaaa aggagatatc aagaggatgg
781 attcgactta gacttgacct atatttatcc aaacattatt gctatgggat ttcctgcaga
841 aagacttgaa ggcgtataca ggaacaatat tgatgatgta gtaaggtttt tggattcaaa
901 gcataaaaac cattacaaga tatacaatct ttgtgctgaa agacattatg acaccgccaa
961 atttaattgc agagttgcac aatatccttt tgaagaccat aacccaccac agctagaact
1021 tatcaaaccc ttttgtgaag atcttgacca atggctaagt gaagatgaca atcatgttgc
1081 agcaattcac tgtaaagctg gaaagggacg aactggtgta atgatatgtg catatttatt
1141 acatcggggc aaatttttaa aggcacaaga ggccctagat ttctatgggg aagtaaggac
1201 cagagacaaa aagggagtaa ctattcccag tcagaggcgc tatgtgtatt attatagcta
1261 cctgttaaag aatcatctgg attatagacc agtggcactg ttgtttcaca agatgatgtt
1321 tgaaactatt ccaatgttca gtggcggaac ttgcaatcct cagtttgtgg tctgccagct
1381 aaaggtgaag atatattcct ccaattcagg acccacacga cgggaagaca agttcatgta
1441 ctttgagttc cctcagccgt tacctgtgtg tggtgatatc aaagtagagt tcttccacaa
1501 acagaacaag atgctaaaaa aggacaaaat gtttcacttt tgggtaaata cattcttcat
1561 accaggacca gaggaaacct cagaaaaagt agaaaatgga agtctatgtg atcaagaaat
1621 cgatagcatt tgcagtatag agcgtgcaga taatgacaag gaatatctag tacttacttt
1681 aacaaaaaat gatcttgaca aagcaaataa agacaaagcc aaccgatact tttctccaaa
1741 ttttaaggtg aagctgtact tcacaaaaac agtagaggag ccgtcaaatc cagaggctag
1801 cagttcaact tctgtaacac cagatgttag tgacaatgaa cctgatcatt atagatattc
1861 tgacaccact gactctgatc cagagaatga accttttgat gaagatcagc atacacaaat
1921 tacaaaagtc tgaaagcttg ggaagggcga attcgatatc aagcttatcg ataccgtcga
1981 cctcgagggg gggcccggta ccttaattaa ttaaggtacc aggtaagtgt acccaattcg
2041 ccctatagtg agtcgtatta caattcactc gatcgccctt cccaacagtt gcgcagcctg
2101 aatggcgaat ggagatccaa tttttaagtg tataatgtgt taaactactg attctaattg
2161 tttgtgtatt ttagattcac agtcccaagg ctcatttcag gcccctcagt cctcacagtc
2221 tgttcatgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
2281 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
2341 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
2401 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg
2461 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
2521 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
2581 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
2641 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
2701 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
2761 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
2821 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
2881 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
2941 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
3001 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagaatcc tgaggcggaa
3061 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
3121 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
3181 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
3241 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
3301 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
3361 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
3421 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
3481 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
3541 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
3601 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
3661 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
3721 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
3781 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
3841 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
3901 gatcaggatg atctggacga agaacatcag gggctcgcgc cagccgaact gttcgccagg
3961 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
4021 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
4081 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agaacttggc
4141 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
4201 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
4261 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
4321 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
4381 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
4441 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
4501 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
4561 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
4621 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
4681 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
4741 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
4801 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
4861 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
4921 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
4981 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
5041 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
5101 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
5161 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
5221 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
5281 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
5341 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
5401 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
5461 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
5521 tggataaccg tattaccgc
//
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