| 质粒名称: | pLNCX2 ER:ras |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 67844 |
| 存储实验室: | Masashi Narita |
|
载体骨架基本信息 |
|
| 载体名称: | pLNCX2 |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6100 bp |
| 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: | ER1a H-RasG12V fusion |
| 物种来源: | H. sapiens (human), M. musculus (mouse) |
| 插入大小: | 1600 bp |
| 基因突变: | G525R renders the HBD largely insensitive to 17ß‐estradiol at concentrations less than 100 nM but does not abrogate its responsiveness to the synthetic ligand 4‐hydroxytamoxifen |
| 启动子 | CMV |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | HindIII(未破坏) |
| 3' 克隆位点: | HindIII(未破坏) |
| 5' 测序引物 | AGCTCGTTTAGTGAACCGTCAGATC |
| 3' 测序引物 | ACCTACAGGTGGGGTCTTTCATTCCC |
|
文献引用方法 |
|
| 材料和方法部分: | pLNCX2 ER:ras was a gift from Masashi Narita (Addgene plasmid # 67844 ; http://n2t.net/addgene:67844 ; RRID:Addgene_67844) |
| 参考文献部分: | Autophagy mediates the mitotic senescence transition. Young AR, Narita M, Ferreira M, Kirschner K, Sadaie M, Darot JF, Tavare S, Arakawa S, Shimizu S, Watt FM, Narita M. Genes Dev. 2009 Apr 1;23(7):798-803. doi: 10.1101/gad.519709. Epub 2009 Mar 11. 10.1101/gad.519709 PubMed 19279323 |
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 pLNCX2 ER:ras 7715 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION 4-hydroxytamoxifen inducible ER:ras fusion protein.
ACCESSION .
VERSION .
KEYWORDS pLNCX2 ER:ras
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7715)
AUTHORS Young AR, Narita M, Ferreira M, Kirschner K, Sadaie M, Darot JF,
Tavare S, Arakawa S, Shimizu S, Watt FM, Narita M
TITLE Autophagy mediates the mitotic senescence transition.
JOURNAL Genes Dev. 2009 Apr 1;23(7):798-803. doi: 10.1101/gad.519709. Epub
2009 Mar 11.
PUBMED 19279323
REFERENCE 2 (bases 1 to 7715)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7715
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 414..589
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 652..851
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1052..1468
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1398..1420
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1436..1452
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
CDS 1512..2306
/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(1566..1585)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 2176..2195
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
enhancer 2378..2681
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2682..2885
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2836..2856
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 3937..4506
/codon_start=1
/product="human oncoprotein generated by the G12V mutation
in the small GTPase H-Ras"
/label=H-Ras (G12V)
/translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
RQHKLRKLNPPDESGPGCMSCKCVLS"
LTR 4617..5210
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(5313..5335)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 5421..5561
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 5476..5495
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(5576..5593)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5747..6335)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5827..5846)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6506..7366)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 7129..7148
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7367..7471)
/gene="bla"
/label=AmpR promoter
primer_bind 7539..7557
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tttgaaagac cccacccgta ggtggcaagc tagcttaagt aacgccactt tgcaaggcat
61 ggaaaaatac ataactgaga atagaaaagt tcagatcaag gtcaggaaca aagaaacagc
121 tgaataccaa acaggatatc tgtggtaagc ggttcctgcc ccggctcagg gccaagaaca
181 gatgagacag ctgagtgatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
241 ctcggggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagtgaa
301 tcatcagatg tttccagggt gccccaagga cctgaaaatg accctgtacc ttatttgaac
361 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc cgctctccga gctcaataaa
421 agagcccaca acccctcact cggcgcgcca gtcttccgat agactgcgtc gcccgggtac
481 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggtctcg ctgttccttg
541 ggagggtctc ctctgagtga ttgactaccc acgacggggg tctttcattt gggggctcgt
601 ccgggatttg gagacccctg cccagggacc accgacccac caccgggagg taagctggcc
661 agcaacttat ctgtgtctgt ccgattgtct agtgtctatg tttgatgtta tgcgcctgcg
721 tctgtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt
781 ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg
841 cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt
901 aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa
961 ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct
1021 gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt
1081 gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa
1141 gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc
1201 gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc
1261 tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt
1321 tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc
1381 atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta
1441 tccagccctc actccttctc taggcgccgg aattccgatc tgatcaagag acaggatgag
1501 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
1561 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
1621 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
1681 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
1741 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
1801 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
1861 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
1921 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
1981 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
2041 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
2101 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
2161 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
2221 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
2281 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgtaatag taatcaatta
2341 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
2401 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
2461 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
2521 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
2581 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
2641 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
2701 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
2761 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
2821 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
2881 gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagctcaa
2941 gcttgttgat ccaccatgcg aaatgaaatg ggtgcttcag gagacatgag ggctgccaac
3001 ctttggccaa gccctcttgt gattaagcac actaagaaga atagccctgc cttgtccttg
3061 acagctgacc agatggtcag tgccttgttg gatgctgaac cgcccatgat ctattctgaa
3121 tatgatcctt ctagaccctt cagtgaagcc tcaatgatgg gcttattgac caacctagca
3181 gatagggagc tggttcatat gatcaactgg gcaaagagag tgccaggctt tggggacttg
3241 aatctccatg atcaggtcca ccttctcgag tgtgcctggc tggagattct gatgattggt
3301 ctcgtctggc gctccatgga acacccgggg aagctcctgt ttgctcctaa cttgctcctg
3361 gacaggaatc aaggtaaatg tgtggaaggc atggtggaga tctttgacat gttgcttgct
3421 acgtcaagtc ggttccgcat gatgaacctg cagggtgaag agtttgtgtg cctcaaatcc
3481 atcattttgc ttaattccgg agtgtacacg tttctgtcca gcaccttgaa gtctctggaa
3541 gagaaggacc acatccaccg tgtcctggac aagatcacag acactttgat ccacctgatg
3601 gccaaagctg gcctgactct gcagcagcag catcgccgcc tagctcagct ccttctcatt
3661 ctttcccata tccggcatat gagtaacaaa cgcatggagc atctctacaa catgaaatgc
3721 aagaacgtgg tacccctcta tgacctgctc ctggagatgt tggatgccca ccgccttcat
3781 gccccagcca gtcgcatggg agtgccccca gaggagccca gccagaccca gctggccacc
3841 accagctcca cttcagcaca ttccttacaa acctactaca tacccccgga agcagagggc
3901 ttccccaacg atatcccacg tggaggcctg aattccatga cggaatataa gctggtggtg
3961 gtgggcgccg tcggtgtggg caagagtgcg ctgaccatcc agctgatcca gaaccatttt
4021 gtggacgaat acgaccccac tatagaggat tcctaccgga agcaggtggt cattgatggg
4081 gagacgtgcc tgttggacat cctggatacc gccggccagg aggagtacag cgccatgcgg
4141 gaccagtaca tgcgcaccgg ggagggcttc ctgtgtgtgt ttgccatcaa caacaccaag
4201 tcttttgagg acatccacca gtacagggag cagatcaaac gggtgaagga ctcggatgac
4261 gtgcccatgg tgctggtggg gaacaagtgt gacctggctg cacgcactgt ggaatctcgg
4321 caggctcagg acctcgcccg aagctacggc atcccctaca tcgagacctc ggccaagacc
4381 cggcagggag tggaggatgc cttctacacg ttggtgcgtg agatccggca gcacaagctg
4441 cggaagctga accctcctga tgagagtggc cccggctgca tgagctgcaa gtgtgtgctc
4501 tcctgacgca attcgatatc aagcttgttt ggccgaggcg gccgcttgtc gacaggcctt
4561 aatggcctaa catcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
4621 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
4681 aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct
4741 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
4801 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
4861 ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag
4921 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
4981 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
5041 aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta
5101 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
5161 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg
5221 tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc
5281 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
5341 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
5401 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
5461 actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
5521 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
5581 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
5641 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
5701 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
5761 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
5821 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
5881 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
5941 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
6001 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
6061 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
6121 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
6181 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
6241 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
6301 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
6361 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
6421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
6481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
6541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
6601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
6661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
6721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
6781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
6841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
6901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
6961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
7021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
7081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
7141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
7201 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
7261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
7321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
7381 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
7441 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
7501 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
7561 ttcgtcttca agaattcata ccagatcacc gaaaactgtc ctccaaatgt gtccccctca
7621 cactcccaaa ttcgcgggct tctgcctctt agaccactct accctattcc ccacactcac
7681 cggagccaaa gccgcggccc ttccgtttct ttgct
//
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