| 质粒名称: | pcDNA3-H-Ras_V12 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 39504 |
| 存储实验室: | Julian Downward |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 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: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | HRAS V12 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | G12V |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI(未破坏) |
| 3' 克隆位点: | EcoRI(未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3-H-Ras_V12 was a gift from Julian Downward (Addgene plasmid # 39504 ; http://n2t.net/addgene:39504 ; RRID:Addgene_39504) |
| 参考文献部分: | Role of phosphoinositide 3-OH kinase in cell transformation and control of the actin cytoskeleton by Ras. Rodriguez-Viciana P, Warne PH, Khwaja A, Marte BM, Pappin D, Das P, Waterfield MD, Ridley A, Downward J. Cell. 1997 May 2;89(3):457-67. 10.1016/S0092-8674(00)80226-3 PubMed 9150145 |
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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 pcDNA3-H-Ras_V12 6340 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3-H-Ras_V12
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6340)
AUTHORS Rodriguez-Viciana P, Warne PH, Khwaja A, Marte BM, Pappin D, Das P,
Waterfield MD, Ridley A, Downward J
TITLE Role of phosphoinositide 3-OH kinase in cell transformation and
control of the actin cytoskeleton by Ras.
JOURNAL Cell. 1997 May 2;89(3):457-67.
PUBMED 9150145
REFERENCE 2 (bases 1 to 6340)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6340
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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)."
primer_bind 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 270..391
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(281..300)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 335..354
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(565..1359)
/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(676..695)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 1286..1305
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(1426..1755)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 1440..1575
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(1494..1513)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 1740..1760
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(1769..2197)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1879..1900)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2091..2110
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 2243..2467
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 2456..2473
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
promoter 2493..2511
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2493..2510
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(3522..3541)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3523..3541)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(3586..3789)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(3615..3635)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 3790..4169
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 4341..4360
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 4435..4539
/gene="bla"
/label=AmpR promoter
CDS 4540..5400
/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 complement(4758..4777)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5571..6159
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6060..6079
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6313..6330
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
481 catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc gcttggtcgg
541 tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat
601 gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc
661 gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac
721 acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg
781 caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag
841 cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc
901 gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc
961 gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga
1021 tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa
1081 tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc
1141 cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga
1201 caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc
1261 atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc
1321 ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt
1381 ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc aaaagcctag
1441 gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg cggcctcggc
1501 ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg aactgggcgg
1561 agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact aattgagatg
1621 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc tggttgctga
1681 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
1741 ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta aatcagctca
1801 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
1861 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
1921 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
1981 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
2041 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
2101 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
2161 acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct agagccccag
2221 ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg cctgctattg
2281 tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat agaatgacac
2341 ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt gggagtggca
2401 ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg gcaactagaa
2461 ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag aatagggccc
2521 tctagatgca tgctcgagcg gccgccagtg tgatggatat ctgcagaatt cccgatctcg
2581 accttttttt ttttttttca agaccatcca ataatttact gtgatcccat ctgtgcccga
2641 caagggccca cagaggcctg ggaggggagc taagggctgg ggttccggtg gcatttggga
2701 tgttcaagac agtctgtgca cagcctccct gggagggtct gcagtcacct cggcccacgg
2761 tcccggggtg actgggctcc agcagccctt ccttccttcc ttgcttccgt ccttccttcc
2821 tcctccttcc gtctgcacct ccttcctgca tccggcacct ccatgtcctg agcttgtgct
2881 gcgtcaggag agcacacact tgcagctcat gcagccgggg ccactctcat caggagggtt
2941 cagcttccgc agcttgtgct gccggatctc acgcaccaac gtgtagaagg catcctccac
3001 tccctgccgg gtcttggccg aggtctcgat gtaggggatg ccgtagcttc gggcgaggtc
3061 ctgagcctgc cgagattcca cagtgcgtgc agccaggtca cacttgttcc ccaccagcac
3121 catgggcacg tcatccgagt ccttcacccg tttgatctgc tccctgtact ggtggatgtc
3181 ctcaaaagac ttggtgttgt tgatggcaaa cacacacagg aagccctccc cggtgcgcat
3241 gtactggtcc cgcatggcgc tgtactcctc ctggccggcg gtatccagga tgtccaacag
3301 gcacgtctcc ccatcaatga ccacctgctt ccggtaggaa tcctctatag tggggtcgta
3361 ttcgtccaca aaatggttct ggatcagctg gatggtcagc gcactcttgc ccacaccgac
3421 ggcgccaaca acaacaagct tgtattcggt catagatcgg gaattccagc acactggcgg
3481 ccgttactag tggatccgag ctcggtacca agcttgggtc tccctatagt gagtcgtatt
3541 aatttcgata agccagtaag cagtgggttc tctagttagc cagagagctc tgcttatata
3601 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
3661 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
3721 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
3781 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
3841 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
3901 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
3961 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
4021 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
4081 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
4141 ctattaataa ctagtcaata atcaatgtca acgcgtatat ctggcccgta catcgcgaag
4201 cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa ttgtcggtca agccttgcct
4261 tgttgtagct taaattttgc tcgcgcacta ctcagcgacc tccaacacac aagcagggag
4321 cagatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatagggg
4381 atcgggagat ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga
4441 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
4501 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
4561 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
4621 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
4681 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
4741 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
4801 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
4861 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
4921 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
4981 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
5041 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
5101 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
5161 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
5221 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
5281 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
5341 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
5401 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
5461 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
5521 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
5581 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
5641 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
5701 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
5761 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
5821 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
5881 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
5941 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
6001 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
6061 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
6121 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
6181 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
6241 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
6301 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
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