| 质粒名称: | HA-HIF1alpha-pcDNA3 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 18949 |
| 存储实验室: | William Kaelin |
|
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: | 5433 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） |
| 载体抗性: | 氨苄霉素100 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: | Hypoxia inducible factor 1 alpha,HIF1a |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2505 bp |
| 基因突变: | wt cDNA |
| GenBank编号 | NM_001530 |
| 启动子: | -- |
| 标签/融合蛋白: | HA标签（主干上的N终端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamH I(未破坏) |
| 3’克隆位点: | EcoR I（克隆期间销毁） |
| 5’测序引物 | T7 |
| 3’测序引物 | SP6 |
|
文献引用方法 |
|
| 材料和方法部分: | HA-HIF1alpha-pcDNA3 was a gift from William Kaelin (Addgene plasmid # 18949 ; http://n2t.net/addgene:18949 ; RRID:Addgene_18949) |
| 参考文献部分: | Inhibition of HIF is necessary for tumor suppression by the von Hippel-Lindau protein. Kondo K, Klco J, Nakamura E, Lechpammer M, Kaelin WG. Cancer Cell. 2002 Apr . 1(3):237-46. 10.1016/S1535-6108(02)00043-0 PubMed 12086860 |
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 HA-HIF1alpha-pcDNA3 7937 bp ds-DNA circular SYN 16-AUG-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS HA-HIF1alpha-pcDNA3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7937)
AUTHORS Kondo K, Klco J, Nakamura E, Lechpammer M, Kaelin WG
TITLE Inhibition of HIF is necessary for tumor suppression by the von
Hippel-Lindau protein.
JOURNAL Cancer Cell. 2002 Apr . 1(3):237-46.
PUBMED 12086860
REFERENCE 2 (bases 1 to 7937)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 16, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7937
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(46..65)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 237..616
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 617..820
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 771..791
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 865..884
/label=T7
/note="T7 promoter, forward primer"
promoter 865..883
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 906..932
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
regulatory 945..954
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter complement(3492..3510)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3493..3510)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(3530..3547)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3536..3760
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3806..4234
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3893..3912)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4103..4124
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4243..4263)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4248..4577
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4428..4563
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4490..4509
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4644..5438
/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(4698..4717)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5308..5327
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5612..5733
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5649..5668)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5703..5722
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5782..5798)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5782..5798)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5795..5817)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5806..5822
/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(5830..5860)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5875..5896
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6013..6030)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6184..6772)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6264..6283)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6943..7803)
/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 7566..7585
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7804..7908)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttacca
901 tggcctaccc ctacgacgtg cccgactacg cctccctcgg atccgccacc atggagggcg
961 ccggcggcgc gaacgacaag aaaaagataa gttctgaacg tcgaaaagaa aagtctcgag
1021 atgcagccag atctcggcga agtaaagaat ctgaagtttt ttatgagctt gctcatcagt
1081 tgccacttcc acataatgtg agttcgcatc ttgataaggc ctctgtgatg aggcttacca
1141 tcagctattt gcgtgtgagg aaacttctgg atgctggtga tttggatatt gaagatgaca
1201 tgaaagcaca gatgaattgc ttttatttga aagccttgga tggttttgtt atggttctca
1261 cagatgatgg tgacatgatt tacatttctg ataatgtgaa caaatacatg ggattaactc
1321 agtttgaact aactggacac agtgtgtttg attttactca tccatgtgac catgaggaaa
1381 tgagagaaat gcttacacac agaaatggcc ttgtgaaaaa gggtaaagaa caaaacacac
1441 agcgaagctt ttttctcaga atgaagtgta ccctaactag ccgaggaaga actatgaaca
1501 taaagtctgc aacatggaag gtattgcact gcacaggcca cattcacgta tatgatacca
1561 acagtaacca acctcagtgt gggtataaga aaccacctat gacctgcttg gtgctgattt
1621 gtgaacccat tcctcaccca tcaaatattg aaattccttt agatagcaag actttcctca
1681 gtcgacacag cctggatatg aaattttctt attgtgatga aagaattacc gaattgatgg
1741 gatatgagcc agaagaactt ttaggccgct caatttatga atattatcat gctttggact
1801 ctgatcatct gaccaaaact catcatgata tgtttactaa aggacaagtc accacaggac
1861 agtacaggat gcttgccaaa agaggtggat atgtctgggt tgaaactcaa gcaactgtca
1921 tatataacac caagaattct caaccacagt gcattgtatg tgtgaattac gttgtgagtg
1981 gtattattca gcacgacttg attttctccc ttcaacaaac agaatgtgtc cttaaaccgg
2041 ttgaatcttc agatatgaaa atgactcagc tattcaccaa agttgaatca gaagatacaa
2101 gtagcctctt tgacaaactt aagaaggaac ctgatgcttt aactttgctg gccccagccg
2161 ctggagacac aatcatatct ttagattttg gcagcaacga cacagaaact gatgaccagc
2221 aacttgagga agtaccatta tataatgatg taatgctccc ctcacccaac gaaaaattac
2281 agaatataaa tttggcaatg tctccattac ccaccgctga aacgccaaag ccacttcgaa
2341 gtagtgctga ccctgcactc aatcaagaag ttgcattaaa attagaacca aatccagagt
2401 cactggaact ttcttttacc atgccccaga ttcaggatca gacacctagt ccttccgatg
2461 gaagcactag acaaagttca cctgagccta atagtcccag tgaatattgt ttttatgtgg
2521 atagtgatat ggtcaatgaa ttcaagttgg aattggtaga aaaacttttt gctgaagaca
2581 cagaagcaaa gaacccattt tctactcagg acacagattt agacttggag atgttagctc
2641 cctatatccc aatggatgat gacttccagt tacgttcctt cgatcagttg tcaccattag
2701 aaagcagttc cgcaagccct gaaagcgcaa gtcctcaaag cacagttaca gtattccagc
2761 agactcaaat acaagaacct actgctaatg ccaccactac cactgccacc actgatgaat
2821 taaaaacagt gacaaaagac cgtatggaag acattaaaat attgattgca tctccatctc
2881 ctacccacat acataaagaa actactagtg ccacatcatc accatataga gatactcaaa
2941 gtcggacagc ctcaccaaac agagcaggaa aaggagtcat agaacagaca gaaaaatctc
3001 atccaagaag ccctaacgtg ttatctgtcg ctttgagtca aagaactaca gttcctgagg
3061 aagaactaaa tccaaagata ctagctttgc agaatgctca gagaaagcga aaaatggaac
3121 atgatggttc actttttcaa gcagtaggaa ttggaacatt attacagcag ccagacgatc
3181 atgcagctac tacatcactt tcttggaaac gtgtaaaagg atgcaaatct agtgaacaga
3241 atggaatgga gcaaaagaca attattttaa taccctctga tttagcatgt agactgctgg
3301 ggcaatcaat ggatgaaagt ggattaccac agctgaccag ttatgattgt gaagttaatg
3361 ctcctataca aggcagcaga aacctactgc agggtgaaga attactcaga gctttggatc
3421 aagttaactg acaattctgc agatatccat cacactggcg gccgctcgag catgcatcta
3481 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
3541 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
3601 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
3661 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
3721 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
3781 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
3841 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
3901 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
3961 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
4021 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
4081 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
4141 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
4201 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
4261 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
4321 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
4381 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
4441 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
4501 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
4561 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
4621 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
4681 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
4741 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
4801 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
4861 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
4921 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
4981 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
5041 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
5101 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
5161 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
5221 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
5281 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
5341 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
5401 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
5461 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
5521 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
5581 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
5641 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
5701 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
5761 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
5821 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
5881 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
5941 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
6001 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
6061 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
6121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
6181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
6241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
6301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
6361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
6421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
6481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
6541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
6601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
6661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
6721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
6781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
6841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
6901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
6961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
7021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
7081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
7141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
7201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
7261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
7321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
7381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
7441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
7501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
7561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
7621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
7681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
7741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
7801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
7861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
7921 aaaagtgcca cctgacg
//
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