| 质粒名称: | pcDNA3 mHIF-1α MYC (P402A/P577A/N813A) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 44028 |
| 存储实验室: | Celeste Simon |
|
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: | 5446 bp |
| 完整质粒大小: | 7950 bp |
| 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 |
| 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: | mHIF-1α (P402A/P577A/N813A),HIF-1α,Hypoxia inducible factor 1, alpha subunit,HIF1alpha |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 2500 bp |
| 基因突变: | P402A, P577A, N813A |
| GenBank编号 | NM_010431.2 NP_034561.2 |
| 启动子: | CMV |
| 标签/融合蛋白: | Myc（插入件上的C端子） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI(未破坏) |
| 3’克隆位点: | -- |
| 5’测序引物 | CMV-F; T7 |
| 3’测序引物 | BGH-rev; Sp6 |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3 mHIF-1α MYC (P402A/P577A/N813A) was a gift from Celeste Simon (Addgene plasmid # 44028 ; http://n2t.net/addgene:44028 ; RRID:Addgene_44028) |
| 参考文献部分: | The N-terminal transactivation domain confers target gene specificity of hypoxia-inducible factors HIF-1alpha and HIF-2alpha. Hu CJ, Sataur A, Wang L, Chen H, Simon MC. Mol Biol Cell. 2007 Nov;18(11):4528-42. Epub 2007 Sep 5. 10.1091/mbc.e06-05-0419 PubMed 17804822 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS pcDNA3 mHIF-1-alpha MYC (P402A/P577A/N813A) 7894 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA3 mHIF-1-alpha MYC (P402A/P577A/N813A)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7894)
AUTHORS Hu CJ, Sataur A, Wang L, Chen H, Simon MC
TITLE The N-terminal transactivation domain confers target gene
specificity of hypoxia-inducible factors HIF-1alpha and HIF-2alpha.
JOURNAL Mol Biol Cell. 2007 Nov;18(11):4528-42. Epub 2007 Sep 5.
PUBMED 17804822
REFERENCE 2 (bases 1 to 7894)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7894
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(11..28)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(182..770)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(262..281)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(941..1801)
/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 1564..1583
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1802..1906)
/gene="bla"
/label=AmpR promoter
primer_bind complement(1981..2000)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 2172..2551
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2552..2755
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2706..2726
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2800..2819
/label=T7
/note="T7 promoter, forward primer"
promoter 2800..2818
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS complement(5284..5295)
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
CDS 5358..5387
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(5422..5439)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5428..5652
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 5698..6126
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5785..5804)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5995..6016
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6135..6155)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6140..6469
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6320..6455
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6382..6401
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 6536..7330
/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
VLFVKTDLSGALNGLQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(6590..6609)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 7200..7219
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 7504..7625
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7541..7560)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 7595..7614
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(7674..7690)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7674..7690)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7687..7709)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7698..7714
/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(7722..7752)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7767..7788
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
361 gcgtagcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
2821 gacccaagct tggtaccgag ctcggatcca tggagggcgc cggcggcgag aacgagaaga
2881 aaaagatgag ttctgaacgt cgaaaagaaa agtctagaga tgcagcaaga tctcggcgaa
2941 gcaaagagtc tgaagttttt tatgagcttg ctcatcagtt gccacttccc cacaatgtga
3001 gctcacatct tgataaagct tctgttatga ggctcaccat cagttattta cgtgtgagaa
3061 aacttctgga tgccggtggt ctagacagtg aagatgagat gaaggcacag atggactgtt
3121 tttatctgaa agccctagat ggctttgtga tggtgctaac agatgacggc gacatggttt
3181 acatttctga taacgtgaac aaatacatgg ggttaactca gtttgaacta actggacaca
3241 gtgtgtttga ttttactcat ccatgtgacc atgaggaaat gagagaaatg cttacacaca
3301 gaaatggccc agtgagaaaa gggaaagaac taaacacaca gcggagcttt tttctcagaa
3361 tgaagtgcac cctaacaagc cgggggagga cgatgaacat caagtcagca acgtggaagg
3421 tgcttcactg cacgggccat attcatgtct atgataccaa cagtaaccaa cctcagtgtg
3481 ggtacaagaa accacccatg acgtgcttgg tgctgatttg tgaacccatt cctcatccgt
3541 caaatattga aattccttta gatagcaaga catttctcag tcgacacagc ctcgatatga
3601 aattttctta ctgtgatgaa agaattactg agttgatggg ttatgagccg gaagaacttt
3661 tgggccgctc aatttatgaa tattatcatg ctttggattc tgatcatctg accaaaactc
3721 accatgatat gtttactaaa ggacaagtca ccacaggaca gtacaggatg cttgccaaaa
3781 gaggtggata tgtctgggtt gaaactcaag caactgtcat atataatacg aagaactccc
3841 agccacagtg cattgtgtgt gtgaattatg ttgtaagtgg tattattcag cacgacttga
3901 ttttctccct tcaacaaaca gaatctgtgc tcaaaccagt tgaatcttca gatatgaaga
3961 tgactcagct gttcaccaaa gttgaatcag aggatacaag ctgccttttt gataagctta
4021 agaaggagcc tgatgctctc actctgctgg ctgccgctgc cggcgacacc atcatctctc
4081 tggattttgg cagcgatgac acagaaactg aagatcaaca acttgaagat gttccattat
4141 ataatgatgt aatgtttccc tcttctaatg aaaaattaaa tataaacctg gcaatgtctc
4201 ctttaccttc atcggaaact ccaaagccac ttcgaagtag tgctgatcct gcactgaatc
4261 aagaggttgc attaaaatta gaatcaagtc cagagtcact gggactttct tttaccatgc
4321 cccagattca agatcagcca gcaagtcctt ctgatggaag cactagacaa agttcacctg
4381 agagacttct tcaggaaaac gtaaacactc ctaacttttc ccagcctaac agtcccagtg
4441 aatattgctt tgatgtggat agcgatatgg tcaatgtatt taagttggaa ctggtggaaa
4501 aactgtttgc tgaagacaca gaggcaaaga atccattttc aactcaggac actgatttag
4561 atttggagat gctggctgcc tatatcccaa tggatgatga tttccagtta cgttcctttg
4621 atcagttgtc accattagag agcaattctc caagccctcc aagtatgagc acagttactg
4681 ggttccagca gacccagtta cagaaaccta ccatcactgc cactgccacc acaactgcca
4741 ccactgatga atcaaaaaca gagacgaagg acaataaaga agatattaaa atactgattg
4801 catctccatc ttctacccaa gtacctcaag aaacgaccac tgctaaggca tcagcataca
4861 gtggcactca cagtcggaca gcctcaccag acagagcagg aaagagagtc atagaacaga
4921 cagacaaagc tcatccaagg agccttaacc tgtctgccac tttgaatcaa agaaatactg
4981 ttcctgagga agaattaaac ccaaagacaa tagcttcgca gaatgctcag aggaagcgaa
5041 aaatggaaca tgatggctcc ctttttcaag cagcaggaat tggaacatta ttgcagcaac
5101 caggtgactg tgcacctact atgtcacttt cctggaaacg agtgaaagga ttcatatcta
5161 gtgaacagaa tggaacggag caaaagacta ttattttaat accctccgat ttagcatgca
5221 gactgctggg gcagtcaatg gatgagagtg gattaccaca gctgaccagt tacgattgtg
5281 aagttgcagc tcccatacaa ggcagcagaa acctactgca gggtgaagaa ttactcagag
5341 ctttggatca agttaacgaa caaaaactca tctcagaaga ggatctgtga ggatccgggc
5401 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
5461 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
5521 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
5581 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
5641 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
5701 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
5761 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
5821 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
5881 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
5941 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
6001 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
6061 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
6121 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
6181 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
6241 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
6301 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
6361 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
6421 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
6481 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
6541 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
6601 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
6661 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg gactgcagga
6721 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
6781 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
6841 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
6901 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
6961 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
7021 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
7081 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
7141 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
7201 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
7261 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
7321 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
7381 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
7441 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
7501 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
7561 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
7621 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
7681 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
7741 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
7801 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
7861 cgcgcgggga gaggcggttt gcgtattggg cgct
//
