| 质粒名称: | Gal4-HA-HIF1a(530-652) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 24887 |
| 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 |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5252 bp |
| 完整质粒大小: | 6161 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: | hypoxia inducible factor1alpha |
| 物种来源: | 人源 |
| 插入大小: | 909 bp |
| 基因突变: | partial HIF1alpha cDNA (encoding aa 530-652)fused to Gal4 DNA binding domain, fused to HA-tag |
| 基因又名: |
Gal4-HA-HIF-ODD oxygen-dependent degradation domain |
| 标签/融合蛋白: |
Gal4dbd (N terminal on backbone)
HA-tag (N terminal on backbone)
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Hind III(未破坏) |
| 3' 克隆位点: | Xba I (未破坏) |
| 5' 测序引物 | CMV-forw or T7 |
| 3' 测序引物 | Sp6 |
|
文献引用方法 |
|
| 材料和方法部分: | Gal4-HA-HIF1α(530-652) was a gift from William Kaelin (爱迪基因 plasmid # 24887) |
| 参考文献部分: | HIFalpha targeted for VHL-mediated destruction by proline hydroxylation: implications for O2 sensing. Ivan M, Kondo K, Yang H, Kim W, Valiando J, Ohh M, Salic A, Asara JM, Lane WS, Kaelin WG. Science. 2001 Apr 20. 292(5516):464-8. 10.1126/science.1059817 PubMed 11292862 |
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 Gal4-HA-HIF1-alpha(530-652) 6261 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Gal4-HA-HIF1-alpha(530-652)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6261)
AUTHORS Ivan M, Kondo K, Yang H, Kim W, Valiando J, Ohh M, Salic A, Asara
JM, Lane WS, Kaelin WG
TITLE HIFalpha targeted for VHL-mediated destruction by proline
hydroxylation: implications for O2 sensing.
JOURNAL Science. 2001 Apr 20. 292(5516):464-8.
PUBMED 11292862
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|Gal4-HA-HIF1a(530-652)|
FEATURES Location/Qualifiers
source 1..6261
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 898..1338
/codon_start=1
/gene="Saccharomyces cerevisiae GAL4 (truncated)"
/product="DNA binding domain of the GAL4 transcriptional
activator"
/label=GAL4 DNA binding domain
/translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
CDS 1354..1380
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter complement(1813..1831)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(1814..1831)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(1851..1868)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1857..2081
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2127..2555
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2214..2233)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2424..2445
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2564..2584)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2569..2899
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2750..2885
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2812..2831
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2966..3760
/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(3020..3039)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3630..3649
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3934..4055
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3971..3990)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4025..4044
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4104..4120)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4104..4120)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4117..4139)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4128..4144
/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(4152..4182)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4197..4218
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4335..4352)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4506..5094)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4586..4605)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5265..6125)
/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 5888..5907
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6126..6230)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccatg
901 aagctactgt cttctatcga acaagcatgc gatatttgcc gacttaaaaa gctcaagtgc
961 tccaaagaaa aaccgaagtg cgccaagtgt ctgaagaaca actgggagtg tcgctactct
1021 cccaaaacca aaaggtctcc gctgactagg gcacatctga cagaagtgga atcaaggcta
1081 gaaagactgg aacagctatt tctactgatt tttcctcgag aagaccttga catgattttg
1141 aaaatggatt ctttacagga tataaaagca ttgttaacag gattatttgt acaagataat
1201 gtgaataaag atgccgtcac agatagattg gcttcagtgg agactgatat gcctctaaca
1261 ttgagacagc atagaataag tgcgacatca tcatcggaag agagtagtaa caaaggtcaa
1321 agacagttga ctgtatcgcc ggaattcatg gcttacccat acgatgttcc agattacgct
1381 agcttgggtg gtcatatggc catggaggcc ccggggatcc ccatggaatt caagttggaa
1441 ttggtagaaa aactttttgc tgaagacaca gaagcaaaga acccattttc tactcaggac
1501 acagatttag acttggagat gttagctccc tatatcccaa tggatgatga cttccagtta
1561 cgttccttcg atcagttgtc accattagaa agcagttccg caagccctga aagcgcaagt
1621 cctcaaagca cagttacagt attccagcag actcaaatac aagaacctac tgctaatgcc
1681 accactacca ctgccaccac tgatgaatta aaaacagtga caaaagaccg tatggaagac
1741 attaaaatat tgattgcatc tccatctcct acccacatac ataaagaaac tacttgatct
1801 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
1861 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
1921 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
1981 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
2041 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
2101 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
2161 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
2221 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
2281 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
2341 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
2401 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
2461 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
2521 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
2581 gggtgtggaa agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa
2641 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
2701 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
2761 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
2821 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
2881 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
2941 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
3001 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
3061 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
3121 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
3181 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
3241 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
3301 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
3361 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
3421 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
3481 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
3541 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
3601 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
3661 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
3721 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
3781 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
3841 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
3901 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
3961 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
4021 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
4081 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
4141 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
4201 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
4261 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
4321 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
4381 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
4441 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
4501 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
4561 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
4621 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
4681 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
4741 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
4801 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
4861 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
4921 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
4981 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
5041 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
5101 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
5161 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
5221 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
5281 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
5341 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
5401 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
5461 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
5521 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
5581 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
5641 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
5701 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
5761 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
5821 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
5881 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
5941 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
6001 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
6061 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
6121 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
6181 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
6241 cgaaaagtgc cacctgacgt c
//
