| 质粒名称: | Flag-Keap1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 28023 |
| 存储实验室: | Qing Zhong |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA/FRT/TO |
| 载体出品公司: | Invitrogen |
| 空载体大小: | 5137 bp |
| 完整质粒大小: | 7070bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | Hygromycin |
| 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: | Keap1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1847 bp |
| 标签/融合蛋白: | 3xFlag (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | Flag-Keap1 was a gift from Qing Zhong (Addgene plasmid # 28023 ; http://n2t.net/addgene:28023 ; RRID:Addgene_28023) |
| 参考文献部分: | Keap1 facilitates p62-mediated ubiquitin aggregate clearance via autophagy. Fan W, Tang Z, Chen D, Moughon D, Ding X, Chen S, Zhu M, Zhong Q. Autophagy. 2010 Jul 22. 6(5):. 10.4161/auto.6.5.12189 PubMed 20495340 |
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 Flag-Keap1 7070 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Flag-Keap1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7070)
AUTHORS Fan W, Tang Z, Chen D, Moughon D, Ding X, Chen S, Zhu M, Zhong Q
TITLE Keap1 facilitates p62-mediated ubiquitin aggregate clearance via
autophagy.
JOURNAL Autophagy. 2010 Jul 22. 6(5):.
PUBMED 20495340
REFERENCE 2 (bases 1 to 7070)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7070
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 37..416
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 417..620
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 571..591
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 622..640
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 643..661
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 671..695
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 2718..2783
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
primer_bind complement(2824..2841)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2830..3054
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(3187..3206)
/label=F1ori-R
/note="F1 origin, reverse primer"
protein_bind 3338..3385
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 3393..4415
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
"
polyA_signal 4545..4666
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4582..4601)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4636..4655
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4715..4731)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4715..4731)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4728..4750)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4739..4755
/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(4763..4793)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4808..4829
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4946..4963)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5117..5705)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5197..5216)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5876..6736)
/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 6499..6518
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6737..6841)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6916..6935)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt
61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
421 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
481 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
541 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
601 aggtctatat aagcagagct ctccctatca gtgatagaga tctccctatc agtgatagag
661 atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
721 ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcg tttaaactta
781 agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattctgca gatatcatgc
841 agccagatcc caggcctagc ggggctgggg cctgctgccg attcctgccc ctgcagtcac
901 agtgccctga gggggcaggg gacgcggtga tgtacgcctc cactgagtgc aaggcggagg
961 tgacgccctc ccagcatggc aaccgcacct tcagctacac cctggaggat cataccaagc
1021 aggcctttgg catcatgaac gagctgcggc tcagccagca gctgtgtgac gtcacactgc
1081 aggtcaagta ccaggatgca ccggccgccc agttcatggc ccacaaggtg gtgctggcct
1141 catccagccc tgtcttcaag gccatgttca ccaacgggct gcgggagcag ggcatggagg
1201 tggtgtccat tgagggtatc caccccaagg tcatggagcg cctcattgaa ttcgcctaca
1261 cggcctccat ctccatgggc gagaagtgtg tcctccacgt catgaacggt gctgtcatgt
1321 accagatcga cagcgttgtc cgtgcctgca gtgacttcct ggtgcagcag ctggacccca
1381 gcaatgccat cggcatcgcc aacttcgctg agcagattgg ctgtgtggag ttgcaccagc
1441 gtgcccggga gtacatctac atgcattttg gggaggtggc caagcaagag gagttcttca
1501 acctgtccca ctgccaactg gtgaccctca tcagccggga cgacctgaac gtgcgctgcg
1561 agtccgaggt cttccacgcc tgcatcaact gggtcaagta cgactgcgaa cagcgacggt
1621 tctacgtcca ggcgctgctg cgggccgtgc gctgccactc gttgacgccg aacttcctgc
1681 agatgcagct gcagaagtgc gagatcctgc agtccgactc ccgctgcaag gactacctgg
1741 tcaagatctt cgaggagctc accctgcaca agcccacgca ggtgatgccc tgccgggcgc
1801 ccaaggtggg ccgcctgatc tacaccgcgg gcggctactt ccgacagtcg ctcagctacc
1861 tggaggctta caaccccagt gacggcacct ggctccggtt ggcggacctg caggtgccgc
1921 ggagcggcct ggccggctgc gtggtgggcg ggctgttgta cgccgtgggc ggcaggaaca
1981 actcgcccga cggcaacacc gactccagcg ccctggactg ttacaacccc atgaccaatc
2041 agtggtcgcc ctgcgccccc atgagcgtgc cccgtaaccg catcggggtg ggggtcatcg
2101 atggccacat ctatgccgtc ggcggctccc acggctgcat ccaccacaac agtgtggaga
2161 ggtatgagcc agagcgggat gagtggcact tggtggcccc aatgctgaca cgaaggatcg
2221 gggtgggcgt ggctgtcctc aatcgtctcc tttatgccgt ggggggcttt gacgggacaa
2281 accgccttaa ttcagctgag tgttactacc cagagaggaa cgagtggcga atgatcacag
2341 caatgaacac catccgaagc ggggcaggcg tctgcgtcct gcacaactgt atctatgctg
2401 ctgggggcta tgatggtcag gaccagctga acagcgtgga gcgctacgat gtggaaacag
2461 agacgtggac tttcgtagcc cccatgaagc accggcgaag tgccctgggg atcactgtcc
2521 accaggggag aatctacgtc cttggaggct atgatggtca cacgttcctg gacagtgtgg
2581 agtgttacga cccagataca gacacctgga gcgaggtgac ccgaatgaca tcgggccgga
2641 gtggggtggg cgtggctgtc accatggagc cctgccggaa gcagattgac cagcagaact
2701 gtacctgtgg cggccgcgac tacaaagacc atgacggtga ttataaagat catgacatcg
2761 actacaagga tgacgatgac aagtgactcg agtctagagg gcccgtttaa acccgctgat
2821 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
2881 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
2941 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
3001 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
3061 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
3121 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
3181 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
3241 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
3301 ccaaaaaact tgattagggt gatggttcac gtacctagaa gttcctattc cgaagttcct
3361 attctctaga aagtatagga acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg
3421 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
3481 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
3541 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg
3601 cgctcccgat tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca
3661 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
3721 ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga
3781 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
3841 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
3901 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
3961 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
4021 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
4081 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
4141 ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc
4201 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
4261 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
4321 gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
4381 gacgccccag cactcgtccg agggcaaagg aatagcacgt actacgagat ttcgattcca
4441 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
4501 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
4561 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
4621 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
4681 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
4741 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
4801 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
4861 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
4921 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
4981 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
5041 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
5101 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
5161 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
5221 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
5281 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
5341 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
5401 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
5461 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
5521 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
5581 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
5641 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
5701 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
5761 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
5821 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
5881 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
5941 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
6001 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
6061 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
6121 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
6181 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
6241 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
6301 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
6361 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
6421 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
6481 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
6541 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
6601 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
6661 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
6721 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
6781 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
6841 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
6901 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
6961 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
7021 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：pCgVchcrRNA
- 下一篇：hrGFP-WFY.m
