| 质粒名称: | pT3EF1aH-myr-Akt |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 179909 |
| 存储实验室: | Xin Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pT3EF1aH |
| 空载体大小: | 7000bp |
| 完整质粒大小: | 6694 bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | Akt1 / Akt |
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 1503 bp |
| GenBank编号 | NM_001165894 |
| 标签/融合蛋白: |
Myr (N terminal on backbone)
HA (C terminal on insert)
|
| 克隆方法: | Gateway Cloning |
| 5’测序引物 | EF-1a-Fwd |
| 3’测序引物 | BGH Reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pT3EF1aH-myr-Akt was a gift from Xin Chen (Addgene plasmid # 179909 ; http://n2t.net/addgene:179909 ; RRID:Addgene_179909) |
| 参考文献部分: | AKT (v-akt murine thymoma viral oncogene homolog 1) and N-Ras (neuroblastoma ras viral oncogene homolog) coactivation in the mouse liver promotes rapid carcinogenesis by way of mTOR (mammalian target of rapamycin complex 1), FOXM1 (forkhead box M1)/SKP2, and c-Myc pathways. Ho C, Wang C, Mattu S, Destefanis G, Ladu S, Delogu S, Armbruster J, Fan L, Lee SA, Jiang L, Dombrowski F, Evert M, Chen X, Calvisi DF. Hepatology. 2012 Mar;55(3):833-45. doi: 10.1002/hep.24736. Epub 2011 Dec 19. 10.1002/hep.24736 PubMed 21993994 |
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 pT3EF1aH-myr-Akt 6694 bp ds-DNA circular SYN 25-MAR-2022
DEFINITION This plasmid is in the pT3-EF1a vector without loxP sites flanking
the inverted repeats of SB (sleeping beauty) sequence. Therefore
this plasmid can be used with Cre for in vivo studies..
ACCESSION .
VERSION .
KEYWORDS pT3EF1aH-myr-Akt
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6694)
AUTHORS Ho C, Wang C, Mattu S, Destefanis G, Ladu S, Delogu S, Armbruster J,
Fan L, Lee SA, Jiang L, Dombrowski F, Evert M, Chen X, Calvisi DF
TITLE AKT (v-akt murine thymoma viral oncogene homolog 1) and N-Ras
(neuroblastoma ras viral oncogene homolog) coactivation in the mouse
liver promotes rapid carcinogenesis by way of mTOR (mammalian target
of rapamycin complex 1), FOXM1 (forkhead box M1)/SKP2, and c-Myc
pathways.
JOURNAL Hepatology. 2012 Mar;55(3):833-45. doi: 10.1002/hep.24736. Epub 2011
Dec 19.
PUBMED 21993994
REFERENCE 2 (bases 1 to 6694)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 25, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6694
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(35..52)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(206..794)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(286..305)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(965..1825)
/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 1588..1607
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1826..1930)
/gene="bla"
/label=AmpR promoter
primer_bind 1998..2016
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2054..2076)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2176..2195
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2389..2411
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2403..2420
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2404..2420
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2802..2822)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2944..4122
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 3175..4113
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 4070..4090
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
protein_bind 4153..4177
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 4212..4232
/codon_start=1
/product="N-myristoylation signal from Src kinase (Pellman
et al., 1985; Kaplan et al., 1988)"
/label=myr
/translation="MGSSKSK"
CDS 5685..5711
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
protein_bind complement(5744..5768)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind 5796..5815
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5850..5867)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5856..6080
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6498..6514)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6498..6514)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6511..6533)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6522..6538
/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(6546..6576)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6591..6612
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
1921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
1981 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
2041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
2101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
2161 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
2221 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
2281 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
2341 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
2401 gttgtaaaac gacggccagt gaattcgagc tcggtaccct acagttgaag tcggaagttt
2461 acatacactt aagttggagt cattaaaact cgtttttcaa ctactccaca aatttcttgt
2521 taacaaacaa tagttttggc aagtcagtta ggacatctac tttgtgcatg acacaagtca
2581 tttttccaac aattgtttac agacagatta tttcacttat aattcactgt atcacaattc
2641 cagtgggtca gaagtttaca tacactaagt tgactgtgcc tttaaacagc ttggaaaatt
2701 ccagaaaatg atgtcatggc tttagaagct tctgatagac taattgacat catttgagtc
2761 aattggaggt gtacctgtgg atgtatttca aggaattctg tggaatgtgt gtcagttagg
2821 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atatcgatac
2881 tagtttaatt aagatcctct ttgcagctaa tggaccttct aggtcttgaa aggagtggga
2941 attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
3001 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
3061 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
3121 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
3181 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
3241 attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
3301 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
3361 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
3421 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
3481 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
3541 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
3601 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
3661 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
3721 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
3781 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
3841 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
3901 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
3961 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
4021 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
4081 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggaattag cttggtacca
4141 atacggatat caacaagttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
4201 tggatcccac catggggagc agcaagagca agcccaagtc tagaatgaac gacgtagcca
4261 ttgtgaagga gggctggctg cacaaacgag gggaatatat taaaacctgg cggccacgct
4321 acttcctcct caagaacgat ggcaccttta ttggctacaa ggaacggcct caggatgtgg
4381 atcagcgaga gtccccactc aacaacttct cagtggcaca atgccagctg atgaagacag
4441 agcggccaag gcccaacacc tttatcatcc gctgcctgca gtggaccaca gtcattgagc
4501 gcaccttcca tgtggaaacg cctgaggagc gggaagaatg ggccaccgcc attcagactg
4561 tggccgatgg actcaagagg caggaagaag agacgatgga cttccgatca ggctcaccca
4621 gtgacaactc aggggctgaa gagatggagg tgtccctggc caagcccaag caccgtgtga
4681 ccatgaacga gtttgagtac ctgaaactac tgggcaaggg cacctttggg aaagtgattc
4741 tggtgaaaga gaaggccaca ggccgctact atgccatgaa gatcctcaag aaggaggtca
4801 tcgtcgccaa ggatgaggtt gcccacacgc ttactgagaa ccgtgtcctg cagaactcta
4861 ggcatccctt ccttacggcc ctcaagtact cattccagac ccacgaccgc ctctgctttg
4921 tcatggagta tgccaacggg ggcgagctct tcttccacct gtctcgagag cgcgtgttct
4981 ccgaggaccg ggcccgcttc tatggtgcgg agattgtgtc tgccctggac tacttgcact
5041 ccgagaagaa cgtggtgtac cgggacctga agctggagaa cctcatgctg gacaaggacg
5101 ggcacatcaa gataacggac ttcgggctgt gcaaggaggg gatcaaggat ggtgccacta
5161 tgaagacatt ctgcggaacg ccggagtacc tggcccctga ggtgctggag gacaacgact
5221 acggccgtgc agtggactgg tgggggctgg gcgtggtcat gtatgagatg atgtgtggcc
5281 gcctgccctt ctacaaccag gaccacgaga agctgttcga gctgatcctc atggaggaga
5341 tccgcttccc gcgcacactc ggccctgagg ccaagtccct gctctccggg ctgctcaaga
5401 aggaccctac acagaggctc ggtgggggct ctgaggatgc caaggagatc atgcagcacc
5461 ggttctttgc caacatcgtg tggcaggatg tgtatgagaa gaagctgagc ccacctttca
5521 agccccaggt cacctctgag actgacacca ggtatttcga tgaggagttc acagctcaga
5581 tgatcaccat cacgccgcct gatcaagatg acagcatgga gtgtgtggac agtgagcgga
5641 ggccgcactt cccccagttc tcctactcag ccagtggcac agcctaccca tacgacgtcc
5701 cagactacgc ttgagaattc gcggccgcac tcgagatatc tagacccagc tttcttgtac
5761 aaagtggttg atatcgcggc cgctgcattc tagttgtggt ttgtccaaac tcatcaatgt
5821 atcttatcat gtctggatcc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
5881 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
5941 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
6001 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
6061 ggatgcggtg ggctctatgg cttctgaggc gggatcccct tgaaatacat ccacaggtac
6121 acctccaatt gactcaaatg atgtcaatta gtctatcaga agcttctaaa gccatgacat
6181 cattttctgg aattttccaa gctgtttaaa ggcacagtca acttagtgta tgtaaacttc
6241 tgacccactg gaattgtgat acagtgaatt ataagtgaaa taatctgtct gtaaacaatt
6301 gttggaaaaa tgacttgtgt catgcacaaa gtagatgtcc taactgactt gccaaaacta
6361 ttgtttgtta acaagaaatt tgtggagtag ttgaaaaacg agttttaatg actccaactt
6421 aagtgtatgt aaacttccga cttcaactgt ataggtctag agtcgacctg caggcatgca
6481 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
6541 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
6601 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
6661 cagctgcatt aatgaatcgg ccaacgcgcg ggga
//
