| 质粒名称: | pT3-myr-AKT-HA |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31789 |
| 存储实验室: | Xin Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pT3-EF1a |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 7000 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Akt1 |
| 物种来源: | -- |
| 插入大小: | 1503bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pT3-myr-AKT-HA was a gift from Xin Chen (Addgene plasmid # 31789 ; http://n2t.net/addgene:31789 ; RRID:Addgene_31789) |
| 参考文献部分: | Increased lipogenesis, induced by AKT-mTORC1-RPS6 signaling, promotes development of human hepatocellular carcinoma. Calvisi DF, Wang C, Ho C, Ladu S, Lee SA, Mattu S, Destefanis G, Delogu S, Zimmermann A, Ericsson J, Brozzetti S, Staniscia T, Chen X, Dombrowski F, Evert M. Gastroenterology. 2011 Mar;140(3):1071-83. Epub 2010 Dec 11. 10.1053/j.gastro.2010.12.006 PubMed 21147110 |
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 pT3-myr-AKT-HA 7091 bp ds-DNA circular SYN 29-JUL-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pT3-myr-AKT-HA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7091)
AUTHORS Calvisi DF, Wang C, Ho C, Ladu S, Lee SA, Mattu S, Destefanis G,
Delogu S, Zimmermann A, Ericsson J, Brozzetti S, Staniscia T, Chen
X, Dombrowski F, Evert M
TITLE Increased lipogenesis, induced by AKT-mTORC1-RPS6 signaling,
promotes development of human hepatocellular carcinoma.
JOURNAL Gastroenterology. 2011 Mar;140(3):1071-83. Epub 2010 Dec 11.
PUBMED 21147110
REFERENCE 2 (bases 1 to 7091)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 29, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7091
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..1179
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 232..1170
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1127..1147
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
primer_bind 1196..1215
/label=T7
/note="T7 promoter, forward primer"
promoter 1196..1214
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1293..1317
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2811..2837
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
protein_bind complement(2857..2881)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 2934..2975
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
primer_bind complement(3013..3030)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3019..3243
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
protein_bind complement(3313..3346)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind 3402..3422
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
primer_bind complement(3804..3821)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3804..3820)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3813..3835)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(4029..4048)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4148..4170
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4208..4226)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4294..4398
/gene="bla"
/label=AmpR promoter
CDS 4399..5259
/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 complement(4617..4636)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5430..6018
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5919..5938
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6172..6189
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6306..6327
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6342..6372
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6380..6396
/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)."
primer_bind 6385..6407
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6404..6420
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6404..6420
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6846..6865)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
protein_bind complement(6918..6951)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
ORIGIN
1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggatccac
1261 tagtccagtg tggtggaatt ctgcagatat caacaagttt gtacaaaaaa gcaggctccg
1321 cggccgcccc cttcaccatg gggagcagca agagcaagcc caagtctaga atgaacgacg
1381 tagccattgt gaaggagggc tggctgcaca aacgagggga atatattaaa acctggcggc
1441 cacgctactt cctcctcaag aacgatggca cctttattgg ctacaaggaa cggcctcagg
1501 atgtggatca gcgagagtcc ccactcaaca acttctcagt ggcacaatgc cagctgatga
1561 agacagagcg gccaaggccc aacaccttta tcatccgctg cctgcagtgg accacagtca
1621 ttgagcgcac cttccatgtg gaaacgcctg aggagcggga agaatgggcc accgccattc
1681 agactgtggc cgatggactc aagaggcagg aagaagagac gatggacttc cgatcaggct
1741 cacccagtga caactcaggg gctgaagaga tggaggtgtc cctggccaag cccaagcacc
1801 gtgtgaccat gaacgagttt gagtacctga aactactggg caagggcacc tttgggaaag
1861 tgattctggt gaaagagaag gccacaggcc gctactatgc catgaagatc ctcaagaagg
1921 aggtcatcgt cgccaaggat gaggttgccc acacgcttac tgagaaccgt gtcctgcaga
1981 actctaggca tcccttcctt acggccctca agtactcatt ccagacccac gaccgcctct
2041 gctttgtcat ggagtatgcc aacgggggcg agctcttctt ccacctgtct cgagagcgcg
2101 tgttctccga ggaccgggcc cgcttctatg gtgcggagat tgtgtctgcc ctggactact
2161 tgcactccga gaagaacgtg gtgtaccggg acctgaagct ggagaacctc atgctggaca
2221 aggacgggca catcaagata acggacttcg ggctgtgcaa ggaggggatc aaggatggtg
2281 ccactatgaa gacattctgc ggaacgccgg agtacctggc ccctgaggtg ctggaggaca
2341 acgactacgg ccgtgcagtg gactggtggg ggctgggcgt ggtcatgtat gagatgatgt
2401 gtggccgcct gcccttctac aaccaggacc acgagaagct gttcgagctg atcctcatgg
2461 aggagatccg cttcccgcgc acactcggcc ctgaggccaa gtccctgctc tccgggctgc
2521 tcaagaagga ccctacacag aggctcggtg ggggctctga ggatgccaag gagatcatgc
2581 agcaccggtt ctttgccaac atcgtgtggc aggatgtgta tgagaagaag ctgagcccac
2641 ctttcaagcc ccaggtcacc tctgagactg acaccaggta tttcgatgag gagttcacag
2701 ctcagatgat caccatcacg ccgcctgatc aagatgacag catggagtgt gtggacagtg
2761 agcggaggcc gcacttcccc cagttctcct actcagccag tggcacagcc tacccatacg
2821 acgtcccaga ctacgcttga aagggtgggc gcgccgaccc agctttcttg tacaaagtgg
2881 ttgatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc
2941 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa tgagtttaaa
3001 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
3061 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
3121 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
3181 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
3241 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
3301 gcggcgcatt aaataacttc gtataatgta tgctatacga agttatatcg tatgcatgct
3361 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
3421 ccacagaatt ccttgaaata catccacagg tacacctcca attgactcaa atgatgtcaa
3481 ttagtctatc agaagcttct aaagccatga catcattttc tggaattttc caagctgttt
3541 aaaggcacag tcaacttagt gtatgtaaac ttctgaccca ctggaattgt gatacagtga
3601 attataagtg aaataatctg tctgtaaaca attgttggaa aaatgacttg tgtcatgcac
3661 aaagtagatg tcctaactga cttgccaaaa ctattgtttg ttaacaagaa atttgtggag
3721 tagttgaaaa acgagtttta atgactccaa cttaagtgta tgtaaacttc cgacttcaac
3781 tgtagggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
3841 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
3901 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
3961 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
4021 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
4081 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
4141 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
4201 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
4261 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
4321 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
4381 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
4441 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
4501 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
4561 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
4621 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
4681 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
4741 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
4801 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
4861 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
4921 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
4981 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
5041 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
5101 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
5161 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
5221 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
5281 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
5341 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
5401 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
5461 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
5521 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
5581 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
5641 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
5701 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
5761 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
5821 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
5881 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
5941 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
6001 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
6061 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
6121 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
6181 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
6241 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
6301 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
6361 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
6421 catgattacg ccaagcttgc atgcctgcag gtcgactcta gacctataca gttgaagtcg
6481 gaagtttaca tacacttaag ttggagtcat taaaactcgt ttttcaacta ctccacaaat
6541 ttcttgttaa caaacaatag ttttggcaag tcagttagga catctacttt gtgcatgaca
6601 caagtcattt ttccaacaat tgtttacaga cagattattt cacttataat tcactgtatc
6661 acaattccag tgggtcagaa gtttacatac actaagttga ctgtgccttt aaacagcttg
6721 gaaaattcca gaaaatgatg tcatggcttt agaagcttct gatagactaa ttgacatcat
6781 ttgagtcaat tggaggtgta cctgtggatg tatttcaagg ggatccagac atgataagat
6841 acattgatga gtttggacaa accacaacta gaatgcagcg gccgctagct cgagatctta
6901 attaaactag tatcgatata acttcgtata atgtatgcta tacgaagtta tatgtacggg
6961 ccagatatac gcgttgacat tgattattga ctaggctttt gcaaaaagct ttgcaaagat
7021 ggataaagtt ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag
7081 gagtgggaat t
//
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