| 质粒名称: | pT3-EF1aH N90-beta-catenin |
|---|---|
| 目录编号: | 86499 |
| 存储实验室: | Xin Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pT3-EF1aH |
| 完整质粒大小: | 7299 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: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: | beta-catenin /b-catenin /CTNNB1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 2121 bp |
| 基因突变: | Deletion of amino acids 1-90; constitutively active |
| 启动子: | EF1a |
| 标签/融合蛋白: | Myc (N terminal on backbone) |
| 克隆方法: | Gateway Cloning |
| 5’测序引物 | EF1a-F |
|
文献引用方法 |
|
| 材料和方法部分: | pT3-EF1aH N90-beta-catenin was a gift from Xin Chen (Addgene plasmid # 86499 ; http://n2t.net/addgene:86499 ; RRID:Addgene_86499) |
| 参考文献部分: | Distinct pathways of genomic progression to benign and malignant tumors of the liver. Tward AD, Jones KD, Yant S, Cheung ST, Fan ST, Chen X, Kay MA, Wang R, Bishop JM. Proc Natl Acad Sci U S A. 2007 Sep 11;104(37):14771-6. Epub 2007 Sep 4. 10.1073/pnas.0706578104 PubMed 17785413 |
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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-EF1aH N90-beta-catenin 7299 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of Myc tagged N90 beta-catenin.
ACCESSION .
VERSION .
KEYWORDS pT3-EF1aH N90-beta-catenin
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7299)
AUTHORS Tward AD, Jones KD, Yant S, Cheung ST, Fan ST, Chen X, Kay MA, Wang
R, Bishop JM
TITLE Distinct pathways of genomic progression to benign and malignant
tumors of the liver.
JOURNAL Proc Natl Acad Sci U S A. 2007 Sep 11;104(37):14771-6. Epub 2007 Sep
4.
PUBMED 17785413
REFERENCE 2 (bases 1 to 7299)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7299
/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"
protein_bind 1210..1234
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 1265..1294
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
protein_bind complement(3406..3430)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind 3458..3477
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3512..3529)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3518..3742
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(4160..4176)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4160..4176)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4173..4195)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4184..4200
/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(4208..4238)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4253..4274
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4391..4408)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4562..5150)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4642..4661)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5321..6181)
/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 5944..5963
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6182..6286)
/gene="bla"
/label=AmpR promoter
primer_bind 6354..6372
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(6410..6432)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 6532..6551
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6745..6767
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6759..6776
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6760..6776
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7158..7178)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
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 ggtaccaata
1201 cggatatcaa caagtttgta caaaaaagca ggctttaaag gaaccaattc agtcgaccac
1261 catggagcag aagctgatct ccgaggagga cctgaaggat cctcgagctc agagggtacg
1321 agctgctatg ttccctgaga cattagatga gggcatgcag atcccatcta cacagtttga
1381 tgctgctcat cccactaatg tccagcgttt ggctgaacca tcacagatgc tgaaacatgc
1441 agttgtaaac ttgattaact atcaagatga tgcagaactt gccacacgtg caatccctga
1501 actgacaaaa ctgctaaatg acgaggacca ggtggtggtt aataaggctg cagttatggt
1561 ccatcagctt tctaaaaagg aagcttccag acacgctatc atgcgttctc ctcagatggt
1621 gtctgctatt gtacgtacca tgcagaatac aaatgatgta gaaacagctc gttgtaccgc
1681 tgggaccttg cataaccttt cccatcatcg tgagggctta ctggccatct ttaagtctgg
1741 aggcattcct gccctggtga aaatgcttgg ttcaccagtg gattctgtgt tgttttatgc
1801 cattacaact ctccacaacc ttttattaca tcaagaagga gctaaaatgg cagtgcgttt
1861 agctggtggg ctgcagaaaa tggttgcctt gctcaacaaa acaaatgtta aattcttggc
1921 tattacgaca gactgccttc aaattttagc ttatggcaac caagaaagca agctcatcat
1981 actggctagt ggtggacccc aagctttagt aaatataatg aggacctata cttacgaaaa
2041 actactgtgg accacaagca gagtgctgaa ggtgctatct gtctgctcta gtaataagcc
2101 ggctattgta gaagctggtg gaatgcaagc tttaggactt cacctgacag atccaagtca
2161 acgtcttgtt cagaactgtc tttggactct caggaatctt tcagatgctg caactaaaca
2221 ggaagggatg gaaggtctcc ttgggactct tgttcagctt ctgggttcag atgatataaa
2281 tgtggtcacc tgtgcagctg gaattctttc taacctcact tgcaataatt ataagaacaa
2341 gatgatggtc tgccaagtgg gtggtataga ggctcttgtg cgtactgtcc ttcgggctgg
2401 tgacagggaa gacatcactg agcctgccat ctgtgctctt cgtcatctga ccagccgaca
2461 ccaagaagca gagatggccc agaatgcagt tcgccttcac tatggactac cagttgtggt
2521 taagctctta cacccaccat cccactggcc tctgataaag gctactgttg gattgattcg
2581 aaatcttgcc ctttgtcccg caaatcatgc acctttgcgt gagcagggtg ccattccacg
2641 actagttcag ttgcttgttc gtgcacatca ggatacccag cgccgtacgt ccatgggtgg
2701 gacacagcag caatttgtgg agggggtccg catggaagaa atagttgaag gttgtaccgg
2761 agcccttcac atcctagctc gggatgttca caaccgaatt gttatcagag gactaaatac
2821 cattccattg tttgtgcagc tgctttattc tcccattgaa aacatccaaa gagtagctgc
2881 aggggtcctc tgtgaacttg ctcaggacaa ggaagctgca gaagctattg aagctgaggg
2941 agccacagct cctctgacag agttacttca ctctaggaat gaaggtgtgg cgacatatgc
3001 agctgctgtt ttgttccgaa tgtctgagga caagccacaa gattacaaga aacggctttc
3061 agttgagctg accagctctc tcttcagaac agagccaatg gcttggaatg agactgctga
3121 tcttggactt gatattggtg cccagggaga accccttgga tatcgccagg atgatcctag
3181 ctatcgttct tttcactctg gtggatatgg ccaggatgcc ttgggtatgg accccatgat
3241 ggaacatgag atgggtggcc accaccctgg tgctgactat ccagttgatg ggctgccaga
3301 tctggggcat gcccaggacc tcatggatgg gctgcctcca ggtgacagca atcagctggc
3361 ctggtttgat actgacctgt aagcggccgc actcgagata tctagaccca gctttcttgt
3421 acaaagtggt tgatatcgcg gccgctgcat tctagttgtg gtttgtccaa actcatcaat
3481 gtatcttatc atgtctggat ccgctgatca gcctcgactg tgccttctag ttgccagcca
3541 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
3601 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
3661 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
3721 ggggatgcgg tgggctctat ggcttctgag gcgggatccc cttgaaatac atccacaggt
3781 acacctccaa ttgactcaaa tgatgtcaat tagtctatca gaagcttcta aagccatgac
3841 atcattttct ggaattttcc aagctgttta aaggcacagt caacttagtg tatgtaaact
3901 tctgacccac tggaattgtg atacagtgaa ttataagtga aataatctgt ctgtaaacaa
3961 ttgttggaaa aatgacttgt gtcatgcaca aagtagatgt cctaactgac ttgccaaaac
4021 tattgtttgt taacaagaaa tttgtggagt agttgaaaaa cgagttttaa tgactccaac
4081 ttaagtgtat gtaaacttcc gacttcaact gtataggtct agagtcgacc tgcaggcatg
4141 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
4201 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
4261 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
4321 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
4381 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
4441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
4501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
4561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
4621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
4681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
4741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
4801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
4861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
4921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
4981 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
5041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
5101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
5161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
5221 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
5281 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
5341 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
5401 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
5461 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
5521 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
5581 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
5641 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
5701 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
5761 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
5821 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
5881 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
5941 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
6001 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
6061 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
6121 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
6181 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
6241 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
6301 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
6361 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
6421 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
6481 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
6541 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
6601 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
6661 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
6721 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
6781 tcgagctcgg taccctacag ttgaagtcgg aagtttacat acacttaagt tggagtcatt
6841 aaaactcgtt tttcaactac tccacaaatt tcttgttaac aaacaatagt tttggcaagt
6901 cagttaggac atctactttg tgcatgacac aagtcatttt tccaacaatt gtttacagac
6961 agattatttc acttataatt cactgtatca caattccagt gggtcagaag tttacataca
7021 ctaagttgac tgtgccttta aacagcttgg aaaattccag aaaatgatgt catggcttta
7081 gaagcttctg atagactaat tgacatcatt tgagtcaatt ggaggtgtac ctgtggatgt
7141 atttcaagga attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
7201 agcaggcaga agtatgcaaa gcatgcatat cgatactagt ttaattaaga tcctctttgc
7261 agctaatgga ccttctaggt cttgaaagga gtgggaatt
//
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