| 质粒名称: | BcatMutS33/S37.T41/S45 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 24204 |
| 存储实验室: | David Rimm |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3-2AB Flag |
| 载体出品公司: | -- |
| 空载体大小: | 5400 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄青霉素 |
| 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: | beta catenin |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 3000 bp |
| 基因突变: | S33A, S37A, T41A,S45A |
| GenBank编号 | -- |
| 启动子: | CMV |
| 标签/融合蛋白: | FLAG（插入件上的N端子） |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 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: | BcatMutS33/S37.T41/S45 was a gift from David Rimm (Addgene plasmid # 24204 ; http://n2t.net/addgene:24204 ; RRID:Addgene_24204) |
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 BcatMutS33/S37.T41/S45 7855 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS BcatMutS33/S37.T41/S45
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7855)
TITLE Rimm Plasmids
REFERENCE 2 (bases 1 to 7855)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7855
/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"
regulatory 894..903
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 903..926
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 957..3302
/codon_start=1
/gene="CTNNB1"
/product="protein involved both in cell-cell adhesion as
part of the cadherin protein complex, and in gene
transcription as part of the Wnt signaling pathway"
/label=CTNNB1
/note="also known as catenin beta-1; related to Drosophila
armadillo"
/translation="MATQADLMELDMAMEPDRKAAVSHWQQQSYLDAGIHAGATATAPA
LSGKGNPEEEDVDTSQVLYEWEQGFSQSFTQEQVADIDGQYAMTRAQRVRAAMFPETLD
EGMQIPSTQFDAAHPTNVQRLAEPSQMLKHAVVNLINYQDDAELATRAIPELTKLLNDE
DQVVVNKAAVMVHQLSKKEASRHAIMRSPQMVSAIVRTMQNTNDVETARCTAGTLHNLS
HHREGLLAIFKSGGIPALVKMLGSPVDSVLFYAITTLHNLLLHQEGAKMAVRLAGGLQK
MVALLNKTNVKFLAITTDCLQILAYGNQESKLIILASGGPQALVNIMRTYTYEKLLWTT
SRVLKVLSVCSSNKPAIVEAGGMQALGLHLTDPSQRLVQNCLWTLRNLSDAATKQEGME
GLLGTLVQLLGSDDINVVTCAAGILSNLTCNNYKNKMMVCQVGGIEALVRTVLRAGDRE
DITEPAICALRHLTSRHQEAEMAQNAVRLHYGLPVVVKLLHPPSHWPLIKATVGLIRNL
ALCPANHAPLREQGAIPRLVQLLVRAHQDTQRRTSMGGTQQQFVEGVRMEEIVEGCTGA
LHILARDVHNRIVIRGLNTIPLFVQLLYSPIENIQRVAAGVLCELAQDKEAAEAIEAEG
ATAPLTELLHSRNEGVATYAAAVLFRMSEDKPQDYKKRLSVELTSSLFRTEPMAWNETA
DLGLDIGAQGEPLGYRQDDPSYRSFHSGGYGQDALGMDPMMEHEMGGHHPGADYPVDGL
PDLGHAQDLMDGLPPGDSNQLAWFDTDL"
promoter complement(3408..3426)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3409..3426)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(3446..3463)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3452..3676
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3722..4150
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3809..3828)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4019..4040
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4159..4179)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4164..4493
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4344..4479
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4406..4425
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4560..5354
/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(4614..4633)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5224..5243
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5528..5649
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5565..5584)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5619..5638
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5698..5714)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5698..5714)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5711..5733)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5722..5738
/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(5746..5776)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5791..5812
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5929..5946)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6100..6688)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6180..6199)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6859..7719)
/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 7482..7501
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7720..7824)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctgccacca
901 tggactacaa ggacgacgat gacaagggac ctaagcttgg taccgagctc ggatccatgg
961 ctactcaagc tgatttgatg gagttggaca tggccatgga accagacaga aaagcggctg
1021 ttagtcactg gcagcaacag tcttacctgg acgctggaat ccatgctggt gccactgcca
1081 cagctcctgc tctgagtggt aaaggcaatc ctgaggaaga ggatgtggat acctcccaag
1141 tcctgtatga gtgggaacag ggattttctc agtccttcac tcaagaacaa gtagctgata
1201 ttgatggaca gtatgcaatg actcgagctc agagggtacg agctgctatg ttccctgaga
1261 cattagatga gggcatgcag atcccatcta cacagtttga tgctgctcat cccactaatg
1321 tccagcgttt ggctgaacca tcacagatgc tgaaacatgc agttgtaaac ttgattaact
1381 atcaagatga tgcagaactt gccacacgtg caatccctga actgacaaaa ctgctaaatg
1441 acgaggacca ggtggtggtt aataaggctg cagttatggt ccatcagctt tctaaaaagg
1501 aagcttccag acacgctatc atgcgttctc ctcagatggt gtctgctatt gtacgtacca
1561 tgcagaatac aaatgatgta gaaacagctc gttgtaccgc tgggaccttg cataaccttt
1621 cccatcatcg tgagggctta ctggccatct ttaagtctgg aggcattcct gccctggtga
1681 aaatgcttgg ttcaccagtg gattctgtgt tgttttatgc cattacaact ctccacaacc
1741 ttttattaca tcaagaagga gctaaaatgg cagtgcgttt agctggtggg ctgcagaaaa
1801 tggttgcctt gctcaacaaa acaaatgtta aattcttggc tattacgaca gactgccttc
1861 aaattttagc ttatggcaac caagaaagca agctcatcat actggctagt ggtggacccc
1921 aagctttagt aaatataatg aggacctata cttacgaaaa actactgtgg accacaagca
1981 gagtgctgaa ggtgctatct gtctgctcta gtaataagcc ggctattgta gaagctggtg
2041 gaatgcaagc tttaggactt cacctgacag atccaagtca acgtcttgtt cagaactgtc
2101 tttggactct caggaatctt tcagatgctg caactaaaca ggaagggatg gaaggtctcc
2161 ttgggactct tgttcagctt ctgggttcag atgatataaa tgtggtcacc tgtgcagctg
2221 gaattctttc taacctcact tgcaataatt ataagaacaa gatgatggtc tgccaagtgg
2281 gtggtataga ggctcttgtg cgtactgtcc ttcgggctgg tgacagggaa gacatcactg
2341 agcctgccat ctgtgctctt cgtcatctga ccagccgaca ccaagaagca gagatggccc
2401 agaatgcagt tcgccttcac tatggactac cagttgtggt taagctctta cacccaccat
2461 cccactggcc tctgataaag gctactgttg gattgattcg aaatcttgcc ctttgtcccg
2521 caaatcatgc acctttgcgt gagcagggtg ccattccacg actagttcag ttgcttgttc
2581 gtgcacatca ggatacccag cgccgtacgt ccatgggtgg gacacagcag caatttgtgg
2641 agggggtccg catggaagaa atagttgaag gttgtaccgg agcccttcac atcctagctc
2701 gggatgttca caaccgaatt gttatcagag gactaaatac cattccattg tttgtgcagc
2761 tgctttattc tcccattgaa aacatccaaa gagtagctgc aggggtcctc tgtgaacttg
2821 ctcaggacaa ggaagctgca gaagctattg aagctgaggg agccacagct cctctgacag
2881 agttacttca ctctaggaat gaaggtgtgg cgacatatgc agctgctgtt ttgttccgaa
2941 tgtctgagga caagccacaa gattacaaga aacggctttc agttgagctg accagctctc
3001 tcttcagaac agagccaatg gcttggaatg agactgctga tcttggactt gatattggtg
3061 cccagggaga accccttgga tatcgccagg atgatcctag ctatcgttct tttcactctg
3121 gtggatatgg ccaggatgcc ttgggtatgg accccatgat ggaacatgag atgggtggcc
3181 accaccctgg tgctgactat ccagttgatg ggctgccaga tctggggcat gcccaggacc
3241 tcatggatgg gctgcctcca ggtgacagca atcagctggc ctggtttgat actgacctgt
3301 aaatcatcct ttaggaggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat
3361 atccatcaca ctggcggccg ctcgagcatg catctagagg gccctattct atagtgtcac
3421 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
3481 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
3541 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
3601 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctgggaat
3661 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
3721 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
3781 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
3841 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
3901 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
3961 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
4021 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
4081 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
4141 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
4201 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
4261 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
4321 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
4381 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
4441 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
4501 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
4561 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
4621 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
4681 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
4741 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
4801 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
4861 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
4921 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
4981 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
5041 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
5101 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
5161 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
5221 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
5281 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
5341 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
5401 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
5461 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
5521 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
5581 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
5641 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
5701 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
5761 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
5821 gcgctcactg cccgctttcc agttgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
5881 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
5941 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
6001 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
6061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
6121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
6181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
6241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
6301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
6361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
6421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
6481 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
6541 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
6601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
6661 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
6721 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
6781 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
6841 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
6901 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
6961 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
7021 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
7081 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
7141 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
7201 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
7261 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
7321 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
7381 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
7441 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
7501 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
7561 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
7621 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
7681 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
7741 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
7801 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
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