| 质粒名称: | pH3U3-mcs |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 12609 |
| 存储实验室: | Scot Wolfe |
|
载体骨架基本信息 |
|
| 载体名称: | pH3U3 |
| 载体出品公司: | Clontech |
| 空载体大小: | 5760bp |
| 完整质粒大小: | 5834bp |
| 载体类型: | 细菌表达 |
| 筛选标记: | URA3, HIS3 |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Multiple cloning site in His3 Ura3 reporter plasmid |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 44bp |
| GenBank编号 | DQ515893 |
| 启动子: | CMV |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NotI (not destroyed) |
| 3’克隆位点: | EcoRI (not destroyed) |
| 5’测序引物： | CAAATATGTATCCGCTCATGAC |
|
文献引用方法 |
|
| 材料和方法部分: | pH3U3-mcs was a gift from Scot Wolfe (Addgene plasmid # 12609 ; http://n2t.net/addgene:12609 ; RRID:Addgene_12609) |
| 参考文献部分: | A bacterial one-hybrid system for determining the DNA-binding specificity of transcription factors. Meng X, Brodsky MH, Wolfe SA. Nat Biotechnol. 2005 Aug . 23(8):988-94. 10.1038/nbt1120 PubMed 16041365 |
买家导航
pH3U3-mcs载体质粒基本信息
pH3U3-mcs质粒图谱载体图谱和pH3U3-mcs载体序列质粒序列多克隆位点信息
pH3U3-mcs质粒载体简介
pH3U3-mcs质粒序列载体序列
LOCUS pH3U3-mcs 5834 bp DNA circular SYN 25-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5834)
AUTHORS Meng X, Brodsky MH, Wolfe SA
TITLE A bacterial one-hybrid system for determining the DNA-binding
specificity of transcription factors.
JOURNAL Nat Biotechnol. 2005 Aug . 23(8):988-94.
PUBMED 16041365
REFERENCE 2 (bases 1 to 5834)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2005 Aug . 23(8):988-94."
FEATURES Location/Qualifiers
source 1..5834
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 12..98
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(633..1583)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 1631..1853
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
terminator complement(2345..2439)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(2470..3264)
/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(2581..2600)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 3191..3210
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
terminator 3419..3505
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3597..3624
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind 3702..3720
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3785..3815
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind 3825..3847
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3844..3860
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 3856..4518
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/label=HIS3
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
CDS 4566..5369
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
rep_origin complement(5378..5833)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5515..5536)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5727..5746
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 tctagaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
61 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgccctag acctagggta
121 cgggttttgc tgcccgcaaa cgggctgttc tggtgttgct agtttgttat cagaatcgca
181 gatccggctt caggtttgcc ggctgaaagc gctatttctt ccagaattgc catgattttt
241 tccccacggg aggcgtcact ggctcccgtg ttgtcggcag ctttgattcg ataagcagca
301 tcgcctgttt caggctgtct atgtgtgact gttgagctgt aacaagttgt ctcaggtgtt
361 caatttcatg ttctagttgc tttgttttac tggtttcacc tgttctatta ggtgttacat
421 gctgttcatc tgttacattg tcgatctgtt catggtgaac agctttaaat gcaccaaaaa
481 ctcgtaaaag ctctgatgta tctatctttt ttacaccgtt ttcatctgtg catatggaca
541 gttttccctt tgatatctaa cggtgaacag ttgttctact tttgtttgtt agtcttgatg
601 cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt agccagtatg
661 ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt gagatcatgc
721 ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt
781 aaagcatcgt gtagtgtttt tcttagtccg ttacgtaggt aggaatctga tgtaatggtt
841 gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt tacgagatcc
901 atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca
961 accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt caaaacccat
1021 tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa cttaaattca
1081 tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc ttttaataac
1141 cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc cagattatat
1201 tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa aactaattct
1261 aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa gcctttaacc
1321 aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt agctaataca
1381 ccataagcat tttccctact gatgttcatc atctgagcgt attggttata agtgaacgat
1441 accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc cacacagcat
1501 aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag ttcatttgct
1561 ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta atcactatac
1621 caattgagat gggctagtca atgataatta ctagtccttt tcctttgagt tgtgggtatc
1681 tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat
1741 tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata
1801 aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt
1861 tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac
1921 agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa atcgctgaat
1981 attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt
2041 cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg
2101 gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct tctcagggcg
2161 ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc
2221 ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta
2281 atgcacccag taaggcagcg gtatcatcaa caggcttacc cgtcttactg tccctagtgc
2341 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
2401 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc tcgaacccca
2461 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
2521 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
2581 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
2641 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
2701 atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc
2761 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
2821 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
2881 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
2941 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
3001 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
3061 cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac
3121 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
3181 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
3241 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatct
3301 tgatcccctg cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg
3361 cttcccaacc ttaccagagg gcgccccagc tggcaattcc gacgtcaccg gtaggcatca
3421 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
3481 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
3541 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
3601 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa
3661 tacattcaaa tatgtatccg ctcatgacat taacctataa aaataggcgt atcacgaggc
3721 cctttcgtct tcaaacgcgt gtacacccgg gcggccgcgt gcactccgga ggcgcgccga
3781 attctttaca ctttatgctt ccggctcgta tgttgtgtcg accgagcgga taacaatttc
3841 acacaggaaa cagctatgac agagcagaaa gccctagtaa agcgtattac aaatgaaacc
3901 aagattcaga ttgcgatctc tttaaagggt ggtcccctag cgatagagca ctcgatcttc
3961 ccagaaaaag aggcagaagc agtagcagaa caggccacac aatcgcaagt gattaacgtc
4021 cacacaggta tagggtttct ggaccatatg atacatgctc tggccaagca ttccggctgg
4081 tcgctaatcg ttgagtgcat tggtgactta cacatagacg accatcacac cactgaagac
4141 tgcgggattg ctctcggtca agcttttaaa gaggccctag gggccgtgcg tggagtaaaa
4201 aggtttggat caggatttgc gcctttggat gaggcacttt ccagagcggt ggtagatctt
4261 tcgaacaggc cgtacgcagt tgtcgaactt ggtttgcaaa gggagaaagt aggagatctc
4321 tcttgcgaga tgatcccgca ttttcttgaa agctttgcag aggctagcag aattaccctc
4381 cacgttgatt gtctgcgagg caagaatgat catcaccgta gtgagagtgc gttcaaggct
4441 cttgcggttg ccataagaga agccacctcg cccaatggta ccaacgatgt tccctccacc
4501 aaaggtgttc ttatgtagtg acaccgatta tttaaagctg cagggatccc acacaggaaa
4561 cagctatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt cctgttgctg
4621 ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc
4681 gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt tgtttactaa
4741 aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag ccgctaaagg
4801 cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct gacattggta
4861 atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg gcagacatta
4921 cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag gcggcggaag
4981 aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc aagggctccc
5041 tagctactgg agaatatact aagggtactg ttgacattgc gaagagcgac aaagattttg
5101 ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac gattggttga
5161 ttatgacacc cggtgtgggt ttagatgaca agggagacgc attgggtcaa cagtatagaa
5221 ccgtggatga tgtggtctct acaggatctg acattattat tgttggaaga ggactatttg
5281 caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc tgggaagcat
5341 atttgagaag atgcggccag caaaactaac tcgagctaaa ttgtaagcgt taatattttg
5401 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
5461 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
5521 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
5581 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
5641 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
5701 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
5761 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
5821 ctacagggcg cgtc
//
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