| 质粒名称: | pH3U3-Zif268 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 12610 |
| 存储实验室: | Scot Wolfe |
|
载体骨架基本信息 |
|
| 载体名称: | PH3U3-mcs |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 5821 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | URA3, HIS3 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Zif268 binding site in pH3U3 |
| 物种来源: | -- |
| 插入大小: | 21 bp |
| 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: | AscI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CAAATATGTATCCGCTCATGAC |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pH3U3-Zif268 was a gift from Scot Wolfe (Addgene plasmid # 12610 ; http://n2t.net/addgene:12610 ; RRID:Addgene_12610) |
| 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
pH3U3Zif268载体基本信息
pH3U3Zif268载体图谱质粒图谱和多克隆位点信息
pH3U3Zif268载体简介
pH3U3Zif268载体序列
LOCUS pH3U3-Zif268 5842 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pH3U3-Zif268
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5842)
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 5842)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5842
/organism="synthetic DNA construct"
/mol_type="other DNA"
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 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 3793..3823
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind 3833..3855
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3852..3868
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3852..3868
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS 3864..4526
/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 4574..5377
/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(5386..5841)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5523..5544)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5735..5754
/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 gcggccgctg cgtgggcggc actccggagg
3781 cgcgccgaat tctttacact ttatgcttcc ggctcgtatg ttgtgtcgac cgagcggata
3841 acaatttcac acaggaaaca gctatgacag agcagaaagc cctagtaaag cgtattacaa
3901 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
3961 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
4021 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
4081 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
4141 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctaggg gccgtgcgtg
4201 gagtaaaaag gtttggatca ggatttgcgc ctttggatga ggcactttcc agagcggtgg
4261 tagatctttc gaacaggccg tacgcagttg tcgaacttgg tttgcaaagg gagaaagtag
4321 gagatctctc ttgcgagatg atcccgcatt ttcttgaaag ctttgcagag gctagcagaa
4381 ttaccctcca cgttgattgt ctgcgaggca agaatgatca tcaccgtagt gagagtgcgt
4441 tcaaggctct tgcggttgcc ataagagaag ccacctcgcc caatggtacc aacgatgttc
4501 cctccaccaa aggtgttctt atgtagtgac accgattatt taaagctgca gggatcccac
4561 acaggaaaca gctatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc
4621 tgttgctgcc aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt
4681 ggatgttcgt accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg
4741 tttactaaaa acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc
4801 gctaaaggca ttatccgcca agtacaattt tttactcttc gaagacagaa aatttgctga
4861 cattggtaat acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc
4921 agacattacg aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc
4981 ggcggaagaa gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa
5041 gggctcccta gctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa
5101 agattttgtt atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga
5161 ttggttgatt atgacacccg gtgtgggttt agatgacaag ggagacgcat tgggtcaaca
5221 gtatagaacc gtggatgatg tggtctctac aggatctgac attattattg ttggaagagg
5281 actatttgca aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg
5341 ggaagcatat ttgagaagat gcggccagca aaactaactc gagctaaatt gtaagcgtta
5401 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
5461 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
5521 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
5581 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
5641 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
5701 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
5761 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
5821 atgcgccgct acagggcgcg tc
//
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