| 质粒名称: | pH3U3-zif268 (omega) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 18046 |
| 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: | 5807bp |
| 完整质粒大小: | 5828bp |
| 载体类型: | 细菌表达 |
| 筛选标记: | URA3, HIS3 |
| 载体抗性: | 氨苄霉素 |
| 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: | zif268 binding site |
| 插入大小: | 15bp |
| 基因突变: | The preferred binding site for zif268 is inserted 10bp upstream of the -35box. This reporter plasmid is to be used as a positive test with pB1H2w2-zif268. |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Not1 (not destroyed) |
| 3’克隆位点: | EcoRI (not destroyed) |
| 5’测序引物 | HU100 |
|
文献引用方法 |
|
| 材料和方法部分: | pH3U3-zif268 (omega) was a gift from Scot Wolfe (Addgene plasmid # 18046 ; http://n2t.net/addgene:18046 ; RRID:Addgene_18046) |
| 参考文献部分: |
A systematic characterization of factors that regulate Drosophila segmentation via a bacterial one-hybrid system. Noyes MB, Meng X, Wakabayashi A, Sinha S, Brodsky MH, Wolfe SA. Nucleic Acids Res. 2008 Mar 10. ():. 10.1093/nar/gkn048 PubMed 18332042
|
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pH3U3-zif268 (omega)载体质粒基本信息
pH3U3-zif268 (omega)质粒图谱载体图谱和pH3U3-zif268 (omega)载体序列质粒序列多克隆位点信息
质粒图谱.png)
pH3U3-zif268 (omega)质粒载体简介
pH3U3-zif268 (omega)质粒序列载体序列
LOCUS pH3U3-zif268 (omega) 5828 bp DNA circular SYN 26-AUG-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5828)
AUTHORS Noyes MB, Meng X, Wakabayashi A, Sinha S, Brodsky MH, Wolfe SA
TITLE A systematic characterization of factors that regulate Drosophila
segmentation via a bacterial one-hybrid system.
JOURNAL Nucleic Acids Res. 2008 Mar 10. ():.
PUBMED 18332042
REFERENCE 2 (bases 1 to 5828)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 26, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res. 2008 Mar 10. ():."
FEATURES Location/Qualifiers
source 1..5828
/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 3779..3809
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind 3819..3841
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3838..3854
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 3850..4512
/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 4560..5363
/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(5372..5827)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5509..5530)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5721..5740
/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 cgtgggcggg acgaattctt
3781 tacactttat gcttccggct cgtatgttgt gtcgaccgag cggataacaa tttcacacag
3841 gaaacagcta tgacagagca gaaagcccta gtaaagcgta ttacaaatga aaccaagatt
3901 cagattgcga tctctttaaa gggtggtccc ctagcgatag agcactcgat cttcccagaa
3961 aaagaggcag aagcagtagc agaacaggcc acacaatcgc aagtgattaa cgtccacaca
4021 ggtatagggt ttctggacca tatgatacat gctctggcca agcattccgg ctggtcgcta
4081 atcgttgagt gcattggtga cttacacata gacgaccatc acaccactga agactgcggg
4141 attgctctcg gtcaagcttt taaagaggcc ctaggggccg tgcgtggagt aaaaaggttt
4201 ggatcaggat ttgcgccttt ggatgaggca ctttccagag cggtggtaga tctttcgaac
4261 aggccgtacg cagttgtcga acttggtttg caaagggaga aagtaggaga tctctcttgc
4321 gagatgatcc cgcattttct tgaaagcttt gcagaggcta gcagaattac cctccacgtt
4381 gattgtctgc gaggcaagaa tgatcatcac cgtagtgaga gtgcgttcaa ggctcttgcg
4441 gttgccataa gagaagccac ctcgcccaat ggtaccaacg atgttccctc caccaaaggt
4501 gttcttatgt agtgacaccg attatttaaa gctgcaggga tcccacacag gaaacagcta
4561 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc
4621 tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca
4681 ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac
4741 atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat
4801 ccgccaagta caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag
4861 tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg
4921 cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggcg gaagaagtaa
4981 caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctagcta
5041 ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg
5101 gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga
5161 cacccggtgt gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg
5221 atgatgtggt ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg
5281 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga
5341 gaagatgcgg ccagcaaaac taactcgagc taaattgtaa gcgttaatat tttgttaaaa
5401 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
5461 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
5521 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
5581 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
5641 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
5701 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
5761 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
5821 ggcgcgtc
//
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