pHIS2酵母单杂交质粒

pHIS2酵母单杂交质粒

价格: ¥1000

品牌:钦诚生物

货号:QC0421

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保存条件 :负20度

保质期 :2年

供应商 :钦诚生物

规格 :5ug

基本信息

启动子: TRP1 ,T7,T3
复制子: pUC ori,FI ori
质粒分类: 酵母系列,酵母单杂交载体
质粒大小: 7207bp
原核抗性: Kan
筛选标记: TRP1
克隆菌株: DH5α
培养条件: 37℃,有氧 LB
表达宿主: 酵母细胞
5'测序引物: M13F:TGTAAAACGACGGCCAGT
3'测序引物: 根据序列设计引物

质粒简介


    pHIS2 is a reporter vector that can be used in yeast one-hybrid assays to identify and characterizeDNA-binding proteins. The vector was specifically designed for use with the BD Matchmaker™ One-Hybrid Library Construction & Screening Kit (#K1617-1). It contains a HIS3 nutritional reporter gene,located downstream of a multiple cloning site (MCS) and the minimal promoter of the HIS3 locus(PminHIS3). Cis-acting DNA sequences, or DNA target elements, can be inserted into the MCS andused as baits to screen GAL4 AD/cDNA fusion libraries for proteins that interact with the targetsequence. A protein-DNA (or one-hybrid) interaction can be detected by performing the assay in ayeast strain such as Y187 that is auxotrophic for histidine. Positive one-hybrid interactions driveexpression of the HIS3 reporter gene, which enables the host cell to grow on histidine-deficientmedia.

质粒图谱


质粒序列


LOCUS       Exported                7207 bp ds-DNA     circular SYN 23-二月-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7207)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2016年2月23日 from SnapGene Viewer 3.0.3
FEATURES             Location/Qualifiers
     source          1..7207
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             152..814
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /note="HIS3"
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        2576..2857
                     /gene="S. cerevisiae TRP1"
                     /note="TRP1 promoter"
     CDS             2858..3532
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /note="TRP1"
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3614..3632)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3653..3669)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3677..3693
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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(3701..3731)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3746..3767
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4055..4643)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4814..5608)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHDDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5609..5713)
                     /gene="bla"
                     /note="AmpR promoter"
     misc_feature    5746..6247
                     /note="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     rep_origin      complement(6564..7019)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     7164..7180
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7187..7205
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gaattcccgg ggagctcacg cgttcgcgaa tcgatccgcg gtctagaaat tcctggcatt
       61 atcacataat gaattataca ttatataaag taatgtgatt tcttcgaaga atatactaaa
      121 aaatgagcag gcaagataaa cgaaggcaaa gatgacagag cagaaagccc tagtaaagcg
      181 tattacaaat gaaaccaaga ttcagattgc gatctcttta aagggtggtc ccctagcgat
      241 agagcactcg atcttcccag aaaaagaggc agaagcagta gcagaacagg ccacacaatc
      301 gcaagtgatt aacgtccaca caggtatagg gtttctggac catatgatac atgctctggc
      361 caagcattcc ggctggtcgc taatcgttga gtgcattggt gacttacaca tagacgacca
      421 tcacaccact gaagactgcg ggattgctct cggtcaagct tttaaagagg ccctaggggc
      481 cgtgcgtgga gtaaaaaggt ttggatcagg atttgcgcct ttggatgagg cactttccag
      541 agcggtggta gatctttcga acaggccgta cgcagttgtc gaacttggtt tgcaaaggga
      601 gaaagtagga gatctctctt gcgagatgat cccgcatttt cttgaaagct ttgcagaggc
      661 tagcagaatt accctccacg ttgattgtct gcgaggcaag aatgatcatc accgtagtga
      721 gagtgcgttc aaggctcttg cggttgccat aagagaagcc acctcgccca atggtaccaa
      781 cgatgttccc tccaccaaag gtgttcttat gtagtgacac cgattattta aagctgcagc
      841 atacgatata tatacatgtg tatatatgta tacctatgaa tgtcagtaag tatgtatacg
      901 aacagtatga tactgaagat gacaaggtaa tgcatcattc tatacgtgtc attctgaacg
      961 aggcgcgctt tccttttttc tttttgcttt ttcttttttt ttctcttgaa ctcgagaaaa
     1021 aaaatataaa agagatggag gaacgggaaa aagttagttg tggtgatagg tggcaagtgg
     1081 tattccgtaa gaacaacaag aaaagcattt catattatgg ctgaactgag cgaacaagtg
     1141 caaaatttaa gcatcaacga caacaacgag aatggttatg ttcctcctca cttaagagga
     1201 aaaccaagaa gtgccagaaa taacatgagc aactacaata acaacaacgg cggctacaac
     1261 ggtggccgtg gcggtggcag cttctttagc aacaaccgtc gtggtggtta cggcaacggt
     1321 ggtttcttcg gtggaaacaa cggtggcagc agatctaacg gccgttctgg tggtagatgg
     1381 atcgatggca aacatgtccc agctccaaga aacgaaaagg ccgagatcgc catatttggt
     1441 gtccccgagg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     1501 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     1561 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     1621 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     1681 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgac cgatgccctt
     1741 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     1801 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     1861 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     1921 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     1981 cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct
     2041 ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg
     2101 catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg
     2161 acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcattg gaccgctgat
     2221 cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg
     2281 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     2341 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga
     2401 gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc
     2461 atcgcgtccg ccatctccag cagccgcacg cggcgcatcg gggggggggt ttcaattcaa
     2521 ttcatcattt tttttttatt cttttttttg atttcggttt ctttgaaatt tttttaacga
     2581 cattactata tatataatat aggaagcatt taatagaaca gcatcgtaat atatgtgtac
     2641 tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa aaatagcttg
     2701 tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa gaattaattc
     2761 ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat tgagcacgtg
     2821 agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta atttcacagg
     2881 tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg cagaatgtgc
     2941 tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa agagaacaat
     3001 tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa atagttcagg
     3061 cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg ttttggctct
     3121 ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt ggcaagaata
     3181 ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag actgcaacat
     3241 actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg attcagaagc
     3301 aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg gaaggcaaga
     3361 gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa atgttggtga
     3421 tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg
     3481 tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat aggttattac
     3541 tgagtagtat ttatttaagt attgtttgtg cacttgccct agagtcgacc tgcaggcatg
     3601 caagcttttg ttccctttag tgagggttaa tttcgagctt ggcgtaatca tggtcatagc
     3661 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3721 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3781 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3841 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3901 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4621 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4741 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4801 cttggtctga cagtcagaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
     4861 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
     4921 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
     4981 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     5041 cgtcatgggt cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca
     5101 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     5161 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     5221 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     5281 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     5341 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     5401 gccacgatag ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct
     5461 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     5521 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     5581 ctgcgtgcaa tccatcttgt tcaatcatac tcttcctttt tcaatattat tgaagcattt
     5641 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5701 taggggttcc gcgcacattt ccccgaaaag tgccacctgc ggacggatcg cttgcctgta
     5761 acttacacgc gcctcgtatc ttttaatgat ggaataattt gggaatttac tctgtgttta
     5821 tttattttta tgttttgtat ttggatttta gaaagtaaat aaagagtaga agagttacgg
     5881 aatgaagaaa aaaaaataaa caaaggttta aaaaatttca acaaaaagcg tactttacat
     5941 atatatttat tagacaagaa aagcagatta aatagatata cattcgatta acgataagta
     6001 aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc
     6061 ggcattaata cctgagagca ggaagagcaa gataaaaggn agtatttgtt ggcgatcccc
     6121 ctagagtctt ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac
     6181 tttctatttt taatttatat atttatatta aaaaatttaa attataatta tttttatagc
     6241 acgtgatgaa aaggaccgac gtctaagaaa ccattattat catgacatta acctataaaa
     6301 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6361 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6421 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     6481 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     6541 taaggagaaa ataccgcatc aggaaattgt aaacgttaat attttgttaa aattcgcgtt
     6601 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     6661 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     6721 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     6781 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6841 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6901 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6961 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     7021 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     7081 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     7141 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact
     7201 atagggc

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