pHIS2

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  • Model: PVT4037
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pHIS2

PVT4037   2ug                                                                   pHIS2  Data Sheet


pHIS2 Plasmid information

Promoters: TRP1, T7, T3
Replicon: pUC ori, FI ori
Plasmid classification: yeast series, yeast one hybrid vector
Plasmid size: 7207bp
Prokaryotic resistance: Kan
Selection marker: TRP1
Clone strain: DH5 alpha
Culture conditions: 37 LB, aerobic
Host cells: yeast cells
Primers for 5'sequencing: M13F:TGTAAAACGACGGCCAGT
Primers for 3'sequencing: primers were designed according to the sequence
Use: Yeast expression
 

pHIS2 Plasmid description
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.
pHIS2-plasmid


pHIS2 Plasmid Sequence
 

LOCUS       Exported                7207 bp ds-DNA     circular SYN 23-Feb-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 2016-2-23 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

//

 

Caution:
Product is for research use only!

 

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