pGilda

$193.00

  • Model: PVT11258
  • 50 Units in Stock
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pGilda

PVT11258    Packing 2ug

 

pGilda Information
Function yeast Hybrid plasmid
Resistance Amp
Screen HIS3
Strain DH5alpha
Culture temperature 37degrees centigrade

 

pGilda Description

pGilda is a cloning vector used to generate fusions of a bait protein with the 202-residue LexA protein, which acts as a DNA-binding domain in the MATCHMAKER LexA Two-Hybrid System . The hybrid protein is expressed in yeast cells from the tightly regulated GAL1 promoter (PGAL1); transcription is repressed in the presence of glucose and induced in the the absence of glucose and presence of galactose . pGilda carries the HIS3 gene for selection in His– auxotrophic yeast strains and is compatible with MATCHMAKER LexA Libraries. This vector replicates autonomously in both E. coli (using the Col E1 ori) and in S. cerevisiae (using the CEN/ARS ori). Ampr is used for selection in E. coli.

 

pGilda Sequence

LOCUS       Exported                6570 bp ds-DNA     circular SYN 03-DEC-2017
DEFINITION  Centromeric yeast two-hybrid vector for galactose-regulated 
            expression of LexA fused to the N-terminus of a protein.
FEATURES             Location/Qualifiers
     source          1..6570
                     /organism="synthetic DNA construct"
                     /lab_host="Saccharomyces cerevisiae"
                     /mol_type="other DNA"
     promoter        8..672
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    224..341
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     CDS             692..1297
                     /codon_start=1
                     /gene="lexA"
                     /product="E. coli DNA-binding protein that represses genes 
                     involved in the SOS response to DNA damage"
                     /label=LexA
                     /note="The N-terminal DNA-binding domain recognizes the 
                     16-bp palindromic sequence 5'-CTGTATATATATACAG-3'."
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPLLAQQHIEGHYQVDPSL
                     FKPNADFLLRVSGMSMKDIGIMDGDLLAVHKTQDVRNGQVVVARIDDEVTVKRLKKQGN
                     KVELLPENSEFKPIVVDLRQQSFTIEGLAVGVIRNGDWL"
     misc_feature    1298..1336
                     /label=MCS
                     /note="multiple cloning site"
     terminator      1363..1550
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for alcohol dehydrogenase 
                     1"
     rep_origin      1882..2337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(2598..3257)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(3258..3445)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     misc_feature    3829..4332
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4369..4473
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4474..5334
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5505..6093
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cttgaatttt caaaaattct tacttttttt ttggatggac gcaaagaagt ttaataatca
       61 tattacatgg cattaccacc atatacatat ccatatacat atccatatct aatcttactt
      121 atatgttgtg gaaatgtaaa gagccccatt atcttagcct aaaaaaacct tctctttgga
      181 actttcagta atacgcttaa ctgctcattg ctatattgaa gtacggatta gaagccgccg
      241 agcgggtgac agccctccga aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg
      301 cgttcctgaa acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa gattctacaa
      361 tactagcttt tatggttatg aagaggaaaa attggcagta acctggcccc acaaaccttc
      421 aaatgaacga atcaaattaa caaccatagg atgataatgc gattagtttt ttagccttat
      481 ttctggggta attaatcagc gaagcgatga tttttgatct attaacagat atataaatgc
      541 aaaaactgca taaccacttt aactaatact ttcaacattt tcggtttgta ttacttctta
      601 ttcaaatgta ataaaagtat caacaaaaaa ttgttaatat acctctatac tttaacgtca
      661 aggagaaaaa accccggatc aagggtgcga tatgaaagcg ttaacggcca ggcaacaaga
      721 ggtgtttgat ctcatccgtg atcacatcag ccagacaggt atgccgccga cgcgtgcgga
      781 aatcgcgcag cgtttggggt tccgttcccc aaacgcggct gaagaacatc tgaaggcgct
      841 ggcacgcaaa ggcgttattg aaattgtttc cggcgcatca cgcgggattc gtctgttgca
      901 ggaagaggaa gaagggttgc cgctggtagg tcgtgtggct gccggtgaac cacttctggc
      961 gcaacagcat attgaaggtc attatcaggt cgatccttcc ttattcaagc cgaatgctga
     1021 tttcctgctg cgcgtcagcg ggatgtcgat gaaagatatc ggcattatgg atggtgactt
     1081 gctggcagtg cataaaactc aggatgtacg taacggtcag gtcgttgtcg cacgtattga
     1141 tgacgaagtt accgttaagc gcctgaaaaa acagggcaat aaagtcgaac tgttgccaga
     1201 aaatagcgag tttaaaccaa ttgtcgtaga tcttcgtcag cagagcttca ccattgaagg
     1261 gctggcggtt ggggttattc gcaacggcga ctggct
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