pGilda

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  • Model: PVT11258
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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.

 

Publication for PVT11258 pGilda

ARTICLE:A Tale of Two-Hybrids: Investigating the Interactomes of Cullin-Associated Proteins

Author:Reitter, Elyse.

Portland State University ProQuest Dissertations Publishing,  2022. 28967284.

 

pGilda Sequence

LOCUS       Exported                6570 bp ds-DNA    circular SYN 25-2-2014
DEFINITION  Centromeric yeast two-hybrid vector for galactose-regulated 
            expression of LexA fused to the N-terminus of a protein.
ACCESSION   .
VERSION     .
KEYWORDS    pGilda
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6570)
  AUTHORS   Clontech
  TITLE     Direct Submission
  JOURNAL   Exported 2017/3/1 from SnapGene 2.3.2
            http://www.snapgene.com
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"
                     /note="GAL1,10 promoter"
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    224..341
                     /bound_moiety="Gal4"
                     /note="UAS"
                     /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"
                     /note="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
                     /note="MCS"
                     /note="multiple cloning site"
     terminator      1363..1550
                     /gene="S. cerevisiae ADH1"
                     /note="ADH1 terminator"
                     /note="transcription terminator for alcohol dehydrogenase 
                     1"
     primer_bind     complement(1721..1737)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1882..2337
                     /direction=RIGHT
                     /note="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"
                     /note="HIS3"
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(3258..3445)
                     /gene="S. cerevisiae HIS3"
                     /note="HIS3 promoter"
     misc_feature    3829..4332
                     /note="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4369..4473
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             4474..5334
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5505..6093
                     /direction=RIGHT
                     /note="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 ctggctggaa ttcccgggga tccgtcgacc
     1321 atggcggccg ctcgagtcga cctgcagcca agctaattcc gggcgaattt cttatgattt
     1381 atgattttta ttattaaata agttataaaa aaaataagtg tatacaaatt ttaaagtgac
     1441 tcttaggttt taaaacgaaa attcttgttc ttgagtaact ctttcctgta ggtcaggttg
     1501 ctttctcagg tatagcatga ggtcgctctt attgaccaca cctctaccgg catgccgagc
     1561 aaatgcctgc aaatcgctcc ccatttcacc caattgtaga tatgctaact ccagcaatga
     1621 gttgatgaat ctcggtgtgt attttatgtc ctcagaggac aacacctgtt gtaatccgtc
     1681 cgagctccaa ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac
     1741 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1801 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     1861 gcctgaatgg cgaatggcgc gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1921 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1981 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2041 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2101 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     2161 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     2221 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     2281 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcct
     2341 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat gatccgtcga
     2401 gttcaagaga aaaaaaaaga aaaagcaaaa agaaaaaagg aaagcgcgcc tcgttcagaa
     2461 tgacacgtat agaatgatgc attaccttgt catcttcagt atcatactgt tcgtatacat
     2521 acttactgac attcataggt atacatatat acacatgtat atatatcgta tgctgcagct
     2581 ttaaataatc ggtgtcacta cataagaaca cctttggtgg agggaacatc gttggtacca
     2641 ttgggcgagg tggcttctct tatggcaacc gcaagagcct tgaacgcact ctcactacgg
     2701 tgatgatcat tcttgcctcg cagacaatca acgtggaggg taattctgct agcctctgca
     2761 aagctttcaa gaaaatgcgg gatcatctcg caagagagat ctcctacttt ctccctttgc
     2821 aaaccaagtt cgacaactgc gtacggcctg ttcgaaagat ctaccaccgc tctggaaagt
     2881 gcctcatcca aaggcgcaaa tcctgatcca aaccttttta ctccacgcgc cagtagggcc
     2941 tctttaaaag cttgaccgag agcaatcccg cagtcttcag tggtgtgatg gtcgtctatg
     3001 tgtaagtcac caatgcactc aacgattagc gaccagccgg aatgcttggc cagagcatgt
     3061 atcatatggt ccagaaaccc tatacctgtg tggacgttaa tcacttgcga ttgtgtggcc
     3121 tgttctgcta ctgcttctgc ctctttttct gggaagatcg agtgctctat cgctagggga
     3181 ccacccttta aagagatcgc aatctgaatc ttggtttcat ttgtaatacg ctttactagg
     3241 gctttctgct ctgtcatctt tgccttcgtt tatcttgcct gctcattttt tagtatattc
     3301 ttcgaagaaa tcacattact ttatataatg tataattcat tatgtgataa tgccaatcgc
     3361 taagaaaaaa aaagagtcat ccgctaggtg gaaaaaaaaa aatgaaaatc attaccgagg
     3421 cataaaaaaa tatagagtgt actagaggag gccaagagta atagaaaaag aaaattgcgg
     3481 gaaaggactg tgttatgact tccctgacta atgccgtgtt caaacgatac ctggcagtga
     3541 ctcctagcgc tcaccaagct cttaaaacgg aattatggtg cactctcagt acaatctgct
     3601 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     3661 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3721 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     3781 gcctattttt ataggttaat gtcatgataa taatggtttc ttaggacgga tcgcttgcct
     3841 gtaacttaca cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt
     3901 ttatttattt ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt
     3961 tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt
     4021 tacatatata tttattagac aagaaaagca gattaaatag atatacattc gattaacgat
     4081 aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac
     4141 aattcggcat taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga
     4201 tccccctaga gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt
     4261 tttactttct atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt
     4321 atagcacgtg atgaaaagga cccaggtggc acttttcggg gaaatgtgcg cggaacccct
     4381 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4441 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4501 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4561 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4621 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4681 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4741 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     4801 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     4861 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     4921 tttcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     4981 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5041 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5101 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5161 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5221 gatggtaagc cctcccgtat cgtagttatc tacacgacgg gcagtcaggc aactatggat
     5281 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5341 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5401 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5461 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5521 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5581 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5641 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5701 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5761 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5821 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5881 tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5941 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc aggggggaac
     6001 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6061 tgatgctcgt caggggggcc gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6121 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6181 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6241 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6301 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6361 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     6421 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     6481 ggaaacagct atgaccatga ttaccccaag ctcgaaatta accctcacta aagggaacaa
     6541 aagctggtac cgggcccccc ctcgaaattc
//

Caution:
1.  This product is FOR RESEARCH USE ONLY!
2.  The item is lyophilized form, Please take the powder plasmid by centrifugation at 5000rpm/min for 1min. Add 20μl ddH2O in to the tube of plasmid.
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