• 规格：20μl质粒
• 原核抗性：氨苄青霉素Amp
• 筛选标记：HIS3
• 克隆菌株：大肠杆菌DH5α
• 质粒简介：            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.
• 质粒图谱1： png/1-1G203103H1217.png

质粒序列： 点击查看序列

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 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