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 <
温馨提示：不可用于临床ZL。
