pGilda/ pGilda质粒/ pGilda Plasmid_详细资料

产品信息

货号： PVT11258 CAS号：无供应商： LSM Bio 数量：货源充足英文名： pGilda 保质期： 6month 保存条件：负20摄氏度规格： 2ug

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。

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