基本信息
| 原核抗性: | 氨苄青霉素Amp |
| 筛选标记: | HIS3 |
| 克隆菌株: | 大肠杆菌DH5α |
| 培养条件: | 37℃ |
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
质粒序列
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
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
温馨提示：不可用于临床ZL。
