• 规格：20μl质粒
• 原核抗性：氨苄青霉素Amp
• 筛选标记：LEU2
• 克隆菌株：大肠杆菌DH5α
• 质粒简介：            YEplac181是一个酿酒酵母表达质粒。
• 质粒图谱1： png/1-1G221151041333.png

质粒序列： 点击查看序列

LOCUS       Exported                5741 bp ds-DNA     circular SYN 21-DEC-2017

DEFINITION  Yeast episomal plasmid with a LEU2 marker.

COMMENT     The sequence of this plasmid was reconstructed using the description

            from Gietz and Sugino.

FEATURES             Location/Qualifiers

     source          1..5741

                     /organism="synthetic DNA construct"

                     /lab_host="Saccharomyces cerevisiae"

                     /mol_type="other DNA"

     promoter        142..172

                     /label=lac promoter

                     /note="promoter for the E. coli lac operon"

     protein_bind    180..196

                     /label=lac operator

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="The lac repressor binds to the lac operator to 

                     inhibit transcription in E. coli. This inhibition can be 

                     relieved by adding lactose or 

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

     CDS             216..500

                     /codon_start=1

                     /gene="lacZ"

                     /product="LacZ-alpha fragment of beta-galactosidase"

                     /label=lacZ-alpha

                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR

                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRI"

     misc_feature    233..289

                     /label=MCS

                     /note="pUC19 multiple cloning site"

     rep_origin      745..2091

                     /label=2u ori

                     /note="yeast 2u plasmid origin of replication"

     CDS             complement(2227..3321)

                     /codon_start=1

                     /gene="S. cerevisiae LEU2"

                     /product="3-isopropylmalate dehydrogenase, required for 

                     leucine biosynthesis"

                     /label=LEU2

                     /note="yeast auxotrophic marker"

                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL

                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA

                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT

                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ

                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF

                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG

                     DLGGSNSTTEVGDAVAEEVKKILA"

     promoter        complement(3322..3729)

                     /gene="S. cerevisiae LEU2"

                     /label=LEU2 promoter

     promoter        3835..3939

                     /gene="bla"

                     /label=AmpR promoter

     CDS             3940..4800

                     /codon_start=1

                     /gene="bla"

                     /product="beta-lactamase"

                     /label=AmpR

                     /note="confers resistance to ampicillin, carbenicillin, and

                     related antibiotics"

                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI

                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS

                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW

                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA

                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS

                     LIKHW"

     rep_origin      4971..5559

                     /direction=RIGHT

                     /label=ori

                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 

                     replication"

ORIGIN

        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca

       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct

      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat

      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc

      241 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg

      301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac

      361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac

      421 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt

      481 gcggtatttc acaccgcata tatcggatcg tacttgttac ccatcattga attttgaaca

      541 tccgaacctg ggagttttcc ctgaaacaga tagtatattt gaacctgtat aataatatat

      601 agtctagcgc tttacggaag acaatgtatg tatttcggtt cctggagaaa ctattgcatc

      661 tattgcatag gtaatcttgc acgtcgcatc cccggttcat tttctgcgtt tccatcttgc

      721 acttcaatag catatctttg ttaacgaagc atctgtgctt cattttgtag aacaaaaatg

      781 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa

      841 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa

      901 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa

      961 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc

     1021 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt

     1081 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa

     1141 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac

     1201 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc

     1261 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg

     1321 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata

     1381 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct

     1441 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc

     1501 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc

     1561 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat

     1621 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcttcagag

     1681 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac

     1741 ttcggaatag gaacttcaaa gcgtttccga aaacgagcgc ttccgaaaat gcaacgcgag

     1801 ctgcgcacat acagctcact gttcacgtcg cacctatatc tgcgtgttgc ctgtatatat

     1861 atatacatga gaagaacggc atagtgcgtg tttatgctta aatgcgtact tatatgcgtc

     1921 tatttatgta ggatgaaagg tagtctagta cctcctgtga tattatccca ttccatgcgg

     1981 ggtatcgtat gcttccttca gcactaccct ttagctgttc tatatgctgc cactcctcaa

     2041 ttggattagt ctcatccttc aatgctatca tttcctttga tattggatcg atccgatgat

     2101 aagctgtcaa acatgagaat taattctacc ctatgaacat attccatttt gtaatttcgt

     2161 gtcgtttcta ttatgaattt catttataaa gtttatgtac aaatatcata aaaaaagaga

     2221 atctttttaa gcaaggattt tcttaacttc ttcggcgaca gcatcaccga cttccgtggt

     2281 actgttggaa ccacctaaat caccagttct gatacctgca tccaaaacct ttttaactgc

     2341 atcttcaatg gccttacctt cttcaggcaa gttcaatgac aatttcaaca tcattgcagc

     2401 agacaagata gtggcgatag ggttgacctt attctttggc aaatctggag cagaaccgtg

     2461 gcatggttcg tacaaaccaa atgcggtgtt cttgtctggc aaagaggcca aggacgcaga

     2521 tggcaacaaa cccaaggaac ctgggataac ggaggcttca tcggagatga tatcaccaaa

     2581 catgttgctg gtgattataa taccatttag gtgggttggg ttcttaacta ggatcatggc

     2641 ggcagaatca atcaattgat gttgaacctt caatgtagga aattcgttct tgatggtttc

     2701 ctccacagtt tttctccata atcttgaaga ggccaaaaca ttagctttat ccaaggacca

     2761 aataggcaat ggtggctcat gttgtagggc catgaaagcg gccattcttg tgattctttg

     2821 cacttctgga acggtgtatt gttcactatc ccaagcgaca ccatcaccat cgtcttcctt

     2881 tctcttacca aagtaaatac ctcccactaa ttctctgaca acaacgaagt cagtaccttt

     2941 agcaaattgt ggcttgattg gagataagtc taaaagagag tcggatgcaa agttacatgg

     3001 tcttaagttg gcgtacaatt gaagttcttt acggattttt agtaaacctt gttcaggtct

     3061 aacactacct gtaccccatt taggaccacc cacagcacct aacaaaacgg catcaacctt

     3121 cttggaggct tccagcgcct catctggaag tgggacacct gtagcatcga tagcagcacc

     3181 accaattaaa tgattttcga aatcgaactt gacattggaa cgaacatcag aaatagcttt

     3241 aagaacctta atggcttcgg ctgtgatttc ttgaccaacg tggtcacctg gcaaaacgac

     3301 gatcttctta ggggcagaca ttagaatggt atatccttga aatatatata tatatattgc

     3361 tgaaatgtaa aaggtaagaa aagttagaaa gtaagacgat tgctaaccac ctattggaaa

     3421 aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc

     3481 atgaacgctt ctctattcta tatgaaaagc cggttccgcg gctctcacct ttcctttttc

     3541 tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat taacaaaaaa

     3601 tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc gttatgttga

     3661 ggaaaaaaat aatggttgct aagagattcg aactcttgca tcttacgata cctgagtatt

     3721 cccacagtta attcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat

     3781 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga

     3841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa

     3901 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt

     3961 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg

     4021 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg

     4081 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg

     4141 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag

     4201 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca

     4261 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg

     4321 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc

     4381 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg

     4441 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg

     4501 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac

     4561 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg

     4621 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg

     4681 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact

     4741 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa

     4801 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt

     4861 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag

     4921 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct