pRS316
pRS316
pRS316
编号 | 载体名称 |
北京华越洋生物VECT2210 | pRS316 |
pRS316载体基本信息
载体名称: | pESC-URA, pESC URA |
---|---|
Gene accession NO. : | AF063585 |
质粒类型: | 酿酒酵母蛋白表达载体 |
表达水平: | 高拷贝 |
诱导方法: | 半乳糖 |
启动子: | T7 |
克隆方法: | 多克隆位点,限制性内切酶 |
载体大小: | 4887 bp |
5' 测序引物及序列: | T7: TAATACGACTCACTATAGGG |
3' 测序引物及序列: | M13-R: CAGGAAACAGCTATGAC |
载体标签: | -- |
载体抗性: | 氨苄 |
筛选标记: | URA3 |
备注: | 利用半乳糖诱导,可以同时使两个基因在酿酒酵母中表达。 |
稳定性: | 稳定 Stable |
组成型/诱导型: | 诱导型 |
病毒/非病毒: | 非病毒 |
pRS316载体质粒图谱和多克隆位点信息
pRS316载体序列
LOCUS pRS316 4887 bp ds-DNA circular SYN 09-九月-2012 DEFINITION Yeast centromere vector with URA3 marker. ACCESSION U03442 VERSION . KEYWORDS pRS316 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4887) AUTHORS Sikorski RS, Hieter P. TITLE A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. JOURNAL Genetics 1989;122:19-27. PUBMED 2659436 REFERENCE 2 (bases 1 to 4887) AUTHORS . TITLE Direct Submission JOURNAL Exported 2013年8月13日 from SnapGene Viewer 1.5.3 http://www.snapgene.com COMMENT FEATURES Location/Qualifiers source 1..4887 /organism="synthetic DNA construct" /lab_host="Saccharomyces cerevisiae" /mol_type="other DNA" promoter 190..410 /gene="S. cerevisiae URA3" /note="URA3 promoter" /note="color: #ffffff; direction: RIGHT" CDS 411..1214 /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /note="URA3" /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /note="color: #ff7f50" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" rep_origin complement(1343..1798) /direction=LEFT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /note="color: #ffff00" CDS complement(1581..2156) /codon_start=1 /gene="lacZ?fragment" /product="fragment of?beta-galactosidase" /note="lacZ'" /note="color: #993366" /translation="MTMITPSSELTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV LERPPPRWSSNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG FPRQALNRGLPLGFRFSALRHLDPKKLD" primer_bind 1943..1959 /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: RIGHT" promoter 1966..1984 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff; direction: RIGHT" misc_feature 1993..2100 /note="MCS" /note="pBluescript multiple cloning site" /note="color: #99ccff" promoter complement(2113..2131) /note="T3 promoter" /note="promoter for bacteriophage T3 RNA polymerase" /note="color: #ffffff; direction: LEFT" primer_bind complement(2152..2168) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: LEFT" protein_bind 2176..2192 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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)." /note="color: #31849b" promoter complement(2200..2230) /note="lac promoter" /note="promoter for the E. coli lac operon" /note="This reverse directional feature has 3 segments: ???1:?2200?..?2206?/?#ffffff?/?-10 ???2:?2207?..?2224?/?#ffffff ???3:?2225?..?2230?/?#ffffff?/?-35" rep_origin complement(2554..3142) /direction=LEFT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(3313..4173) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: ???1:?3313?..?4104?/?#ccffcc ???2:?4105?..?4173?/?#ccffcc?/?signal?sequence Cleavage site after base 4104" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(4174..4278) /gene="bla" /note="AmpR promoter" /note="color: #ffffff; direction: LEFT" misc_feature 4315..4818 /note="CEN/ARS" /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" /note="color: #454f59" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc 241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca 301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat 361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag 421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata 481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat 541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata 601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt 661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc 721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg 781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac 841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat 901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg 961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg 1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg 1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg 1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg 1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta 1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt 1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta 1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat 1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca 1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc 1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta 1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg 1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg 1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg 1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg 1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca 1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta 1981 tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtgg atcccccggg 2041 ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg gcccggtacc 2101 cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct 2161 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat 2221 aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc 2281 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 2341 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 2401 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 2461 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 2521 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctcggcc cccctgacga 2581 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 2641 ccaggcgttc ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 2701 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 2761 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 2821 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 2881 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 2941 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 3001 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 3061 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 3121 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 3181 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 3241 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 3301 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 3361 tcgttcatcc atagttgcct gactgcccgt cgtgtagata actacgatac gggagggctt 3421 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 3481 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 3541 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 3601 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 3661 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 3721 gtgaaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 3781 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 3841 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 3901 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 3961 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 4021 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 4081 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 4141 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 4201 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 4261 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg 4321 tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa 4381 agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta 4441 gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc 4501 cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt 4561 ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata 4621 tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat 4681 tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat 4741 aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta 4801 agttacaggc aagcgatccg tcctaagaaa ccattattat catgacatta acctataaaa 4861 ataggcgtat cacgaggccc tttcgtc //