| 质粒名称: | pCEV-G4-Km |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 46819 |
| 存储实验室: | Lars Nielsen, Claudia Vickers |
|
载体骨架基本信息 |
|
| 载体名称: | pCEV-G2-Km |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | PGK1启动子替换为HXT7启动子。 |
| 载体类型: | 酵母表达 |
| 筛选标记: | G418 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | TEF1 and HXT7 |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCEV-G4-Km was a gift from Lars Nielsen & Claudia Vickers (Addgene plasmid # 46819 ; http://n2t.net/addgene:46819 ; RRID:Addgene_46819) |
| 参考文献部分: | Dual gene expression cassette vectors with antibiotic selection markers for engineering in Saccharomyces cerevisiae. Vickers CE, Bydder SF, Zhou Y, Nielsen LK. Microb Cell Fact. 2013 Oct 25;12(1):96. 10.1186/1475-2859-12-96 PubMed 24161108 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pCEV-G4-Km 6574 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION TEF1 and HXT7 promoter controlled expression cassettes with G418 resistance for yeast transformation. ACCESSION . VERSION . KEYWORDS pCEV-G4-Km SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6574) AUTHORS Vickers CE, Bydder SF, Zhou Y, Nielsen LK TITLE Dual gene expression cassette vectors with antibiotic selection markers for engineering in Saccharomyces cerevisiae. JOURNAL Microb Cell Fact. 2013 Oct 25;12(1):96. PUBMED 24161108 REFERENCE 2 (bases 1 to 6574) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6574 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 2..167 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind 88..112 /label=pHybLex-R /note="ADH terminator, reverse primer" CDS complement(315..338) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter 947..1353 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" primer_bind 1373..1392 /label=T7 /note="T7 promoter, forward primer" promoter 1373..1391 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1409..1438 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" terminator 1468..1657 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(1729..1746) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1900..2488) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1980..1999) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2659..3519) /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" primer_bind 3282..3301 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3520..3624) /gene="bla" /label=AmpR promoter rep_origin 3651..4993 /label=2u ori /note="yeast 2u plasmid origin of replication" protein_bind complement(4614..4661) /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." protein_bind complement(5013..5046) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." gene 5101..6457 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 5101..5444 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 5445..6254 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(5512..5531) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" terminator 6260..6457 /label=TEF terminator /note="Ashbya gossypii TEF terminator" protein_bind complement(6520..6553) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." ORIGIN 1 ggagcgacct catgctatac ctgagaaagc aacctgacct acaggaaaga gttactcaag 61 aataagaatt ttcgttttaa aacctaagag tcactttaaa atttgtatac acttattttt 121 tttataactt atttaataat aaaaatcata aatcataaga aattcgctta tttagaagtg 181 tcaacaacgt atctaccaac gatttgaccc ttttccatct tttcgtaaat ttctggcaag 241 gtagacaagc cgacaacctt gattggagac ttgaccaaac ctctggcgaa gaattgttaa 301 ttaagagctc agatcttatc gtcgtcatcc ttgtaatcca tcgatactag ttttttgatt 361 aaaattaaaa aaactttttg tttttgtgtt tattctttgt tcttagaaaa gacaagttga 421 gcttgtttgt tcttgatgtt ttattatttt acaatagctg caaatgaaga atagattcga 481 acattgtgaa gtattggcat atatcgtctc tatttatact tttttttttt cagttctagt 541 atattttgta ttttcctcct tttcattctt tcagttgcca ataagttaca ggggatctcg 601 aaagatggtg gggatttttc cttgaaagac gactttttgc catctaattt ttccttgttg 661 cctctgaaaa ttatccagca gaagcaaatg taaaagatga acctcagaag aacacgcagg 721 ggcccgaaat tgttcctacg agaagtagtg ggtcataaaa agtttattcc ctggaaaaaa 781 aattttgcgt tgcctttctg gagaattttt tcgaattagc gtgctgccac tgcatgcatt 841 tctgagaagt gtgggcattc ttccaccagt tgttcctcct aaaaaaaaaa agatttccta 901 ccccgcacaa attcctgcat acccctcatt tccacggggc cggccgcaca caccatagct 961 tcaaaatgtt tctactcctt ttttactctt ccagattttc tcggactccg cgcatcgccg 1021 taccacttca aaacacccaa gcacagcata ctaaatttcc cctctttctt cctctagggt 1081 gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa gagaccgcct cgtttctttt 1141 tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc tttttcttga aaattttttt 1201 ttttgatttt tttctctttc gatgacctcc cattgatatt taagttaata aacggtcttc 1261 aatttctcaa gtttcagttt catttttctt gttctattac aacttttttt acttcttgct 1321 cattagaaag aaagcatagc aatctaatct aagttttaat tacaaggatc cgtaatacga 1381 ctcactatag ggcccgggcg tcgacatgga acagaagttg atttccgaag aagacctcga 1441 gtaagcttgg taccgcggct agctaagatc cgctctaacc gaaaaggaag gagttagaca 1501 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt 1561 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact 1621 gaaaaccttg cttgagaagg ttttgggacg ctcgaagatc cagctgcatt aatgaatcgg 1681 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 1741 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 1801 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 1861 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 1921 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 1981 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 2041 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 2101 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 2161 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 2221 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 2281 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 2341 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 2401 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 2461 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 2521 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 2581 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 2641 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 2701 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 2761 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 2821 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 2881 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 2941 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 3001 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 3061 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 3121 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 3181 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 3241 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 3301 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 3361 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 3421 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 3481 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 3541 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 3601 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aacgaagcat 3661 ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga 3721 atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa 3781 gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac 3841 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca 3901 acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc 3961 taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg 4021 ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt 4081 ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc 4141 gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg 4201 catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga 4261 acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt 4321 gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact 4381 agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt 4441 ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag 4501 caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt 4561 ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc tctgaagttc 4621 ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt ccgaaaacga 4681 gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac gtcgcaccta 4741 tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg cgtgtttatg 4801 cttaaatgcg tacttatatg cgtctattta tgtaggatga aaggtagtct agtacctcct 4861 gtgatattat cccattccat gcggggtatc gtatgcttcc ttcagcacta ccctttagct 4921 gttctatatg ctgccactcc tcaattggat tagtctcatc cttcaatgct atcatttcct 4981 ttgatattgg atcatggtag acaaccctta atataacttc gtataatgta tgctatacga 5041 agttattagg tctagagatc tgtttagctt gcctcgtccc cgccgggtca cccggccagc 5101 gacatggagg cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga 5161 tgtgactgtc gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata 5221 cattttgatg gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag 5281 cagggaaacg ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga 5341 aatataaaag gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc 5401 ttgctaggat acagttctca catcacatcc gaacataaac aaccatgggt aaggaaaaga 5461 ctcacgtttc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat 5521 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg 5581 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg 5641 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta 5701 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggcaaa acagcattcc 5761 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc 5821 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc 5881 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg 5941 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataag cttttgccat 6001 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg 6061 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg 6121 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt 6181 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg 6241 atgagttttt ctaatcagta ctgacaataa aaagattctt gttttcaaga acttgtcatt 6301 tgtatagttt ttttatattg tagttgttct attttaatca aatgttagcg tgatttatat 6361 tttttttcgc ctcgacatca tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc 6421 atgcgtcaat cgtatgtgaa tgctggtcgc tatactgctg tcgattcgat actaacgccg 6481 ccatccagtg tcgaaaacga gctctcgaga acccttaata taacttcgta taatgtatgc 6541 tatacgaagt tattaggtga tatcagatcc acta //
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：pSK+KanaRpsL
- 下一篇：pHIV-Luciferase
