| 质粒名称: | pESC-leu2d(empty) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 20120 |
| 存储实验室: | Jay Keasling |
|
载体骨架基本信息 |
|
| 载体名称: | pESC-leu2 |
| 载体出品公司: | stratagene |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 酵母表达，合成生物学 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 未知 |
| 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 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | n/a |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pESC-leu2d(empty) was a gift from Jay Keasling (Addgene plasmid # 20120 ; http://n2t.net/addgene:20120 ; RRID:Addgene_20120) |
| 参考文献部分: | Production of the antimalarial drug precursor artemisinic acid in engineered yeast. Ro DK, Paradise EM, Ouellet M, Fisher KJ, Newman KL, Ndungu JM, Ho KA, Eachus RA, Ham TS, Kirby J, Chang MC, Withers ST, Shiba Y, Sarpong R, Keasling JD. Nature. 2006 Apr 13. 440(7086):940-3. 10.1038/nature04640 PubMed 16612385 |
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
1质粒图谱.png)
载体简介
载体序列
LOCUS pESC-leu2d(empty) 7346 bp ds-DNA circular SYN 08-JAN-2020
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pESC-leu2d(empty)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7346)
AUTHORS Ro DK, Paradise EM, Ouellet M, Fisher KJ, Newman KL, Ndungu JM, Ho
KA, Eachus RA, Ham TS, Kirby J, Chang MC, Withers ST, Shiba Y,
Sarpong R, Keasling JD
TITLE Production of the antimalarial drug precursor artemisinic acid in
engineered yeast.
JOURNAL Nature. 2006 Apr 13. 440(7086):940-3.
PUBMED 16612385
REFERENCE 2 (bases 1 to 7346)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 8, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7346
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(29..51)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 151..170
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
CDS complement(664..1758)
/codon_start=1
/gene="S. cerevisiae LEU2"
/product="3-isopropylmalate dehydrogenase, required for
leucine biosynthesis"
/label=LEU2
/note="yeast auxotrophic marker"
/translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
DLGGSNSTTEVGDAVAEEVKKILA"
rep_origin complement(2066..2521)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2203..2224)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2415..2434
/label=F1ori-R
/note="F1 origin, reverse primer"
terminator 2600..2765
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind 2686..2710
/label=pHybLex-R
/note="ADH terminator, reverse primer"
CDS complement(2913..2936)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
promoter 2983..3647
/gene="GAL1 and GAL10 genes of S. cerevisiae"
/label=GAL1,10 promoter
/note="divergent inducible promoter, regulated by Gal4"
protein_bind 3199..3316
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 3611..3634
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
primer_bind 3658..3677
/label=T7
/note="T7 promoter, forward primer"
promoter 3658..3676
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 3694..3723
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
terminator 3753..3942
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(4014..4031)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4185..4773)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4265..4284)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4944..5804)
/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 5567..5586
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5805..5909)
/gene="bla"
/label=AmpR promoter
rep_origin 5936..7278
/label=2u ori
/note="yeast 2u plasmid origin of replication"
protein_bind complement(6899..6946)
/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)."
primer_bind 7319..7337
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
181 accatatcga ctacgtcgtt aaggccgttt ctgacagagt aaaattcttg agggaacttt
241 caccattatg ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc
301 attgagtgtt ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa
361 attaggaatc atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct
421 ccttgtcaat attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta
481 tcaatttgct tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg
541 tagattgcgt atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc
601 gtttctatta tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc
661 tttttaagca aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact
721 gttggaacca cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc
781 ttcaatggcc ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga
841 caagatagtg gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca
901 tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg
961 caacaaaccc aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat
1021 gttgctggtg attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc
1081 agaatcaatc aattgatgtt gaaccttcaa tgtagggaat tcgttcttga tggtttcctc
1141 cacagttttt ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat
1201 aggcaatggt ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac
1261 ttctggaacg gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct
1321 cttaccaaag taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc
1381 aaattgtggc ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct
1441 taagttggcg tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac
1501 actaccggta ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt
1561 ggaggcttcc agcgcctcat ctggaagtgg aacacctgta gcatcgatag cagcaccacc
1621 aattaaatga ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag
1681 aaccttaatg gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat
1741 cttcttaggg gcagacatta caatggtata tccttgaaat atatataaaa aaaggcgcct
1801 tagaccgctc ggccaaacaa ccaattactt gttgagaaat agagtataat tatcctataa
1861 atataacgtt tttgaacaca catgaacaag gaagtacagg acaattgatt ttgaagagaa
1921 tgtggatttt gatgtaattg ttgggattcc atttttaata aggcaataat attaggtatg
1981 tagatatact agaagttctc ctcgaccggt cgatatgcgg tgtgaaatac cgcacagatg
2041 cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg
2101 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
2161 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
2221 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
2281 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
2341 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
2401 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
2461 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
2521 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
2581 tattacgcca gctgaattgg agcgacctca tgctatacct gagaaagcaa cctgacctac
2641 aggaaagagt tactcaagaa taagaatttt cgttttaaaa cctaagagtc actttaaaat
2701 ttgtatacac ttattttttt tataacttat ttaataataa aaatcataaa tcataagaaa
2761 ttcgcttatt tagaagtgtc aacaacgtat ctaccaacga tttgaccctt ttccatcttt
2821 tcgtaaattt ctggcaaggt agacaagccg acaaccttga ttggagactt gaccaaacct
2881 ctggcgaaga attgttaatt aagagctcag atcttatcgt cgtcatcctt gtaatccatc
2941 gatactagtg cggccgccct ttagtgaggg ttgaattcga attttcaaaa attcttactt
3001 tttttttgga tggacgcaaa gaagtttaat aatcatatta catggcatta ccaccatata
3061 catatccata tacatatcca tatctaatct tacttatatg ttgtggaaat gtaaagagcc
3121 ccattatctt agcctaaaaa aaccttctct ttggaacttt cagtaatacg cttaactgct
3181 cattgctata ttgaagtacg gattagaagc cgccgagcgg gtgacagccc tccgaaggaa
3241 gactctcctc cgtgcgtcct cgtcttcacc ggtcgcgttc ctgaaacgca gatgtgcctc
3301 gcgccgcact gctccgaaca ataaagattc tacaatacta gcttttatgg ttatgaagag
3361 gaaaaattgg cagtaacctg gccccacaaa ccttcaaatg aacgaatcaa attaacaacc
3421 ataggatgat aatgcgatta gttttttagc cttatttctg gggtaattaa tcagcgaagc
3481 gatgattttt gatctattaa cagatatata aatgcaaaaa ctgcataacc actttaacta
3541 atactttcaa cattttcggt ttgtattact tcttattcaa atgtaataaa agtatcaaca
3601 aaaaattgtt aatatacctc tatactttaa cgtcaaggag aaaaaacccc ggatccgtaa
3661 tacgactcac tatagggccc gggcgtcgac atggaacaga agttgatttc cgaagaagac
3721 ctcgagtaag cttggtaccg cggctagcta agatccgctc taaccgaaaa ggaaggagtt
3781 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg
3841 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt
3901 atactgaaaa ccttgcttga gaaggttttg ggacgctcga agatccagct gcattaatga
3961 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
4021 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
4081 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
4141 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
4201 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
4261 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
4321 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
4381 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
4441 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
4501 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
4561 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
4621 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
4681 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
4741 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
4801 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
4861 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
4921 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
4981 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
5041 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
5101 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
5161 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
5221 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
5281 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
5341 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
5401 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
5461 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
5521 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
5581 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
5641 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
5701 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
5761 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
5821 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
5881 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaacga
5941 agcatctgtg cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac
6001 aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca
6061 acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt
6121 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt
6181 taccaacaaa gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat
6241 ttttctaaca aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc
6301 tcttgataac tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct
6361 attttctctt ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa
6421 gctgcgggtg cattttttca agataaaggc atccccgatt atattctata ccgatgtgga
6481 ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat
6541 tatgaacggt ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc
6601 gtattgtttt cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta
6661 atactagaga taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga
6721 aaggtggatg ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt
6781 ttgagcaatg tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg
6841 cgtttttggt tttttgaaag tgcgtcttca gagcgctttt ggttttcaaa agcgctctga
6901 agttcctata ctttctagag aataggaact tcggaatagg aacttcaaag cgtttccgaa
6961 aacgagcgct tccgaaaatg caacgcgagc tgcgcacata cagctcactg ttcacgtcgc
7021 acctatatct gcgtgttgcc tgtatatata tatacatgag aagaacggca tagtgcgtgt
7081 ttatgcttaa atgcgtactt atatgcgtct atttatgtag gatgaaaggt agtctagtac
7141 ctcctgtgat attatcccat tccatgcggg gtatcgtatg cttccttcag cactaccctt
7201 tagctgttct atatgctgcc actcctcaat tggattagtc tcatccttca atgctatcat
7261 ttcctttgat attggatcat actaagaaac cattattatc atgacattaa cctataaaaa
7321 taggcgtatc acgaggccct ttcgtc
//
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