| 质粒名称: | pYES2.1-TRP1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 11007 |
| 存储实验室: | Francisco Malagon |
|
载体骨架基本信息 |
|
| 载体名称: | pYES2.1/V5-His-TOPO |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 5886 bp |
| 完整质粒大小: | 6558 bp |
| 载体类型: | 细菌表达，酵母表达 |
| 筛选标记: | URA3 |
| 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: | TRP1 |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 672 bp |
| 基因突变: | ORF-TRP1 without stop codon |
| 标签/融合蛋白: |
his6 (C terminal on backbone)
V5 (C terminal on backbone)
|
| 克隆方法: | TOPO Cloning |
| 5' 测序引物 | T7 |
|
文献引用方法 |
|
| 材料和方法部分: | pYES2.1-TRP1 was a gift from Francisco Malagon (爱迪基因 plasmid # 11007) |
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 pYES2.1-TRP1 6558 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pYES2.1-TRP1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6558)
TITLE Unpublished, Paco
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pYES2.1-TRP1|
FEATURES Location/Qualifiers
source 1..6558
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 2..443
/gene="S. cerevisiae GAL1"
/label=GAL1 promoter
/note="inducible promoter, regulated by Gal4"
protein_bind 2..119
/label=UAS
/bound_moiety="Gal4"
/note="upstream activating sequence mediating
Gal4-dependent induction"
primer_bind 414..437
/label=GAL1
/note="S. cerevisiae GAL1 promoter, forward primer"
primer_bind 475..494
/label=T7
/note="T7 promoter, forward primer"
promoter 475..493
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 513..1184
/codon_start=1
/gene="S. cerevisiae TRP1"
/product="phosphoribosylanthranilate isomerase, required
for tryptophan biosynthesis"
/label=TRP1
/note="yeast auxotrophic marker"
/translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
KK"
CDS 1215..1256
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 1266..1283
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
terminator 1304..1551
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(1320..1338)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
primer_bind complement(1628..1645)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1799..2387)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1879..1898)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2558..3418)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCDTLLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLID*MEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKRSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
CDS complement(3514..4317)
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
promoter complement(4318..4533)
/gene="S. cerevisiae URA3"
/label=URA3 promoter
rep_origin 5137..6017
/label=2u ori
/note="yeast 2u plasmid origin of replication"
rep_origin complement(6086..6541)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 6435..6454
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 acggattaga agccgccgag cgggtgacag ccctccgaag gaagactctc ctccgtgcgt
61 cctcgtcttc accggtcgcg ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga
121 acaataaaga ttctacaata ctagctttta tggttatgaa gaggaaaaat tggcagtaac
181 ctggccccac aaaccttcaa atgaacgaat caaattaaca accataggat gataatgcga
241 ttagtttttt agccttattt ctggggtaat taatcagcga agcgatgatt tttgatctat
301 taacagatat ataaatgcaa aaactgcata accactttaa ctaatacttt caacattttc
361 ggtttgtatt acttcttatt caaatgtaat aaaagtatca acaaaaaatt gttaatatac
421 ctctatactt taacgtcaag gagaaaaaac cccggatcgg actactagca gctgtaatac
481 gactcactat agggaatatt aagctcgccc ttatgtctgt tattaatttc acaggtagtt
541 ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga ggccgcagaa tgtgctctag
601 attccgatgc tgacttgctg ggtattatat gtgtgcccaa tagaaagaga acaattgacc
661 cggttattgc aaggaaaatt tcaagtcttg taaaagcata taaaaatagt tcaggcactc
721 cgaaatactt ggttggcgtg tttcgtaatc aacctaagga ggatgttttg gctctggtca
781 atgattacgg cattgatatc gtccaactgc atggagatga gtcgtggcaa gaataccaag
841 agttcctcgg tttgccagtt attaaaagac tcgtatttcc aaaagactgc aacatactac
901 tcagtgcagc ttcacagaaa cctcattcgt ttattccctt gtttgattca gaagcaggtg
961 ggacaggtga acttttggat tggaactcga tttctgactg ggttggaagg caagagagcc
1021 ccgaaagctt acattttatg ttagctggtg gactgacgcc agaaaatgtt ggtgatgcgc
1081 ttagattaaa tggcgttatt ggtgttgatg taagcggagg tgtggagaca aatggtgtaa
1141 aagactctaa caaaatagca aatttcgtca aaaatgctaa gaaaaagggc gagcttcgag
1201 gtcacccatt cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta
1261 ccggtcatca tcaccatcac cattgagttt ctagagggcc gcatcatgta attagttatg
1321 tcacgcttac attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga
1381 caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta
1441 tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata
1501 ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg ctttaatttg caagctgcgg
1561 ccctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
1621 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
1681 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
1741 tgtgagcaaa aggccagcaa aagcccagga accgtaaaaa ggccgcgttg ctggcgtttt
1801 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
1861 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
1921 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
1981 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
2041 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
2101 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
2161 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
2221 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
2281 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
2341 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
2401 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
2461 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
2521 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
2581 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
2641 agataactac gatacgggag cgcttaccat ctggccccag tgctgcaatg ataccgcgag
2701 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
2761 gcagaagtgg tcctgcaact ttatccgcct ccattcagtc tattaattgt tgccgggaag
2821 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttggcatt gctacaggca
2881 tcgtggtgtc actctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
2941 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
3001 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
3061 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
3121 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
3181 ataatagtgt atcacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
3241 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
3301 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
3361 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
3421 tcttcctttt tcaatgggta ataactgata taattaaatt gaagctctaa tttgtgagtt
3481 tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa
3541 atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt
3601 tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca catcatccac
3661 ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat
3721 aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat
3781 aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga
3841 tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac
3901 ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt
3961 cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt
4021 attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa
4081 tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt
4141 ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt
4201 acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt
4261 ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat
4321 ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt
4381 tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct
4441 tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaagaa accgaaatca
4501 aaaaaaagaa taaaaaaaaa atgatgaatt gaattgaaaa gctagcttat cgatgataag
4561 ctgtcaaaga tgagaattaa ttccacggac tatagactat actagatact ccgtctactg
4621 tacgatacac ttccgctcag gtccttgtcc tttaacgagg ccttaccact cttttgttac
4681 tctattgatc cagctcagca aaggcagtgt gatctaagat tctatcttcg cgatgtagta
4741 aaactagcta gaccgagaaa gagactagaa atgcaaaagg cacttctaca atggctgcca
4801 tcattattat ccgatgtgac gctgcagctt ctcaatgata ttcgaatacg ctttgaggag
4861 atacagccta atatccgaca aactgtttta cagatttacg atcgtacttg ttacccatca
4921 ttgaattttg aacatccgaa cctgggagtt ttccctgaaa cagatagtat atttgaacct
4981 gtataataat atatagtcta gcgctttacg gaagacaatg tatgtatttc ggttcctgga
5041 gaaactattg catctattgc ataggtaatc ttgcacgtcg catccccggt tcattttctg
5101 cgtttccatc ttgcacttca atagcatatc tttgttaacg aagcatctgt gcttcatttt
5161 gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
5221 tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca
5281 tttttgtaaa acaaaaatgc aacgcgacga gagcgctaat ttttcaaaca aagaatctga
5341 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
5401 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
5461 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
5521 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
5581 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
5641 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
5701 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
5761 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
5821 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
5881 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
5941 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
6001 gaagcggtat tcgcaatggg aagctccacc ccggttgata atcagaaaag ccccaaaaac
6061 aggaagattg tataagcaaa tatttaaatt gtaaacgtta atattttgtt aaaattcgcg
6121 ttaaattttt gttaaatcag ctcatttttt aacgaatagc ccgaaatcgg caaaatccct
6181 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttc caacaagagt
6241 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaagggtcta tcagggcgat
6301 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
6361 gtaaatcgga agggtaaacg gatgccccca tttagagctt gacggggaaa gccggcgaac
6421 gtggcgagaa aggaagggaa gaaagcgaaa ggagcggggg ctagggcggt gggaagtgta
6481 ggggtcacgc tgggcgtaac caccacaccc gccgcgctta atggggcgct acagggcgcg
6541 tggggatgat ccactagt
//
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