| 质粒名称: | pAR-Ec633 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 130873 |
| 存储实验室: | Chang Liu |
|
载体骨架基本信息 |
|
| 载体名称: | CEN/ARS |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 酵母表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | TP-DNAP1 |
| 物种来源: | K. lactis pGKL1/2 system |
| 插入大小: | -- |
| 基因突变: | L477V, L640Y, I777K, W814N |
| GenBank编号 | -- |
| 启动子 | -- |
| 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: | pAR-Ec633 was a gift from Chang Liu (Addgene plasmid # 130873 ; http://n2t.net/addgene:130873 ; RRID:Addgene_130873) |
| 参考文献部分: | Scalable, Continuous Evolution of Genes at Mutation Rates above Genomic Error Thresholds. Ravikumar A, Arzumanyan GA, Obadi MKA, Javanpour AA, Liu CC. Cell. 2018 Dec 13;175(7):1946-1957.e13. doi: 10.1016/j.cell.2018.10.021. Epub 2018 Nov 8. 10.1016/j.cell.2018.10.021 PubMed 30415839 |
买家导航
pAREc633载体基本信息
pAREc633载体图谱质粒图谱和多克隆位点信息
pAREc633载体简介
pAREc633载体序列
LOCUS pAR-Ec633 7602 bp ds-DNA circular SYN 21-AUG-2019
DEFINITION A nuclear plasmid encoding an error-prone mutant TP-DNAP1 (L477V,
L640Y, I777K, W814N) for OrthoRep.
ACCESSION .
VERSION .
KEYWORDS pAR-Ec633
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7602)
AUTHORS Ravikumar A, Arzumanyan GA, Obadi MKA, Javanpour AA, Liu CC
TITLE Scalable, Continuous Evolution of Genes at Mutation Rates above
Genomic Error Thresholds.
JOURNAL Cell. 2018 Dec 13;175(7):1946-1957.e13. doi:
10.1016/j.cell.2018.10.021. Epub 2018 Nov 8.
PUBMED 30415839
REFERENCE 2 (bases 1 to 7602)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 21, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7602
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 271..774
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
primer_bind 815..833
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(871..893)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 993..1012
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 1205..1391
/gene="S. cerevisiae HIS3"
/label=HIS3 promoter
CDS 1392..2054
/codon_start=1
/gene="S. cerevisiae HIS3"
/product="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/label=HIS3
/note="yeast auxotrophic marker"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
terminator 5999..6164
/gene="S. cerevisiae ADH1"
/label=ADH1 terminator
/note="transcription terminator for the S. cerevisiae
alcohol dehydrogenase 1 (ADH1) gene"
primer_bind complement(6054..6078)
/label=pHybLex-R
/note="ADH terminator, reverse primer"
rep_origin complement(6207..6795)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6287..6306)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind complement(7009..7028)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 7020..233
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(7093..7112)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
ORIGIN
1 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
61 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
121 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
181 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
241 tggttaattg gttgtaacac ggtccttttc atcacgtgct ataaaaataa ttataattta
301 aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa ttaaagaaaa
361 aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac aaatactacc
421 ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc ttgtctgtgt
481 agaagaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa tcgaatgtat
541 atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc tttttgttga
601 aattttttaa acctttgttt attttttttt cttcattccg taactcttct accttcttta
661 tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag taaattccca
721 aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc gatccgtcta
781 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
841 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
901 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagtggg
961 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
1021 gcaccatacc accttttcaa ttcatcattt tttttttatt cttttttttg attttaattc
1081 cgttttaaga gcttggtgag cgctaggagt cactgccagg tatcgtttga acacggcatt
1141 agtcagggaa gtcataacac agtcctttcc cgcaattttc tttttctatt actcttggcc
1201 tcctctagta cactctatat ttttttatgc ctcggtaatg attttcattt tttttttcca
1261 cctagcggat gactcttttt ttttcttagc gattggcatt atcacataat gaattataca
1321 ttatataaag taatgtgatt tcttcgaaga atatactaaa aaatgagcag gcaagataaa
1381 cgaaggcaaa gatgacagag cagaaagccc tagtaaagcg tattacaaat gaaaccaaga
1441 ttcagattgc gatctcttta aagggtggtc ccctagcgat agagcactcg atcttcccag
1501 aaaaagaggc agaagcagta gcagaacagg ccacacaatc gcaagtgatt aacgtccaca
1561 caggtatagg gtttctggac catatgatac atgctctggc caagcattcc ggctggtcgc
1621 taatcgttga gtgcattggt gacttacaca tagacgacca tcacaccact gaagactgcg
1681 ggattgctct cggtcaagct tttaaagagg ccctaggggc cgtgcgtgga gtaaaaaggt
1741 ttggatcagg atttgcgcct ttggatgagg cactttccag agcggtggtt gatctttcga
1801 acaggccgta cgcagttgtc gaacttggtt tgcaaaggga gaaagtaggt gatctctctt
1861 gcgagatgat cccgcatttt cttgaaagct ttgcagaggc tagcagaatt accctccacg
1921 ttgattgtct gcgaggcaag aatgatcatc accgtagtga gagtgcgttc aaggctcttg
1981 cggttgccat aagagaagcc acctcgccca atggtaccaa cgatgttccc tccaccaaag
2041 gtgttcttat gtagtgacac cgattattta aagctgctgc atacgatata tatacatgtg
2101 tatatatgta tacctatgaa tgtcagtaag tatgtatacg aacagtatga tactgaagat
2161 gacaaggtaa tgcatcattc tatacgtgtc attctgaacg aggcgcgctt tccttttttc
2221 tttttgcttt ttcttttttt ttctcttgaa ctcgacggat catatatgcg gtgtgaaata
2281 ccgcacagat gcgtaaggag aaaataccgc atcagggagc tcgtgttgtt atccgataca
2341 accggatatt tttcttttaa tgagtctaaa ccgtgatagc ttcaggttaa tacaatcaaa
2401 aaaagctcaa atattctttt aatgccgcgt tcacagattc caattgaata caactaggta
2461 gttcattata tgaagccttt gctactattt ttcactatag tctgccttca ccttaatgca
2521 gacatccaca tattttaatc actttaaaat aaaaaggaag atatattaga agctatgatc
2581 caatctgtaa gccagattaa aattcacgaa ctcttctttc atttgaattg aatgctttga
2641 gttggggtag attatcgcaa attactcatc acatttattg actacgaact tgctgatgtc
2701 ctttttttat ttatattttt cttcagtgaa gcgatttttt ttttacacag accaagacgg
2761 aaaaaagtag ctaaggaaga aaacaaaatc atgaaaaaaa tgtgaagtga tcatgcacat
2821 cgcatcaact taaacattgg cttagagata tatagagtta gagtttacgg caacctttaa
2881 gcaccaatac cttttggcat agtctaaaga cctggttctt aattttaaac aaatttaact
2941 aaagatttcc ctatcaaaga agtaacgagt tgacagattt tctcaaaata aatcgatact
3001 gcatttctag gcatatccag cgagatctat ggactacaaa gacaaagcat taaacgactt
3061 gagaaacgta tacgcagact tcgactcctt gccattggac ttcagacaaa tcttaattaa
3121 ggatagagct acattgttgc aaaaggaaga cgttgaaaag aaaattttgg aaagacaaga
3181 agatgccaaa aagtacgctg aatacttgaa gcaatcagaa atcccagaaa gaatctcctt
3241 acctaacatc aagagacata agggtgtatc aatctccttt gaagaaacat ctgaagatat
3301 ggtcttggaa ccaagacctt ttatcttcga cggtttgaac atcagatgtt tcagaagaga
3361 aaccatattc agtttgaaaa ataagatttt gaacatggtt aaggaatctt catcctttaa
3421 aaacgtttca agacaaagtg tatcttttat gtacttcaaa atttttaata agggtaaagt
3481 tattgcaagt acaaagtctg ttaacatcta tgaagataag atcgacgaaa gattggaaga
3541 tttgtgtaac aacttcgatg acgtattgaa aaagattata gacgtcacct atggttacga
3601 atccttattc gttagtgaaa cttactctta cgtaatattc tatgctaaat caatctactt
3661 tccacaacct agatgcgtta acaactgggg taacaacatc ccaaacatct tgacattcga
3721 ttcctttaag cttttcaccg caaacaaaaa caacgttagt tgtatcaagc aatgttcaag
3781 atttttgtgg caaaaagatt tcaacacttt agaagaaatg atagaataca agaacggtaa
3841 tatttgtatc gtcacaccac aattgcatat caacgatgtt agagacatca agtcattcaa
3901 cgatatcaga ttgtactcag aatcccctat taaaacattt tccgttattg ataacactat
3961 cacatatttg ttttacttca aggaacattt gggtgttatt tttaacatca ctaagtcaag
4021 acatgataga agagtaacaa aattttcacc attgtccaag ttcagtgacg tcaaaaatat
4081 cactgtttgt ttcgatatcg aaagttactt cgacccagaa aaggaatcta accaagttaa
4141 catccctttc atttgttgcg cttcaatcat ctataataag gttattggta acatcgtcga
4201 ttttgaaggt agagactgtg tcgcacaaat gatcgaatac gttgtagata tttgcggtga
4261 attgaacata agttctgttg aattgatcgc ccataacggt ggtggttatg atttccacta
4321 catcttgtca tccatgtata atccagctgc aattaaaaac atattgatca gaaacaactc
4381 ttttatttct tttaatttcg cacatgatgg tgtcaagttc tctgttaagg acagttactc
4441 atttttgttg tgtagtgttg ccaacgcttc taaagcattt ttaaacgaag aaacttttaa
4501 gaaaactgat ttccctcatc acgacttgaa aactgccgat gacttatata aggtctacaa
4561 ggaatggagt tctgttaaca cagaaattaa ccatgtcgtt gaaaaggaaa agttgttgat
4621 cacctctgaa cacatcgtta acttcaccaa gaacgataag agtaagactt tgatcgaatg
4681 gtctaaggat tactgtagaa acgacgtatt ggtcttgtct aaggtttggt tggagtttaa
4741 aaacgctgta gaagatattt ttaactgcga attggtagac caaaccatga ctttggctgg
4801 tttgtcatac aagttgttcc aagcaaacat gccattcgat gtcgaattga gacatcctaa
4861 taaggaagac tacttcaaca tgagagaagc attgattggt ggtagatgta tctctgttaa
4921 cggtatctac aaggatgttt tgtgctatga cgtaaaatca ttatatccag cttctatggc
4981 tttctatgat caaccttacg gttcttttaa gagagtttca tccagaccaa aagatgaatt
5041 gggtatctat tacgttagag taaccccaaa cagaaacaac aagtcaaact ttttccctat
5101 cagatcccat aataagatca cttacaacaa cttcgaagaa tcaacttata ttgcttggta
5161 cacaaacgtt gatatcgaca ttggtttgtc agaaggtcac aacatcgaat acatcccttt
5221 cgattcctat ggtaacattg gttactcatg gtccaaaaag ggtaaaattt ttgaaaagta
5281 catcaaggat gttttgtaca aattgaagat aaaatatgaa aagcaaaata acaaggtaaa
5341 aagaaatgtc atcaaaaaga tcatgaactc attgtggggt aaattcgccc aaaagtgggt
5401 taacttcgaa tacttcatca agtcagaaga tgacattgat ttcgaatccg aagaagcata
5461 taaaattaac gatacagact tcatgttgat caaaaagatt aaggaatcta cctacagttc
5521 taagccaata caaaacggtg tttttacttt aagttgggcc agatatcaca tgaagtctat
5581 ttgggatgca ggtgccaaag aaggtgctga atgtatctat tcagatacag actccatatt
5641 cgttcataag gaacacttca aaaagaatgc aaagtttatg ttgaacggtt tgaaagtacc
5701 aatcatcggt tctgaagtcg gtcaattgga attagaatgt gaatttgata aattgttgtg
5761 cgccggtaaa aagcaataca tgggtttcta cacttacttc caagatggta aaccttgcat
5821 caaggaaaag aaaagattca aaggtatacc aagtaactac atcatacctg aattgtatgc
5881 tcacttgtta tctggtgcag ataaggaagc caaaattcaa tttttaaaat tcagaagaga
5941 atggggttcc gtcaagggtt acatagaaaa taagactgtc aaagccacct aactcgaggc
6001 gaatttctta tgatttatga tttttattat taaataagtt ataaaaaaaa taagtgtata
6061 caaattttaa agtgactctt aggttttaaa acgaaaattc ttattcttga gtaactcttt
6121 cctgtaggtc aggttgcttt ctcaggtata gcatgaggtc gctcactagt ccatggacgc
6181 gtggtaccgg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
6241 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
6301 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
6361 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
6421 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
6481 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
6541 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
6601 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
6661 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
6721 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
6781 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
6841 gaaaactcac gttaagggat tttggtcatg actgatcctt caactcagca aaagttcgat
6901 ttattcaaca aagccacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa
6961 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
7021 tgagccatat tcaacgggaa acgtcttgct ccaggccgcg attaaattcc aacatggatg
7081 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
7141 atcgattgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
7201 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
7261 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
7321 tccccgggaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
7381 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
7441 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
7501 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
7561 aaatgcataa gcttttgcca ttctcaccgg attcagtcgt ca
//
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