promoters of S cerevisiae from AEM-2013-Nevoigt

来源: Saccharomyces cerevisiae BY4741
功能: Promoter

Summary

11 mutants of the strong constitutive Saccharomyces cerevisiae TEF1 promoter

基本信息

定量方法 The RNA concentration was quantified by absorbance at 260 nm. Quantification of yECitrine mRNA was performed using the iSciptTM One-Step RT-PCR Kit with SYBR Green (Bio-Rad) according to the manufacturer's instructions and an iCycler (Bio-Rad). We used 100 ng total yeast RNA in one reverse transcription (RT)-PCR.Data were analyzed using the iCycler software (Bio-Rad Laboratories, Hercules, CA)
培养条件 To select and grow yeast transformants using either URA3 or LEU2 as a selectable marker, we used a yeast synthetic complete (YSC) medium containing 6.7 g of Yeast Nitrogen Base (Difco)/liter, 20 g glucose/liter, and a mixture of appropriate nucleotide bases and amino acids (CSM-URA or CSMLEU [Qbiogene, Irvine, CA], respectively), resulting in YSC Leu or YSC Ura . For the growth experiments using a respiratory carbon source, 2% ethanol and 2% glycerol were added to the medium instead of 2% glucose.Yeast cells were routinely cultivated at 30°C in Erlenmeyer flasks closed with cotton plugs and shaken at 200 rpm (semiaerobic conditions) without pH control.Measurements of specific fluorescence were performed using cells harvested from the logarithmic phase during growth in shake flasks.
报告基因 yECitrine
诱导剂
提交人 Yan Xing;Zhao Wenfang

元件序列

元件名 序列 序列长度
Mutant4
ATAGCTTCAAAACGCTTCTACCCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAGACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGCTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGGGACCGCCTCGTTTCTTTCTCTTCGTCGAGAGAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAGAGTTTCTTTCTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGCTAATAAACGGTCTTCAATTTCTCAAGCTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCGTTAGAGAGAAAGCATAGCAATCTAATCTAAGTTC
401
Mutant10
ATAGCTTCAAAATGTTTCTACTCCTTTGTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGAGACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTT-GATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCTTCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTGCTTCTTGTTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTT
401
TEF1p
ATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGAGACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCTTCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTT
401
Mutant7
ATAGCTTCAAAATGTCTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCACCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTCCCCCTCCTCCTTCCTCTAGGGTGCCGTTAATTACCCGTACTAAAGGTTTGGAAAAGGAAAAAGAGACCGCCTCGTCCCTTTTTCTTCGTCGGAGAAGGCAATAAAAATTTTTATCACGTTTCTTTCTCTTGAAAACTTTTTTTTCGATTTTGTTCTCTTTCGACGACCTCCCATTGATATTTGAGTTAACAAACGGTCTTCAATTTCTCAAGTTTCAGCTTCATTTTTCCTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTGGAAAGAAAGCATAGCAATCTAATCTAAGTTT
401
Mutant6
ATAGCTTCAGAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAACTACCCGTACTAAAGGTTTGGAAAA-AAAAAAGAGACCGCCTCGTTTCTTTTTCTTCGTCGCAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCTTCAATTTCTCAAGTTTCAGTTTCATTTTTCTTGCTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTT
401
Mutant3
ATAGCCTCAAAATGTTTCTACTCCTTCTTTACTCTTCCAGATTTTCCCGGACTCCGCGTATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAGAGGAAAAAGAGACCGCCTCGTTTCTCTCTCTTCGTCGGAAAAGGCAATAAAAATTCTTACCACGCTCCTTTTTCTTGAAAATTTTTTTTTTAATTTTTTTCTCTTCCGATGACCTCCCATTGATATTTAGGTTAATAAACGGCCTTTAATTCCTCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATCAGGAAGAAAGCATAGCAATCTAATCTAAGTTT
401
Mutant5
ATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCCTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTCGGAAAAGAAAAAAGAGACCGCCTCGTTTCTTTTTCCTCGCCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCTTCAACTTCTCAAGTCTCAGTCTCATTTTTCTTGTTCTATTACAACTTTTCTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTT
401
Mutant2
ACGGCTCTAAAGTGCTTCGGCTCCCCCTTTACTCCTCCAGGTTTTCTCAGACTCCGCGCATCGCCGTACCACTTCAAAGCCCCCAAGCGCAGCATACCAAATCTCCCCTCTTTCTTCCTCTAGGGTGTCACTAGTTACTCGTACTAAGGGTTTGGGGAAGGAGAAAGAGACCGCCTCGTCTTCTTTTCTTCGTCGAAGGGGGCAATAGAAGTTTTTATCATGTCTCCTTTCCTTGAGAACCTTTTCTTCGATCTTGTTCTCTTTCGACGGCCTCCCGTTGGTATTTAGGTTAATGAACGGTCTTCAACCTCTCAAGTTTCAGTTTCCTTTCTCCCGTCCTATTACGACCCTTCTTACTTCTCACTCAGTAGAACGGGAGCATAGCAATCTAATCCAAGTTT
401
Mutant12
ATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTCTCTCGGACTCCGCACATCGCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAGTTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGACCCAAGCACAGCATACTAAGTTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGAGACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTCTCTTTTTCTTGAAAATTTTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCTTCAATTTCTCCAGTTTCAGTCTCATTTTTCTTGTTCCATTACAACTTTTTTTACTTTTTGCCCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTCT
481
Mutant11
ATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGATTTTCTCGGACTCCGCGCATCGCCGTACCACTTCAAAACGCCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGAGACCGCCTCGTCTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTGATTTTTTTCTCTCTCGATGACCCCCCATTGATATTTAAGTTAATAGACGGTCTTCAATTTCTCAAGTTTCAGTTTCACTTTTCTTGTTCTATTACAACTTTTTTTACTTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTT
401
Mutant8
ATAGCTTCAAAATGTTTCTACTCCTTTTTTACTCTTCCAGACTTTCTCGGACTCCGCGCATCGCCGTACCACTCCCAAACACCCAAGCACAGCATACTAAATTTCCCCTCTTTCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAAAAAAGAGACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAATTTTTATCACGTTTCTTTTTCTTGAAAATTCTTTTTTTGATTTTTTTCTCTTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGCCTTCAATTTCCCAAGTTTCAGTCCCACTTTTCTTGCTCTACTACAACTTTTTTTACCTCTTGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTT
401