The prokaryotic promoter gb2 is a slightly modified version of the Em7 promoter. It mediates higher transcription efficiency than the normally used Tn5 promoter. The promoter of the mouse Phosphoglucokinase gene (PGK) is used as eukaryotic promoter. A synthetic polyadenylation signal terminates the kanamycin/neomycin expression. Vector with a minimal promoter and an NFAT response element for studying cell signaling using destabilized luciferase. This promoter fragment was obtained by PCR using primers PGK‐FZ and PGK‐RZ (Table 1), which were designed to introduce a BstBI site immediately upstream of the ATG translation initiation codon and a BglII site at the 5′ end of the P PGK1 sequence, respectively. The resulting plasmid was named pPGKAMYB. For this promoter, there is a cluster of 2 transcription start sites,in the interval 3072928 to 3072929, where the highest frequency is at position 3070951. The most upstream position of this cluster is assigned as TSS position in the promoter. After blast, this promoter sequence is equal to the "3109bp……3628bp" in GenBank: KT351864.1. But I found another PGK promoter in a patent, which sequence is equal to the "127bp……518bp" in Gene Bank...
The regulatory sequences of the Pgk-1 gene were used to drive the E. coli lacZ reporter gene and 2 strains of transgenic animals created with this Pgk-lacZ transgene carried on autosomes . Using the P19 embryonal carcinoma cells as recipients, we found that constructs carrying the promoter and intragenic regions of the murine Pgk-1 gene were ... GenBank File: Plasmid sequence and annotations. Use text editor or plasmid mapping software to view sequence. ... pLEX-MCS PGK promoter Sequences (1)
Liver toxicity is the major concern for use of recombinant human tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) proteins in treatment of cancers. Here we report that normal human primary hepatocytes (NHPHs) are susceptible to the transduction of the wild-type, full-length coding sequence of the human TRAIL gene. To minimize potential toxicity of the TRAIL gene, a bicistronic ... Our 3rd generation lentiviral systems have been designed for increased researcher safety. pLenti-CAG-GFP-PGK-RFP-T2A-Puro Lentiviral Reporter Plasmid contains GFP reporter driven by CAG promoter, RFP and Puromycin resistance gene are driven by PGK promoter respectively. This dual reporter vector serves a positive control. (Promoter modules are composite regulatory elements consisting of at least two transcription factor binding sites separated by a spacer. Promoter modules can have synergistic or antagonistic functions (1).) Note: The synthetic Ampr in the pGL4 Vectors expresses a protein sequence identical to that expressed by the Ampr gene in the pGL3 Vectors. This sequence cooperates with DMPK promoter elements, one of which binds Sp1, to up-regulate transcription in myoblasts. These data, in conjunction with other studies showing MyoD responsiveness of the DMPK transcript ( 15 , 16 , 58 ), implicate this myogenic factor in regulating transcription of this gene in muscle. The analysis of transformed strains in which single copies of lacZ fusions have been targeted to the qutE locus has identified three constitutive positively acting sequence elements in the pgk gene. Firstly, sequence located between -161 and -120bp (possibly spanning -120) relative to the transcript start site is essential for expression, thus demonstrating that the putative core promoter, including potential TATA and CCAAT boxes, and a pyrimidine rich region, is not alone sufficient to ...
This sequence cooperates with DMPK promoter elements, one of which binds Sp1, to up-regulate transcription in myoblasts. These data, in conjunction with other studies showing MyoD responsiveness of the DMPK transcript ( 15 , 16 , 58 ), implicate this myogenic factor in regulating transcription of this gene in muscle. Using internal deletions and insertions we have identified the elements in the PGK promoter which are required for correct RNA initiation at the CAAG sequence at -39. The results indicate that two different components of the PGK promoter contribute to correct RNA initiation, the TATA homologies, located at -152 and -113, and the sequences at the site of initiation. Plasmid pPGKpuro from Dr. Rudolf Jaenisch's lab contains the insert pgk-1 5' and 3' regions and is published in Genes Dev. 1996 Apr 15. 10(8):1008-20. This plasmid is available through Addgene.
Jun 18, 2018 · The presently disclosed subject matter provides compositions and methods for the expression of CRISPR guide RNAs using the H1 promoter. In particular, compositions and methods are provided for the use of the H1 promoter to express CRISPR guide RNA (gRNA) with altered specificity of the 5′ nucleotide, as well as use of the H1 promoter sequence as a bidirectional promoter to express Cas9 ... InvivoGen provides pDRIVE plasmids bearing the HSP70 promoter that drives the expression of a reporter gene (SEAP or LacZ). HSP70 is the promoter of Heat Shock Protein 70 with a specificity: Ubiquitous, heat inducible. pDRIVE (LacZ) and pDRIVE5-SEAP plasmids are selectable in E.coli and in mammalian
May 12, 2010 · Surprisingly, with the exception of SV40, all the mammalian promoters produced above-background levels of GFP signals, with PGK producing a level of expression that approaches that of COPIA and ACT5C . It is interesting that PGK, which is a weak promoter in mammalian cells, can behave as a fairly strong promoter in fly cells.
Jun 09, 2018 · Its 5' to 3' sequence is the same as in the database (ACCESSION AE000141). (b) Presented so that the promoter sequence numbers increase from left to right. Its 5' to 3' sequence is comparable to the consensus sequence. 16. lac promoter- lactose- (or IPTG- isopropylthiogalactoside) inducible Problems with lac promoter- 1. Weak 2. Promoters are about 100-1000 base pairs long and are adjacent and typically upstream (5') of the sense or coding strand of the transcribed gene. The coding strand is the DNA strand that encodes codons and whose sequence corresponds to the mRNA transcript produced.
The results revealed that the 1.4 kb DNA region is sufficient for determining the organ‐specific, developmental stage‐specific and spermatogenic stage‐specific transcription of the mouse Pgk‐2 gene. When the region between – 684 and + 61 was used to generate transgenic mice, β‐galactosidase mRNA was detectable not only in the testis, but also in other organs such as brain and lung. Its strength has been estimated in K. marxianus by putting the homologous β-glucosidase gene under its control. This promoter is very efficient as activities higher than the ones obtained with the Saccharomyces cerevisiae PGK promoter were obtained. This promoter appears to be constitutive in various conditions tested.
Plasmid pPGKpuro from Dr. Rudolf Jaenisch's lab contains the insert pgk-1 5' and 3' regions and is published in Genes Dev. 1996 Apr 15. 10(8):1008-20. This plasmid is available through Addgene.
compares favorably with the SV40 promoter. Although CpG islands are characteristically unmethylated (Bird 1986), and the PGK-1 island on the Xa is unmethylated, the Xi is methylated at 119 of 121 CpG dinucleotides analyzed in this region (Pfeifer et al. 1990a, b). Our earlier Jun 18, 2018 · The presently disclosed subject matter provides compositions and methods for the expression of CRISPR guide RNAs using the H1 promoter. In particular, compositions and methods are provided for the use of the H1 promoter to express CRISPR guide RNA (gRNA) with altered specificity of the 5′ nucleotide, as well as use of the H1 promoter sequence as a bidirectional promoter to express Cas9 ... It combines a prokaryotic promoter (gb2) for expression of kanamycin resistance in E.coli with a eukaryotic promoter (PGK) for expression of neomycin resistance in mammalian cells. The included 706-Cre expression plasmid allows Cre recombination in E.coli. This kit can also be used to work on bacterial artificial chromosomes.
Plasmid PGK-H2BmCherry from Dr. Mark Mercola's lab contains the insert Phosphoglycerate kinase promoter and is published in PLoS ONE. 2009 . 4(4):e5046. This plasmid is available through Addgene.The prokaryotic promoter gb2 is a slightly modified version of the Em7 promoter. It mediates higher transcription efficiency than the normally used Tn5 promoter. The promoter of the mouse phosphoglucokinase gene (PGK) is used as eukaryotic promoter. A synthetic polyadenylation signal terminates the kanamycin/neomycin expression.
In some embodiments, the truncated PGK promoter comprises or consists of a nucleic acid sequence of a WT PGK promoter (e.g., SEQ ID NO: 1) with a partial deletion in the WT sequence (e.g., a deletion of at least 20, 25, 50, 100, 150, 200, 250, 300, 350, 400, or 450 nucleic acids).
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Phosphoglycerate kinase (PGK) is a key enzyme involved in the metabolism of glucose via glycolysis. In most mammals, PGK is encoded by two genes, Pgk1 and Pgk2 (VandeBerg et al. 1973, VandeBerg 1985, McCarrey & Thomas 1987).
Hi All, I want to put my EGFP under the PGK promoter of Puromycin through IRES for mammalian syst... Removing all stop codons from Sequence Record using Biopython Hello Everyone, I have sequences that occasionally have an erronious stop codon. sequences are perfectly matched to the miR-142 sequence. In the present report, we show that addition of 4 copies of ... (PGK) promoter were previously described.8 Constitutive promoters are used routinely to drive ectopic gene expression. Here, we carried out a systematic comparison of eight commonly used constitutive promoters (SV40, CMV, UBC, EF1A, PGK and CAGG for mammalian systems, and COPIA and ACT5C for Drosophila systems). We also included in the comparison the TRE promoter, which can be activated by the rtTA transcriptional activator in a ...