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GenBank File: Plasmid sequence and annotations. Use text editor or plasmid mapping software to view sequence. ... pLEX-MCS PGK promoter Sequences (1) Phosphoglycerate kinase (PGK, E.C. 2.7.2.3) interconverts ADP + 1,3-bisphospho-glycerate (1,3-bPGA) to ATP + 3-phosphoglycerate (3PGA). While most bacteria have a single pgk gene and mammals possess two copies, plant genomes contain three or more PGK genes. In this study, we identified five Pgk genes in the Zea mays var. B73 genome, predicted to encode proteins targeted to different ...

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...

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Here we used postnatal day 1 systemic injection of self-complementary adeno-associated virus (scAAV9) vectors carrying a codon-optimized SMN1 sequence and a chimeric intron placed downstream of the strong phosphoglycerate kinase (PGK) promoter (SMNopti) to overexpress the human SMN protein in a mouse model of severe SMA. May 30, 2010 · The MBP promoter did not support eGFP expression in neurons (not shown), but strongly directed eGFP expression in cultured oligodendocytes. A version of PGK-GFP containing Lck sequence fused with GFP (PGK–Lck-GFP) also efficiently transduced neurons; the Lck sequence resulted in expression localized to the cell surface and neurites.

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)

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contain the endogenous PGK upstream activation sequence located, in the natural PGK promoter, between nucleotide C Scoordinates -423 and -470.Scar 2. A hybrid yeast promoter according to claim 1, in which the GAL10 upstream activation sequence is e provided at a site from which the PGK upstream activation sequence has been deleted. 3. The upstream activator sequence is contained in the 5' region of the yeast PGK gene between nucleotides -324 and -455. The upstream activator sequence has synthetic linkers attached to either end to facilitate attachment to a vector. Vectors including the upstream activator sequence and a heterologous promoter sequence are described.

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 ...

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A role for Cpflp at the PGK promoter was examined using a cpfl null strain of yeast. Northern blot analysis was used to assay transcription from the chromosomal PGK gene in the absence of Cpf1p, and also transcription from a multicopy plasmid carrying the wild type PGK gene in the cpf1- background. May 30, 2010 · The MBP promoter did not support eGFP expression in neurons (not shown), but strongly directed eGFP expression in cultured oligodendocytes. A version of PGK-GFP containing Lck sequence fused with GFP (PGK–Lck-GFP) also efficiently transduced neurons; the Lck sequence resulted in expression localized to the cell surface and neurites.

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.

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We also show Yersinia pestis, Pseudomonas aeruginosa, Shewanella that fbaA is cotranscribed with pgk, from promoter putrefaciens and Vibrio cholerae, the pgk gene is followed epd P0 or an internal pgk P1 promoter of the extended by the fbaA gene encoding a class II FBA and instead of -10 class. Minimal Promoter-Driven NanoLuc® Reporter Vectors. Two vectors that offer a minimal promoter for cloning response elements of interest to express NanoLuc® luciferase. N1031, N1041. Promoter-Driven Control NanoLuc® Luciferase Vectors. Three NanoLuc® reporter vectors with CMV, PGK or TK promoter.

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

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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). I introduced the CAG promoter from the pCXN2 vector to the PacI site of the STOP-EFGP-ROSA TV, between SA and the floxed Neo-STOP cassette. 5. R-A4 From the Stuart Orkin lab at the Children’s Hospital, Boston. This is not a targeting vector. Instead, it is a plasmid containing a 4kb genomic DNA from the Rosa 26 locus.

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.

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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. The respective transcriptional initiation sites in front of gap and pgk were located, and, from the analysis of DNA sequences upstream of these and of previously determined transcriptional start sites, common structures which may be important for promoter function in C. glutamicum are discussed.

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.

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The human ubiquitin C sequences described here allow expression of transgenes in an even wider range of tissues compared to the promoter/enhancer units that are routinely used. Transgenes driven by H2-K b , CMV or Pgk-1 regulatory sequences are highly expressed in spleen, lung and salivary glands (H2-K b ; 1 ), in spleen, stomach and heart (CMV ...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

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.

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The PGK promoter has been inserted between the ClaI and BamHI sites and contains sites downstream to allow a second gene to be inserted. The BsgI and BseRI restriction sites in the first MCS cleave within the stop codon in the XbaI site and allow the retrospective fusion of coding sequences. Sequence Subsequent pop-out of the promoter-URA3 would result in only one copy of the desired promoter replacing the HIS3 promoter . In order to simplify the experimental procedure, we designed upstream and downstream primers each containing a 39-base sequence identical to the target chromosome locus followed by a 20-base sequence identical to the ...

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.

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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 The Pgk-1 promoter is in a region rich in nucleotides G and C. This promoter can efficiently drive high levels of expression of reporter genes such as E. coli lacZ and neo. We have determined that the 120 bp upstream of the transcription start site functions as a core promoter.

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.

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Pgk-1 is the murine gene encoding phosphoglycerate kinase (PGK), a key enzyme in glycolysis. This enzyme must be present in all cells, so the Pgk-1 promoter is expected to be a ubiquitously active one. However, the Pgk-1 promoter is regulated. The Pgk-1 gene resides on the X chromosome and becomes transcriptionally silenced when X chromosomes become inactivated in female somatic cells or in ...The pMS1 vector also has an internal ribosome entry site (IRES) to enable reporter gene detection of trapped cell surface and secreted proteins as well as to allow reporter gene detection of trapping events that would otherwise not maintain the correct open reading frame. pGTlox2, 3 and 4 vectors are engineered to utilize the recombinase ...

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.

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• CMV promoter : nt234- 802 • Multiple cloning sites: nt880 -946 • PGK promoter transcription cassette: nt1159-3409 • WPRE sequence: 3420- 4008 Note: 1. Since lentiviral transfer vector may undergo sequence re -arrangement and/or deletion when its ligated products were transformed into DH5 α competent Our analysis revealed that PGK has an upstream activation sequence (UASPGK) located between 402 and 479 nucleotides upstream from the initiating ATG sequence which is required for full transcriptional activity. Deletion of this sequence caused a marked reduction in the levels of PGK transcription.

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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The use of PGK promoter derived from S. cerevisiae in place of the E. coli-derived promotor raised the APH expression efficiency to a level equivalent to that of an auxotrophic marker (Hadfield et al., Curr. Genet., 18, 303(1990)). Here we used postnatal day 1 systemic injection of self-complementary adeno-associated virus (scAAV9) vectors carrying a codon-optimized SMN1 sequence and a chimeric intron placed downstream of the strong phosphoglycerate kinase (PGK) promoter (SMNopti) to overexpress the human SMN protein in a mouse model of severe SMA.

A repository of over 200,000 plasmids including Protein Structure Initiative protein expression plasmids and vectors, over 75,000 human plasmids, and whole genome collections from many organisms.

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Summary of PGK1 expression in human tissue. Ubiquitous cytoplasmic and nuclear expression.PGK-DTa Probe NheI Hind III Hind III 11.5 kb Hind III PGK-DTa 8.2 kb Site III Site IV Probe SphI SphI 12 kb Probe Bgl II Bgl II SphI SphI PGK-DTa Bgl II Fig. S1 Probe EcoRI PGK Neo GFP PGK Puro VMC PGK Puro K14-Agouti oeN KGP Puro

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).

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Aug 11, 2011 · Plasmid construction. The promoter sequences for chicken β-actin , human polypeptide chain elongation factor 1α (EF1α) , mouse phosphoglycerate kinase (PGK) , human ubiquitin C (UbC) , polyoma enhancer/herpes simplex virus thymidine kinase (MC1) , immediate early enhancer of human cytomegalovirus (CMV), a deletion derivative of CMV (CMVd1) and CMV immediate early enhancer/chicken β-actin ... XX RN [2] RC plasmid from pUC19 & linkers RC pFL34 from plasmid & URA3 gene RC pFL40 from pFL34 & yeast 2 micron ori RC pFL60 from pFL40 & PGK promoter/terminator & linker RA Minet M., Lacroute F.; RT "Cloning and sequencing of a human cDNA coding for a multifunctional RT polypeptide of the purine pathway by complementation of the ade2-101 RT ...

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 ...