Certificate of Analysis Takara Bio USA, Inc.1290 Terra Bella Avenue, Mountain View, CA 94043, USA U.S. Technical Support: ********************United States/Canada 800.662.2566 Asia Pacific+1.650.919.7300Europe+33.(0)1.3904.6880Japan+81.(0)77.565.6999Page 1 of 6Tet-On® 3G Vector Set (with ZsGreen1)Table of ContentsDescription (1)pCMV-Tet3G Vector Information (2)pTRE3G-ZsGreen1 Vector and pTRE3G-Luc Control Vector Information (4)Quality Control Data (6)Catalog No. Lot Number631159 (Not sold separately) Specified on product label.DescriptionThe Tet-On 3G Vector Set (with ZsGreen1) is used to create tightly regulated and highly responsive tetracycline (Tet)-inducible mammalian expression systems that are turned on by the addition of doxycycline to the culture medium. The Tet-On 3G Vector Set (with ZsGreen1) allows the simultaneous expression of a gene of interest and a green fluorescent protein marker.Package Contents•20 μl pCMV-Tet3G Vector (500 ng/μl)•20 μl pTRE3G-ZsGreen1 Vector(500 ng/μl)•20 μl pTRE3G-Luc Control Vector(500 ng/μl)•40 μl Linear Hygromycin Marker (50 ng/μl)•40 μl Linear Puromycin Marker (50 ng/μl)Storage Conditions•Store plasmids at –20°C.•Spin briefly to recover contents.•Avoid repeated freeze/thaw cycles.Shelf Life• 1 year from date of receipt under proper storage conditions.Storage Buffer•10 mM Tris-HCl (pH 8.0), 1 mM EDTA (pH 8.0)Shipping Conditions• Dry ice (–70°C)Product DocumentsDocuments for our products are available for download at /manualsThe following documents apply to this product:•Tet-On 3G Expression Systems User Manual (PT5148-1)pCMV-Tet3G Vector InformationFigure 1. pCMV-Tet3G Vector Map.DescriptionThe pCMV-Tet3G Vector expresses Tet-On 3G, a tetracycline-controlled transactivator that exhibits high activity in the presence of the inducer doxycycline (Dox), and exceptionally low activity in its absence. Tet-On 3G results from the fusion of amino acids 1–207 of a mutant Tet repressor (TetR) to 39 amino acids that form three minimal "F"-type transcriptional activation domains from the herpes simplex virus VP16 protein. Tet-On 3G was derived from Tet-On Advanced (Zhou et al. 2006; Urlinger et al. 2000; Gossen and Bujard 1992; Gossen et al. 1995); as a result, it’s fully synthetic, lacks cryptic splice sites, and is codon-optimized for stable expression in mammalian cells. Compared to both of its predecessors, however, this 3rd generation Tet-On transactivator demonstrates increased sensitivity to Dox (Zhou et al. 2006). Constitutive expression of Tet-On 3G is driven by the human cytomegalovirus immediately early promoter (P CMV IE).Location of Features in pCMV-Tet3G•P CMV IE(human cytomegalovirus immediate early promoter): 2–688•Tet-On 3G (transactivator gene): 775–1521•SV40 polyA signal: 1536–1991•pUC origin of replication: 2342–2996•Amp r (ampicillin resistance gene; β-lactamase): 3144–4004 (complementary)•SV40 polyA signal: 4275–4809 (complementary)•Kan r/Neo r (kanamycin/neomycin resistance gene): 5417–6211 (complementary)•P SV40 e (SV40 early promoter): 6532–6891 (complementary)Additional InformationpCMV-Tet3G is used to develop stable Tet-On 3G cell lines, which are hosts for Tet-inducible gene expression systems. To create a Tet-inducible expression system, a vector containing a gene of interest under the control of the Tet-inducible TRE3G promoter(P TRE3G) is transfected into a Tet-On 3G cell line. The addition of Dox to the system causes Tet-On 3G to undergo a conformational change that allows it to bind to P TRE3G, activating transcription of the gene of interest in a highly dose-dependent manner. Additional information on TRE-containing vectors, and protocols describing the construction of Tet-On 3G cell lines can be found in the Tet-On 3G Expression Systems User Manual (PT5148-1).Propagation in E. coli•Suitable host strain: Stellar™ Competent Cells•Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.• E. coli replication origin: pUCpTRE3G-ZsGreen1 Vector and pTRE3G-Luc Control Vector InformationFigure 2. pTRE3G-ZsGreen1 Vector and pTRE3G-Luc Control Vector Maps.Figure 3. pTRE3G-ZsGreen Vector Multiple Cloning Site. The internal start site (ATG) at the IRES2/MCS junction is indicated in bold.DescriptionpTRE3G-ZsGreen1is a Tet-inducible, mammalian expression vector designed to coexpress a gene of interest and the green fluorescent protein ZsGreen1 under the control of the Tet-responsive promoter P TRE3G. This promoter consists of a highly optimized Tet-responsive element (TRE) just upstream of a minimal CMV promoter. P TRE3G exhibits exceptionally low basal activity; it’s induced by the binding of Tet-On 3G but is virtually silent in its absence. The vector is designed to be used as part of our Tet-On 3G Inducible Expression System (Cat. No. 631164).ZsGreen1 is a human codon-optimized variant of the reef coral Zoanthus sp. green fluorescent protein (ZsGreen) that has been engineered for brighter fluorescence (excitation and emission maxima: 493 and 505 nm, respectively; Matz et al. 1999; Haas, Park, and Seed 1996). p TRE3G-ZsGreen allows Dox-inducible coexpression of ZsGreen1 and a gene of interest from a bicistronic mRNA transcript. An encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES2), positioned between ZsGreen1 and the gene of interest, facilitates cap-independent translation of the gene of interest from an internal start site at the IRES2/MCS junction (Jang et al. 1988). This ensures that a high percentage of ZsGreen1-expressing clones also express the gene of interest, allowing ZsGreen1 to be used as an indicator of inducibility and transfection efficiency, as well as a marker for selection by flow cytometry. The vector also contains a pUC origin of replication and an ampicillin resistance gene (Amp r) to allow for propagation and selection in E. coli.The pTRE3G-Luc is a Tet-inducible control vector that expresses firefly luciferase under the control of P TRE3G. When used with standard luciferase detection reagents, this vector can be used as a reporter of induction efficiency (see User Manual for protocol). pTRE3G-Luc is not intended to be used as a cloning vector.Location of Features in pTRE3G-ZsGreen1•P TRE3G (3rd generation Tet-responsive promoter): 7–382•ZsGreen1: 389–1084•IRES2 (encephalomyocarditis virus internal ribosome entry site): 1091–1673•MCS (multiple cloning site): 1686–1721•SV40 polyA signal: 1776–2573•pUC origin of replication: 2838–3481•Amp r (ampicillin resistance gene; β-lactamase): 3629–4489 (complementary)Location of Features in pTRE3G-Luc•P TRE3G (3rd generation Tet-responsive promoter): 7–382•Luciferase: 432–2084•SV40 polyA signal: 2151–2948•pUC origin of replication: 3213–3856•Amp r (ampicillin resistance gene; β-lactamase): 4004–4864 (complementary)Additional InformationpTRE3G-ZsGreen1 is a mammalian expression vector that allows tightly regulated, doxycycline-controlled coexpression of a gene of interest and ZsGreen1. The gene of interest must have both a start and a stop codon. The gene of interest should be cloned in-frame with the start codon at the IRES2/MCS junction (this codon is shown in bold in the MCS sequence in Figure 3, page 3; see the User Manual for details on how to use In-Fusion® to simplify your cloning). Cotransfection of pTRE3G-ZsGreen1 constructs with Linear Hygromycin or Puromycin Markers allows antibiotic selection of stable transfectants. In order to function, the system requires the presence of the Tet-On 3G transactivator protein, supplied by a stable Tet-On 3G cell line created with our Tet-On 3G Inducible Expression System (Cat. No. 631164).Propagation in E. coli•Suitable host strain: Stellar™ Competent Cells•Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.• E. coli replication origin: pUCExcitation and Emission of pTRE3G-ZsGreen1•Excitation: 493 nm•Emission: 505 nmReferences•Gossen, M. et al. Transcriptional activation by tetracyclines in mammalian cells. Science268, 1766–9 (1995).•Gossen, M. & Bujard, H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. U. S. A.89, 5547–51 (1992).•Haas, J., Park, E. C. & Seed, B. Codon usage limitation in the expression of HIV-1 envelope glycoprotein. Curr.Biol.6, 315–24 (1996).•Jang, S. K. et al. A segment of the 5’ nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation. J. Virol.62, 2636–43 (1988).•Matz, M. V et al. Fluorescent proteins from nonbioluminescent Anthozoa species. Nat. Biotechnol.17, 969–73 (1999).•Urlinger, S. et al. Exploring the sequence space for tetracycline-dependent transcriptional activators: novel mutations yield expanded range and sensitivity. Proc. Natl. Acad. Sci. U. S. A.97, 7963–8 (2000).•Zhou, X., Vink, M., Klaver, B., Berkhout, B. & Das, A. T. Optimization of the Tet-On system for regulated gene expression through viral evolution. Gene Ther.13, 1382–90 (2006).Quality Control DataPlasmid Identity & Purity•Digestion with the indicated restriction enzymes produced fragments of the indicated sizes on a 0.8% agarose/EtBr gel:Vector Enzyme(s) Fragment(s)pCMV-Tet3G EcoRI 7.1 kbEcoRI & HindIII 1.2 & 5.9 kbpTRE3G-ZsGreen1 XhoI 4.7 kbEcoRV 1.2 & 3.5 kbpTRE3G-Luc XhoI 5.1 kbEcoRI & BamHI 2.1 & 3.0 kbLinear Hygromycin Marker HindIII & XbaI0.5, 0.6 & 1.1 kbLinear Puromycin Marker HindIII & XbaI0.45, 0.6, & 0.75 kb•Vector identity was confirmed by sequencing.•A260/A280: 1.8–2.0Functional Testing of Linear Markers•HEK 293 cells were transfected with 200 ng of either the Linear Hygromycin Marker or the Linear Puromycin Marker. After 5 hr at 37°C, the transfection solution was removed, and the cells were given fresh medium. 48 hr later, the cells were plated in two 10 cm plates. 48 hr after plating, medium containing either hygromycin orpuromycin (depending on the linear marker used to transfect the cells) was added to the plates. After 2–3 weeks, >20 clones were identified.It is certified that this product meets the above specifications, as reviewed and approved by the Quality Department.CATALOG NO.631159NOTICE TO PURCHASER:Our products are to be used for research purposes only. They may not be used for any other purpose, including, but not limited to, use in drugs, in vitro diagnostic purposes, therapeutics, or in humans. Our products may not betransferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without prior written approval of Takara Bio USA, Inc.Your use of this product is also subject to compliance with the licensing requirements listed below and described on the product´s web page at . It is your responsibility to review, understand and adhere to any restrictions imposed by these statements.STATEMENT 24The RCFPs (including DsRedExpress, DsRedExpress2, and E2-Crimson) are covered by one or more of thefollowing U.S. Patent Nos. 7,166,444; 7,157,565; 7,217,789; 7,338,784; 7,338,783; 7,537,915; 6,969,597; 7,150,979;7,442,522 and 8,012,682.STATEMENT 72Living Colors Fluorescent Protein Products: Not-For-Profit Entities: Orders may be placed in the normal manner by contacting your local representative or Takara Bio USA, Inc. Customer Service. Any and all uses of this product will be subject to the terms and conditions of the Non-Commercial Use License Agreement (the “Non-Commercial License”), a copy of which can be found below. As a condition of sale of this product to you, and prior to using this product, you must agree to the terms and conditions of the Non-Commercial License. Under the Non-Commercial License, Takara Bio USA, Inc. grants Not-For-Profit Entities a non-exclusive, non-transferable, non-sublicensable and limited license to use this product for internal, non-commercial scientific research use only. Such licensespecifically excludes the right to sell or otherwise transfer this product, its components or derivatives thereof to third parties. No modifications to the product may be made without express written permission from Takara Bio USA, Inc.Any other use of this product requires a different license from Takara Bio USA, Inc. For license information, please ***************************************************************************************.For-Profit Entities wishing to use this product are required to obtain a license from Takara Bio USA, Inc. For license information, please contact a licensing representative by phone at 650.919.7320 or by e-mail at ***********************.STATEMENT 42Use of the Tetracycline controllable expression systems (the "Tet Technology") is covered by a series of patents including U.S. Patent # 7541446, # 8383364, # 9181556 , European patents EP # 1200607, # 1954811, #2352833Academic research institutions are granted an automatic license with the purchase of this product to use the Tet Technology only for internal, academic research purposes, which license specifically excludes the right to sell, or otherwise transfer, the Tet Technology or its component parts to third parties. Notwithstanding the above, academicand not-for profit research institutions whose research using the Tet Technology is sponsored by for profitorganizations, which shall receive ownership to any data and results stemming from the sponsored research, shall need a commercial license agreement from TET Systems in order to use the Tet Technology. In accepting this license, all users acknowledge that the Tet Technology is experimental in nature. TET Systems GmbH & Co. KG makes no warranties, express or implied or of any kind, and hereby disclaims any warranties, representations, or guarantees of any kind as to the Tet Technology, patents, or products. All others are invited to request a license from TET Systems GmbH & Co. KG prior to purchasing these reagents or using them for any purpose. Takara Bio USA, Inc. is required by its licensing agreement to submit a report of all purchasers of the Tet-controllable expression system to TET Systems.For license information, please contact:GSF/CEOTET Systems GmbH & Co. KG,Im Neuenheimer Feld 58269120 Heidelberg GermanyTel: +49 6221 5880400Fax: +49 6221 5880404email:*******************or use the electronic licensing request form via /ip-licensing/licensing/for-profit-research TRADEMARKS:© 2015 Takara Bio Inc. All Rights Reserved.All trademarks are the property of Takara Bio Inc. or its affiliate(s) in the U.S. and/or other countries or their respective owners. Certain trademarks may not be registered in all jurisdictions.。
TBT交叉偶极声波处理TBT是斯伦贝谢公司过钻头测井系统ThruBits的新型声波测井仪器，有SonicScanner小型化之称，结构示意图如下图1。
主要特点一是无割缝仪器外壳设计，各模式井筒波响应及频散效应可数学模拟[1]，二是CHIRP偶极声波发射器具有连续、宽频、高信噪比等优势，三是12级独立的接收器组较大程度地改善了波形数据质量。
其接收器间距为4英寸，单极发射器到第1组接收器间距为5.85英尺、偶极发射器到第1组接收器间距为6.5英尺。
图1 Thrubits TBT仪器结构示意图目前Thrubit交叉偶极声波处理软件已完成了数据预处理，纵波、横波、斯通利波慢度及能量提取等功能模块的开发测试。
针对页岩气水平井TBT测井数据特征形成了余弦镶边带通滤波技术、波形可视化刻度技术，全波慢度、能量提取技术。
软件时差能量分析模块已投入生产使用，在川渝页岩气23井次的TBT交叉偶极声波处理中取得了较好的应用效果，各向异性分析模块正在改进完善中。
一、TBT关键采集信息及数据预处理斯伦贝谢公司过钻头测井系统ThruBits提供给用户的野外数据格式为DLIS 文件，其中包括了单极发射记录的12组高频全波波形和12组低频全波波形，它们分别用于纵波和斯通利波信息提取，如下表1所示。
偶极发射记录4 个方位共48组横波波形，主要用于地层横波信息提取及各向异性分析，如下表2所示。
表1 Thrubits TBT单极方式关键采集信息表2 Thrubits TBT交叉偶极方式关键采集信息测井时，接收器除了接收到我们需要的有用声波以外，还接收了沿井眼或井壁传播的其他类型波，它们的存在会对计算结果产生较大影响。
在对波形信息提取前，需要对各组波形开展带通滤波处理工作，特别是针对小型化的过钻头交叉偶极声波资料，对各组波形进行带通滤波处理非常重要。
带通滤波处理是以快速傅立叶变换理论为基础，针对水平井页岩气测井资料，我们设计了余弦镶边带通滤波器，取得了较好的应用效果，其频率响应如图2所示。
dGB Earth Sciences. 专注于地球物理和地学软件解决方案OpendTect简介F3 –荷兰海上工区演示数据P.F.M. de Groot博士dGB Earth Sciences B.V.Nijverheidstraat 11-27511 JM EnschedeThe NetherlandsTel.; +31 53 4315155Fax,;+31 53 4315104e-mail; info@dgb-groupwebsite; dgb-group吴新天中国北京市海淀区花园东路32号花园公寓A1111目录1 关于OpendTectOpendTect是一个开源环境下的地震解释系统。
用户可以利用OpendTect的属性及现代可视化技术，如视频显示和体透视等三维可视化技术进行多个地震数据体的处理、浏览和解释。
还可通过一些免费和商业化的插件拓展OpendTect的功能。
1.1 OpendTect Base系统1.1.1 可视化地震解释员要求能够快速地浏览多个数据体，将所获得的信息综合起来以最优地理解研究对象的地质特征。
因此，OpendTect综合了先进的数据处理和可视化功能。
可视化对象可以灵活地在各数据空间中移动，以交互分析存储的数据或实时计算的数据。
体透视和视频浏览的功能是为了更好地理解数据和成果。
1.1.2 地震属性在OpendTect中，地震属性用于对地震数据进行筛选以及目标检测。
OpendTect在设计之初就旨在为解释人员提供最大的数据透视性，而且属性结果能够通过OpendTect的交互工作流程得到优化。
1.1.3 层位解释OpendTect提供多种层位追踪和解释选项。
同时还提供层位编辑以及层位计算等多种功能。
1.2 插件更多具体工作可借助商业插件dGB 以及第三方软件商为OpendTect 提供的插件来完成。
1.2.1 倾角控制倾角控制插件允许用户创建并使用控制数据体，这种控制数据体在每一个样点位置上都提取地震同相轴的倾角和方位角信息。
