| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | PX260 或 pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro |
| 质粒类型: | CRISPR-Cas9载体;基因敲除载体;哺乳动物载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 插入位点: | 限制性内切酶,多克隆位点 |
| 启动子: | U6 |
| 载体大小: | 10203bp |
| 5' 测序引物及序列: | LKO.1 5':GACTATCATATGCTTACCGT |
| 3' 测序引物及序列: | BGH Reverse:TAGAAGGCACAGTCGAGG |
| 载体标签: | 3xFLAG(N ter) |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | Puromycin |
| 克隆菌株: | DH5alpha |
| 宿主细胞(系): | 常规哺乳动物细胞等 |
| 备注: | humanized S. pyogenes Cas9; This plasmid separately encodes a human codon-optimized SpCas9, a tracrRNA and customizable crRNA. |
| 产品目录号: | ZY42229 |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
万千商家帮你免费找货
0 人在求购买到急需产品
- 详细信息
- 文献和实验
- 技术资料
- 保存条件:
-20℃低温保存
- 保质期:
三年
- 英文名:
PX260
- 库存:
20
- 供应商:
泽叶生物
载体基本信息
载体质粒图谱和多克隆位点信息

载体简介
载体序列
LOCUS pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro Sequencing Result 10203 bp DNA SYN
DEFINITION pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro Sequencing Result
ACCESSION
KEYWORDS
SOURCE
ORGANISM other sequences; artificial sequences; vectors.
FEATURES Location/Qualifiers
source 1..10203
/organism="pX260-U6-DR-BB-DR-Cbh-NLS-hSpCas9-NLS-H1-shorttracr-PGK-puro Sequencing Result"
/mol_type="other DNA"
misc_feature complement(168..437)
/label="CAG_enhancer"
misc_feature complement(641..660)
/label="LKO1_5_primer"
rep_origin complement(878..1497)
/label="pBR322_origin"
gene complement(1652..2512)
/label="Ampicillin"
/gene="Ampicillin"
CDS complement(1652..2512)
/label="ORF frame 3"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
promoter complement(2554..2582)
/label="AmpR_promoter"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
misc_feature complement(2741..2763)
/label="pGEX_3_primer"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
rep_origin complement(3031..3337)
/label="f1_origin"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
EIGASLIKHW*"
CDS complement(3538..4485)
/label="ORF frame 1"
/translation="MEAGRPLGQRPIAALLLRFLGSEAGKGWVRGRAQGRAQGRGGRP
KVLRRPGILHASKAHVCRAVLLFLISGPFDLHPSRSRAAEAYHDRVQAHGAPRHPRRR
PQGRTHPRRRVRRLPRHAPHRRSGPPHRAGHRAARTLPHARRARHRQGVGRGRRRRGG
GLDHAGERRSGGGVRRDRPAHGRVERFPAGRAATDGRPPGAAPAQGARVVPGHRRRLA
RPPGQGSGQRRRAPRSGGGRARRGARLPGDLRAPQPPLLRAARLHRHRRRRGARRTAH
LVHDPQARCLTPAPRPAAPDRKERTTPCIDDIRSPGCRN*"
CDS 3576..4253
/label="ORF frame 3"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVVGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLQLLEI*"
gene complement(3627..4226)
/label="puro (variant)"
/gene="puro (variant)"
/translation="MGSCAPFGRALRVVGRASGTGLAGHAPGARSFGHLDVGGDGEAE
PLVEGEVVGRGGLQEGGHPGALGRLHSGEHDGAAQTLALVVGRDADGGQEPRGLLGPV
RRQEAFHLLLRGQPGTAQLGHARADLGEHRPRFDALRRGPDRHRGAVVRDPHLADVEP
DAREEEFLQLGDPLDVAVRIDGVARGGVVGERGGEGAYGPGDVVAGGEAHRGLVLGHG
KLQLLEI*"
CDS complement(3627..4226)
/label="ORF frame 2"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSR
LAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFL
ETSAPHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
misc_feature complement(4314..4333)
/label="mPGK_F_primer"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSR
LAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFL
ETSAPHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
misc_feature 4707..4730
/label="MSCV_rev_primer"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSR
LAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFL
ETSAPHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
misc_feature 4826..4859
/label="loxP"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSR
LAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFL
ETSAPHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
misc_feature complement(4935..4956)
/label="EGFP_C_primer"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSR
LAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFL
ETSAPHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
misc_feature complement(5179..5198)
/label="H1_primer"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSR
LAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFL
ETSAPHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
terminator complement(5354..5557)
/label="bGH_PA_terminator"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSR
LAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFL
ETSAPHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
misc_feature 5543..5560
/label="BGH_rev_primer"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSR
LAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFL
ETSAPHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
CDS complement(5585..9856)
/label="ORF frame 3"
/translation="MDYKDHDGDYKDHDIDYKDDDDKMAPKKKRKVGIHGVPAADKKY
SIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATR
LKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN
IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNS
DVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGL
FGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKN
LSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFD
QSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIP
HQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKS
EETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV
KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVED
RFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLF
DDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDS
LTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENI
VIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQ
NGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEV
VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQ
ILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNA
VVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
风险提示:丁香通仅作为第三方平台,为商家信息发布提供平台空间。用户咨询产品时请注意保护个人信息及财产安全,合理判断,谨慎选购商品,商家和用户对交易行为负责。对于医疗器械类产品,请先查证核实企业经营资质和医疗器械产品注册证情况。
文献和实验手把手教你利用 CRISPR-Cas9 系统精准敲除靶标基因
2 Cas9 - 2 hitKO 系统三、Cas9 - 2 hitKO 系统中涉及到的载体为了达到高效的敲除效率,我们在同一个载体重引入 cas9 酶和两个向导性 RNA。整个系统包括 3 个载体: PX458M ,PX459M 和 EZ-GuideXH。PX458M ,PX459M 除了含有 Cas9 和一个向导性 RNA 插入位点,还引入第二个向导性 RNA 插入位点。EZ-GuideXH 是为插入第二个向导性 RNA 的辅助性载体。PX458M 带有 EGFP 荧光标记; PX459M 带有
CRISPR/Cas9 是一种强大的基因组编辑工具,今天我们介绍 CRISPR/Cas9 系统操作。以下操作说明是基于作者冰糖个人实验室操作,仅供参考。 质粒介绍我们使用的是 Feng Zhang 实验室的系统的 pX330 质粒,如下图所示: 酶切系统 37 ℃, 3 hr;Run 1% gel,回收。Elute 到 50 mL H2O。我们利用 BbsI 消化载体形成粘末端,以利于下游 oligo 退火产物的连接;酶切时请充分,且绝对不能加 CIP 处理
1. Qubit 定量与紫外分光光度计定量的区别 在常规实验室对核酸定量的检测中,常会用到紫外分光光度计(或 Nanodrop)和 Qubit 两种检测方法,二者在原理、准确性、操作难易等方面存在显著差异。 ①首先两者原理不同,紫外分光光度计基于核酸的紫外吸收特性来实现对核酸的定量:核酸(DNA/RNA)在 260nm 波长处有特异性吸收峰,且吸光度与核酸浓度成正比;Qubit 基于荧光染料与核酸特异性结合的原理来实现对核酸的定量,两者结合后荧光强度与核酸浓度成正比,通过标准曲线校准实现定量
技术资料暂无技术资料 索取技术资料




